Immunopotentiator, composition for immunostimulation, and TNF - α production promoter

Abstract

To provide a gout ameliorating agent and a gout ameliorating composition having excellent gout ameliorating action and high safety, and to provide a liver function improving agent and a liver function improving composition having excellent immunopotentiating action. To provide an immunopotentiator and a composition for immunopotentiation having high safety, an anti-aging agent and a composition for anti-aging having excellent anti-aging action and high safety, an anti-inflammatory agent and a composition for anti-inflammation having excellent anti-inflammatory action and high safety, a hair grower and a composition for hair growth having excellent hair growing action and high safety, and an antioxidant and a composition for antioxidant having excellent antioxidant action and high safety.SOLUTION: The gout improver, the liver function improver, the immunopotentiator, the anti-aging agent, the anti-inflammatory agent, the hair grower or the antioxidant contains an extract of Peucedanum japonicum Thunb.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to gout-improving agents and compositions for gout improvement, liver function-improving agents and compositions for liver function improvement, immunostimulants and compositions for immunostimulation, anti-aging agents and compositions for anti-aging, anti-inflammatory agents and compositions for anti-inflammatory use, hair growth agents and compositions for hair growth, and antioxidants and compositions for antioxidant use. [Background technology]

[0002] Xanthine oxidase is an enzyme that catalyzes the conversion of hypoxanthine to xanthine and then to uric acid in nucleic acid metabolism. In recent years, dietary habits have changed rapidly, and the number of people consuming high-calorie, high-protein, and high-fat diets has increased, resulting in an annual increase in the number of gout patients. Gout, known as a lifestyle-related disease, is caused by hyperuricemia due to abnormal purine metabolism, and is a disease that causes severe pain in joints such as the big toe. This pain occurs when increased uric acid in the blood crystallizes and deposits in the joints. Therefore, controlling blood uric acid levels within normal limits is considered the basis for preventing and treating diseases such as gout and hyperuricemia.

[0003] To date, drugs for adjusting blood uric acid levels, such as the uric acid synthesis inhibitor "allopurinol," have been provided. However, allopurinol has problems such as its transient xanthine oxidase inhibitory activity and the accompanying side effects. Furthermore, a xanthine oxidase inhibitor containing a resveratrol derivative as an active ingredient has also been proposed (see, for example, Patent Document 1).

[0004] Furthermore, due to their inhibitory activity against the generation of reactive oxygen species, xanthine oxidase inhibitors are expected to be useful in the treatment of diseases involving reactive oxygen species, for example, in the treatment of cardiovascular diseases through the improvement of vascular function (see, for example, Non-Patent Document 1).

[0005] The liver is an extremely important organ for humans, performing many vital functions, such as metabolizing and storing various nutrients absorbed by the intestine, as well as producing and secreting bile, detoxifying and excreting. However, the liver is susceptible to acute or chronic damage caused by various factors, including irregular lifestyles, stress, viruses, drugs, alcohol, malnutrition, and hepatic circulatory disorders, which can lead to diseases (liver dysfunction) such as acute hepatitis, chronic hepatitis, fatty liver, jaundice, and cirrhosis.

[0006] Glutathione is a tripeptide composed of three amino acids, glutamic acid, cysteine, and glycine, and is a compound containing the major intracellular cysteine ​​residue. In the liver, glutathione not only protects hepatocytes from various oxidative stresses but also directly contributes to the expression of liver functions such as drug metabolism by forming conjugates with toxic substances such as drugs and reactive compounds and excreting them extracellularly. However, the exertion of glutathione's effects also results in the consumption of glutathione. Indeed, it is known that liver damage induced in rats with galactosamine or other substances reduces the amount of glutathione in hepatocytes. Therefore, promoting glutathione production in the liver is thought to suppress liver damage and ultimately improve liver function. Hot water extracts of amber (see, for example, Patent Document 2) are known to have the effect of promoting glutathione production.

[0007] Furthermore, in order to promote cell proliferation, it is important to supply cells with the energy necessary for cell division. ATP is an example of an energy substance in living organisms, and it is thought that increasing the amount of ATP produced promotes intracellular energy metabolism, leading to cell proliferation. However, it has been reported that the amount of ATP, an energy substance, is reduced in cells with reduced function or aging cells compared to normal cells (see, for example, Patent Document 3). Therefore, if ATP production in cells can be promoted, it is thought that the cells can be activated, cell division can be promoted, and the proliferation ability of the cells can be restored.

[0008] In recent years, consumers have become increasingly health conscious. However, modern society is flooded with factors that damage the immune system, such as irregular lifestyles, unbalanced diets, and mental stress. It is known that a weakened immune system can lead to various diseases, including cancer, infectious diseases, and allergic symptoms. Conversely, stimulating the immune system can be expected to have a variety of effects, including carcinogenesis suppression, anti-cancer effects, anti-infectious diseases, anti-allergic effects, and even the restoration of physical rhythms and the maintenance of homeostasis.

[0009] Many types of cells are involved in the immune system, but white blood cells play a particularly important role. Macrophages are present universally in all animals and are an important type of white blood cell involved in all stages of the immune response, including the early stages. In recent years, the function of white blood cells has been elucidated at the material level, and it has become clear that the functions of white blood cells and intercellular interactions are carried out by trace proteins (cytokines) secreted by white blood cells.

[0010] There are many types of cytokines, among which tumor necrosis factor-α (TNF-α) and interleukins (ILs) have attracted attention. Among these, inflammatory cytokines, typified by TNF-α, are mainly released from macrophages and have been reported to ultimately exhibit antitumor effects. Therefore, it is believed that enhancing the production of TNF-α can suppress the growth of malignant tumors. Based on this idea, extracts from plants belonging to the genus Ribes in the Saxifragaceae family (see, for example, Patent Document 4) have been proposed as substances that promote TNF-α production.

[0011] Skin is composed of the stratum corneum, epidermis, basement membrane, and dermis. The basement membrane is located at the boundary between the epidermis and dermis and not only connects the epidermis and dermis but also plays an important role in maintaining skin function (see, for example, Non-Patent Document 2). The basement membrane's main skeleton has a mesh structure made of type IV collagen. Various glycoproteins, primarily composed of laminin-332 (also known as laminin-5), are located at the boundary between the basement membrane and epidermis and connect the two. Laminin-332 is produced by epidermal keratinocytes present in the epidermis. In young skin, the interaction between the epidermis and dermis is maintained at homeostasis through the function of the basement membrane, ensuring moisture retention, flexibility, elasticity, etc., and maintaining the skin's appearance of firmness, luster, and freshness.

[0012] The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as collagen, elastin, hyaluronic acid, etc., which are present outside these cells and support the skin structure. In young skin, fibroblast proliferation is active, and the interaction between fibroblasts, extracellular matrix components, and other skin tissues maintains homeostasis, ensuring moisture retention, flexibility, elasticity, etc., and maintaining the skin in a firm, glossy, and moist state in appearance.

[0013] However, when exposed to certain external factors such as ultraviolet radiation, extremely dry air, or excessive skin washing, or when aging occurs, laminin-332, a major component of the basement membrane, degrades and alters, destroying the basement membrane structure (see, for example, Non-Patent Document 3). Furthermore, the production of collagen, elastin, hyaluronic acid, and other major components of the extracellular matrix decreases, leading to their degradation and alteration.

[0014] As a result, the skin's moisturizing function and elasticity decline, and abnormal peeling of the keratin occurs, causing the skin to lose its firmness and luster, leading to aging symptoms such as rough skin and wrinkles. These changes associated with skin aging, such as wrinkles, dullness, loss of texture, and loss of elasticity, are related to the decrease in basement membrane components, structural changes in the basement membrane, and the decrease and denaturation of extracellular matrix components such as collagen, elastin, and hyaluronic acid.

[0015] Therefore, promoting the production of laminin, collagen, and hyaluronic acid is important for preventing, treating, or improving skin aging.

[0016] Laminins consist of various combinations of α, β, and γ chains, and 17 types are currently known. These α, β, and γ chains form a coiled triple-stranded structure at the long arm, forming a large laminin molecule. Among these, laminin-332, consisting of α3, β3, and γ2 chains, is abundantly present in the basement membrane of epithelial tissues such as the skin, digestive tract, kidney, and lung. A genetic disease (lethal congenital epidermolysis bullosa, or Herlitz junctional epidermolysis bullosa) caused by congenital abnormalities in the genes encoding each chain of laminin-332 is known to cause fatal symptoms, including complete epidermal peeling. Compared to other extracellular matrix molecules, laminin-332 is known to strongly adhere cells (high cell adhesion activity) and strongly promote cell motility (high cell motility activity) (see, for example, Non-Patent Document 4).

[0017] As described above, laminin-332 is known to promote cell migration in damaged skin and promote wound healing due to its high cell motility activity (see, for example, Patent Document 5). In other words, promoting the production of laminin-332 is important for promoting the healing of skin injuries that destroy the basement membrane structure.

[0018] Previously, topical skin preparations containing laminin-332 have been known (see, for example, Patent Document 6), and other substances known to promote laminin-332 production include soybean extract (see, for example, Patent Document 7), phenylpropanoids (see, for example, Patent Document 8), pantothenic acid (see, for example, Patent Document 9), lysophosphatidylcholine or lysophosphatidic acid (see, for example, Patent Document 10), and coenzyme Q10 (see, for example, Patent Document 11).

[0019] Among the above-mentioned extracellular matrix components, type VII collagen is a major component of anchoring fibers that connect the epidermal basement membrane and dermis. Type VII collagen is biosynthesized by epidermal keratinocytes and dermal fibroblasts, transported extracellularly, and then forms anchoring fibers. Anchoring fibers are thought to strengthen the connection between the epidermal basement membrane and dermis and play an important role in maintaining the structure of the skin.

[0020] Here, type VII collagen, which constitutes anchoring fibers, is known to decrease with both physiological aging and photoaging (see, for example, Non-Patent Document 5), and the accompanying structural changes in the skin are thought to be one of the causes of the appearance of signs of aging such as wrinkles and sagging, and the accompanying decline in skin function. Therefore, if the production of type VII collagen can be promoted, it is thought that it will be possible to prevent and improve skin aging symptoms such as wrinkles and sagging, and the accompanying decline in skin function.

[0021] Furthermore, when strong external forces or friction are applied to the skin, wounds that damage the skin structure itself or blisters that destroy the bond between the dermis and epidermis occur. Repair and stabilization of the basement membrane are essential for healing these wounds, and promoting the production of type VII collagen is thought to be useful in treating such wounds and blisters.

[0022] Conventionally, substances known to have type VII collagen production promoting activity include, for example, a solvent extract of yuzu seeds (see, for example, Patent Document 12) and an okra extract (see, for example, Patent Document 13).

[0023] Among the above-mentioned extracellular matrix components, hyaluronic acid is a type of mucopolysaccharide that fills the intercellular spaces to hold cells in place, and also has numerous other functions, such as retaining moisture in the intercellular spaces, providing lubrication and flexibility to tissues, and resisting external forces such as mechanical damage. Promoting hyaluronic acid production is believed to prevent, treat, or improve skin aging symptoms such as rough skin, wrinkles, dullness, changes in texture, loss of elasticity, and loss of moisturizing function. Furthermore, promoting the expression of hyaluronan synthase 3 (HAS3), which is involved in promoting the synthesis of epidermal hyaluronic acid, is believed to prevent, treat, or improve skin aging.

[0024] Hyaluronic acid is also present in connective tissues such as cartilage, synovial fluid, the umbilical cord, the vitreous body, and other tissues, in addition to skin tissue. Hyaluronic acid in synovial fluid coats the surface of articular cartilage, contributing to the smooth functioning of joints through its lubricating and protective functions. Meanwhile, the concentration of hyaluronic acid in synovial fluid is known to decrease in cases of arthritis, such as rheumatoid arthritis. Therefore, promoting hyaluronic acid production may be effective in preventing or treating arthritis, including rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, and osteoarthritis. Furthermore, granulation tissue forms during wound or burn healing, and hyaluronic acid levels are known to increase significantly in granulation tissue. Therefore, promoting hyaluronic acid production may promote wound or burn healing. Licorice leaf extract (see, for example, Patent Document 14) and the like are known to have the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA.

[0025] The epidermis functions to mitigate external stimuli and control the loss of body components such as water, and is composed of a four-layer structure, starting with the basal layer at the bottom, followed by the spinous layer, granular layer, and stratum corneum. Most of the cells in each layer are keratinocytes that differentiate from the basal layer. Keratinocytes that divide and proliferate in the basal layer differentiate as they pass through the spinous and granular layers to become corneocytes, which then form the stratum corneum, made up of keratin protein fibers with strong cross-links, and are eventually shed from the stratum corneum as dirt.

[0026] Among the stratum basale, stratum spinosum, stratum granulosum, and stratum corneum that make up the epidermis, the cell membrane thickens to form a thickened cell membrane, and the action of transglutaminase-1 (TGM-1) causes glutamyl-lysine crosslinking between protein molecules to form strong keratin protein fibers. Furthermore, ceramides and other molecules covalently bond to some of these fibers, forming a hydrophobic structure that provides a foundation for the lamellar structure of intercellular lipids and forms the basis for the keratin barrier function.

[0027] However, as the amount of transglutaminase-1 produced in the epidermis decreases with age, the stratum corneum barrier function and the skin's moisturizing function decline, resulting in the onset of skin aging symptoms such as rough skin and dry skin, and dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.). Therefore, it is believed that promoting the production of transglutaminase-1 in the epidermis can prevent, treat, or improve skin aging symptoms and dry skin diseases. Extracts from Hunan sweet tea (see, for example, Patent Document 15) are known to have the effect of promoting transglutaminase-1 production.

[0028] The stratum corneum is the outermost layer of the skin and serves as a physical barrier against external stimuli. To maintain this barrier function, the skin undergoes a cycle of epidermal metabolism (keratinization), typically repeated every four weeks, from the production of keratinocytes in the basal layer until they become dandruff and are shed. However, the metabolic function of the stratum corneum also declines with age, resulting in skin problems such as fine wrinkles, dullness, pigmentation, and rough skin. Therefore, promoting keratinocyte proliferation and restoring skin's metabolic function is thought to improve skin aging symptoms such as fine wrinkles, dullness, and pigmentation. Extracts of Aster crustacea (see, for example, Patent Document 16) and the like are known to have the effect of promoting epidermal keratinocyte proliferation.

[0029] In skin cells, aquaporins, known as water channels, are expressed on the cell membrane and are known to take up small molecules such as water from the intercellular space into the cells. Thirteen types of aquaporins (AQP0-AQP12) are known to exist in humans. Epidermal cells primarily contain AQP3, which is thought to take up not only water but also small molecules such as glycerol and urea, which are involved in moisture retention.

[0030] However, AQP3 decreases with age, and it has been suggested that this is one of the reasons for the decline in water retention function. Therefore, it is thought that promoting AQP3 expression may be able to control age-related changes in water retention ability and barrier function (see, for example, Non-Patent Document 6). For example, extracts from star fruit leaves (see, for example, Patent Document 17) are known to have the effect of promoting AQP3 expression.

[0031] Filaggrin is a component of the skin and is thought to be involved in the barrier function of the skin, preventing the invasion of allergens, toxins, and infectious organisms. It is known that a decrease in filaggrin function due to gene mutations or the like is associated with the risk of developing atopic diseases, including atopic dermatitis (eczema, skin inflammation, skin itching, etc.), allergies, asthma, etc., and in more severe cases, can lead to skin diseases such as ichthyosis vulgaris (see, for example, Non-Patent Document 7).

[0032] Meanwhile, amino acids, the main components of natural moisturizing factors (NMFs), are produced by the degradation of filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilaggrin in epidermal keratinocytes present in the granular layer just below the stratum corneum. It is then immediately phosphorylated, accumulates in keratohyalin granules, and is degraded to filaggrin through dephosphorylation and hydrolysis. It then migrates to the stratum corneum, where it increases the aggregation efficiency of keratin filaments and is known to be involved in the internal organization of keratinocytes (see, for example, Non-Patent Document 8). In recent years, it has been discovered that filaggrin is extremely important and essential for skin moisture retention, and that conditions such as dryness reduce the ability of filaggrin synthesis, resulting in a decrease in the amount of amino acids in the stratum corneum (see, for example, Non-Patent Document 9).

[0033] Therefore, promoting filaggrin production in epidermal keratinocytes is thought to be able to prevent, treat, or improve atopic diseases, including atopic dermatitis (eczema, skin inflammation, skin itching, etc.), allergies, asthma, etc. Furthermore, promoting filaggrin production and thereby increasing the amount of amino acids in the stratum corneum is expected to essentially improve the moisture environment of the stratum corneum.

[0034] Furthermore, it was previously believed that the skin's barrier function was solely the responsibility of the stratum corneum. However, genetic deletion of the constituent proteins of tight junctions (hereinafter sometimes abbreviated as TJs) present in the granular layer of the epidermis disrupts the skin's barrier function. Therefore, in recent years, TJs have been considered to also play an important role in the skin's barrier function (see, for example, Non-Patent Document 10). TJs are junctions that not only bring adjacent cells into close contact with each other, but also control the permeation of substances by sealing the gaps between cells. Proteins that make up TJs include claudins (CLDN), occludin (OCLN), ZO-1, and ZO-2. These proteins are thought to form the framework of TJ strands and control the barrier function of TJs (see, for example, Non-Patent Document 11). Based on the above, if the expression of claudins, occludin, ZO-1, or ZO-2 is reduced for some reason, structural destruction of TJs occurs, and they no longer function as a permeation barrier to substances, which is expected to cause skin symptoms such as dry skin, rough skin, atopic dermatitis, and various infectious diseases.

[0035] Therefore, it is believed that promoting the production of claudins, occludin, ZO-1, and ZO-2 in the epidermis, thereby promoting TJ formation in epidermal keratinocytes, can enhance the skin's barrier function and moisture retention function, thereby preventing or ameliorating the above-mentioned skin symptoms. Based on this idea, natural products such as Coptis japonica extract (see, for example, Patent Document 18) and Picea abies extract (see, for example, Patent Document 19) have been disclosed as substances that improve skin barrier function through the action of promoting TJ formation.

[0036] Promoting ATP production in skin cells is important for promoting skin turnover, restoring the skin's metabolic function, and preventing and improving skin aging such as wrinkles, dullness, loss of texture, etc. Glycogen (see, for example, Patent Document 3 mentioned above) and extracts from natural products such as turmeric (see, for example, Patent Document 20) are known to have the effect of promoting ATP production.

[0037] Glutathione in cells functions as a radical scavenger, cellular function regulator through redox, xenobiotic metabolism, and sulfhydryl donor for various enzymes, and is also known as an antioxidant against reactive oxygen species. Its activity is thought to be derived from the cysteine ​​residue. However, it has been reported that glutathione levels in cells become deficient or decrease due to excessive oxidative stress, the addition of xenobiotics, aging, etc. This is thought to be one of the factors that reduces the cell's defense against oxidative stress and damages components such as DNA and proteins in cells.

[0038] Diseases known to be pathologically associated with a decrease or deficiency in intracellular glutathione levels include pigmentation such as skin blemishes, skin aging, and the aforementioned diseases caused by oxidative stress, as well as liver damage (caused by excessive alcohol consumption or the ingestion of foreign substances such as heavy metals and chemicals). In other words, promoting glutathione production is thought to enhance the cellular defense against oxidative stress and prevent or treat the aforementioned diseases caused by a decrease or deficiency in intracellular glutathione levels. Hot water extracts of amber (see, for example, Patent Document 2, cited above) are known to have the effect of promoting glutathione production.

[0039] The Maillard reaction is a reaction in which the amino groups of amino acids, peptides, and proteins react with ketones, aldehydes, and especially reducing sugars such as glucose to produce brown pigments. The substances produced as the end products of this reaction are called advanced glycation end products (AGEs). The Maillard reaction consists of an early reaction in which amino groups react non-enzymatically with glucose to form Schiff bases, followed by an Amadori rearrangement. The middle reaction further produces activated intermediates with dicarbonyl groups, such as 3-deoxyglucosone (3-DG). The activated intermediates further react non-enzymatically with amino groups, undergoing repeated dehydration and condensation reactions to form AGEs.

[0040] Identified AGEs include imidazolone (see, for example, Non-Patent Document 12), Nε-carboxymethyllysine (CML) (see, for example, Non-Patent Document 13), pentosidine, pyrraline, crosslin, Nε-carboxyethyllysine, methylglyoxallysine dimer, glyoxallysine dimer, etc. Imidazolone has been reported to be produced by the reaction of 3-DG with arginine (see, for example, the above-mentioned Non-Patent Document 12).

[0041] One of the pathologies in which AGEs are involved in the onset and progression is aging symptoms. The progression of the Maillard reaction in biological tissues leads to aging (decreased elasticity) in skin tissue due to cross-linking of skin elastic fibers, and the deposition of AGEs in blood vessel wall tissue and neurofibrils is also said to lead to arteriosclerosis and Alzheimer's disease.

[0042] As a natural product having an AGE formation inhibitory effect, for example, an extract of the pericarp of Diarium indum, a legume, has been disclosed (see, for example, Patent Document 21). In addition, compounds such as aminoguanidine, OPB-9195, and pyridoxamine are known to have the effect of inhibiting the formation of AGEs, but these compounds have problems such as side effects (see, for example, Non-Patent Documents 12 to 14).

[0043] Inflammatory diseases, such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, and other inflammatory skin diseases accompanied by rough skin, as well as rheumatoid arthritis, osteoarthritis, asthma, etc., have diverse causes and onset mechanisms. Known causes include tumor necrosis factor (TNF-α), which is mainly produced by macrophages, increased hyaluronidase activity, histamine release, and increased prostaglandin (PG) E2 production (enhanced cyclooxygenase-2 (COX-2) activity).

[0044] TNF-α was discovered as a factor that induces tumor necrosis, but it has recently been considered to be a cytokine that not only acts against tumors but also plays a mediator role in regulating the function of normal cells. TNF-α plays an important role in the process of inflammation from its onset to its resolution, but its sustained and excessive production can cause damage to tissues, including the skin, and can cause fever and cachexia systemically, leading to the worsening of inflammation. Therefore, in pathological inflammation, it is important to suppress excessive TNF-α production. Extracts from Earthworm Moth (see, for example, Patent Document 16 mentioned above) are known to have the effect of suppressing TNF-α production.

[0045] Histamine release is a phenomenon in which histamine in mast cells is released outside the cells, and the released histamine causes an inflammatory reaction. Therefore, attempts have been made to prevent or treat allergic diseases and inflammatory diseases using substances that inhibit or suppress histamine release. However, it is difficult to directly evaluate histamine release, and histamine release can be evaluated using the release of hexosaminidase, which has been confirmed to be released simultaneously with histamine release, as an indicator. Therefore, by inhibiting the release of hexosaminidase, histamine release can also be inhibited, which is thought to be effective in preventing, treating, or improving inflammatory diseases, etc.

[0046] Histamine also mediates intercellular communication as a local neurotransmitter, promoting gastric acid secretion in the digestive tract and functioning as a neurotransmitter in the central nervous system, contributing to maintaining wakefulness. Excessive histamine release can cause ulcers due to gastric hyperacidity in the digestive tract and contributes to sleep disorders in the central nervous system. As mentioned above, inhibiting the release of hexosaminidase can also inhibit the release of histamine, which is thought to prevent, treat, or improve gastric ulcers, sleep disorders, and other conditions caused by gastric hyperacidity. For example, extracts from wisteria tea (see, for example, Patent Document 22) are known to have the effect of inhibiting hexosaminidase release.

[0047] Inflammation is a complex reaction that manifests with symptoms such as redness, edema, fever, pain, and functional impairment. Microscopically, inflammation consists of common reactions, such as vascular responses resulting in plasma leakage, leukocyte infiltration, and tissue destruction by inflammatory cells. It can also cause systemic reactions, including fever and hyperalgesia, which involve the central nervous system. Prostaglandins play an important role in each of these inflammatory reactions, and it has been shown that the production of prostaglandins during inflammation is primarily mediated by cyclooxygenase-2, an inducible cyclooxygenase. For this reason, many cyclooxygenase inhibitors, such as aspirin, are used to prevent and prevent inflammatory reactions (see, for example, Non-Patent Document 15).

[0048] Hair grows and falls out repeatedly according to a cyclical hair cycle consisting of an anagen phase, a catagen phase, and a telogen phase. The stage of this hair cycle, from telogen to anagen, during which new hair follicles are formed, is considered to be the most important for hair growth, and dermal papilla cells are thought to play an important role in the proliferation and differentiation of hair follicle epithelial cells during this stage. Dermal papilla cells are located inside hair follicle epithelial cells, which are composed of outer root sheath cells and matrix cells near the hair root, in the basement membrane-enclosed shaft portion of the hair root. They act on hair follicle epithelial cells to promote their proliferation, and thus play an important role in the proliferation and differentiation of hair follicle epithelial cells and hair formation (see, for example, Non-Patent Document 16).

[0049] Thus, dermal papilla cells play an important role in the proliferation and differentiation of hair follicle epithelial cells and hair formation, and it is believed that promoting the proliferation of dermal papilla cells can prevent or improve alopecia. To date, substances known to have the effect of promoting the proliferation of dermal papilla cells include, for example, wild thyme extract (see, for example, Patent Document 23).

[0050] Reactive oxygen species have been attracting attention as a factor that oxidizes biological components, and their adverse effects on the body have become a problem. Reactive oxygen species are generated in the energy metabolism process within living cells, and are produced in the form of superoxide (i.e., superoxide anion (·O2 - ), hydrogen peroxide (H2O2), singlet oxygen ( 1 These reactive oxygen species are essential for the bactericidal mechanism of phagocytes and play an important role in eliminating viruses and cancer cells.

[0051] However, excessive production of reactive oxygen species attacks the biological molecules that make up membranes and tissues in the body, causing various diseases.Superoxide, which is produced in the body and serves as the starting material for other reactive oxygen species, is normally eliminated sequentially by the catalytic action of superoxide dismutase (SOD) contained within cells.

[0052] When superoxide is produced excessively or when SOD activity is reduced, superoxide scavenging is insufficient, resulting in high superoxide concentrations, which is thought to be one of the causes of tissue damage such as rheumatoid arthritis and Behçet's disease, myocardial infarction, stroke, cataracts, age spots, freckles, wrinkles, diabetes, arteriosclerosis, stiff shoulders, sensitivity to cold, and skin aging.

[0053] As described above, various studies have been conducted to date. However, there remains a strong demand for new materials that have at least one of the above-mentioned effects and are highly safe, and therefore can be widely used as ingredients in foods and beverages, cosmetics, research reagents, and the like, and there is currently a demand for their rapid development. [Prior art documents] [Patent documents]

[0054] [Patent Document 1] Japanese Patent Publication No. 2020-094021 [Patent Document 2] Japanese Patent Application Laid-Open No. 2010-235551 [Patent Document 3] Japanese Patent Application Laid-Open No. 2003-321373 [Patent Document 4] Japanese Patent Application Laid-Open No. 2004-107660 [Patent Document 5] Japanese Patent Application Laid-Open No. 2006-063033 [Patent Document 6] Japanese Patent Application Publication No. 10-147515 [Patent Document 7] Japanese Patent Application Laid-Open No. 2004-217618 [Patent Document 8] Japanese Patent Application Laid-Open No. 2007-077169 [Patent Document 9] Japanese Patent Application Laid-Open No. 2005-179243 [Patent Document 10] Japanese Patent Application Laid-Open No. 2000-226308 [Patent Document 11] Japanese Patent Application Laid-Open No. 2007-063160 [Patent Document 12] Japanese Patent Application Laid-Open No. 2006-206571 [Patent Document 13] Japanese Patent Application Laid-Open No. 2009-221110 [Patent Document 14] Japanese Patent Application Laid-Open No. 2010-090035 [Patent Document 15] Japanese Patent Application Laid-Open No. 2007-099698 [Patent Document 16] Japanese Patent Application Laid-Open No. 2006-056854 [Patent Document 17] Japanese Patent Application Laid-Open No. 2009-191039 [Patent Document 18] Japanese Patent Application Laid-Open No. 2007-176830 [Patent Document 19] Japanese Patent Application Laid-Open No. 2007-176835 [Patent Document 20] Japanese Patent Application Laid-Open No. 2009-256272 [Patent Document 21] Japanese Patent Application Laid-Open No. 2010-111615 [Patent Document 22] Japanese Patent Application Laid-Open No. 2003-012532 [Patent Document 23] Japanese Patent Application Laid-Open No. 2006-219407 [Non-patent literature]

[0055] [Non-Patent Document 1] Circulation. 2006;114:2508-2516 [Non-patent document 2] Marinkovich MP et al., "J. Cell. Biol.", 1992, vol. 199, p. 695-703 [Non-patent document 3] Lavker et al., "J. Invest. Dermatol.", 1979, Vol. 73, p. 59-66 [Non-patent document 4] Kaori Miyazaki, "Functional Analysis and Application of Cell Adhesion Molecule Laminin 5", [online], Yokohama City University 3rd Industry-Academia Collaboration Seminar, [Retrieved August 1, 2007], Internet<URL:http: / / www.yokohama‐cu.ac.jp / sangaku / seminar3 / summary_miyazaki.pdf> [Non-Patent Document 5] Takuo Tsuji, Journal of the Japanese Dermatological Association, 1995, Vol. 105, No. 7, pp. 963-975 [Non-patent document 6] "Fragrance Journal", 2006, Vol. 34, No. 10, pp. 19-23 [Non-Patent Document 7] Nat Genet.,2006,Vol.38,No.4,p.441-446 [Non-patent document 8] "Fragrance Journal Special Issue", 2000, Vol. 17, pp. 14-19 [Non-Patent Document 9] Arch. Dermatol. Res., 1996, Vol. 288, p. 442-446 [Non-Patent Document 10] J.Cell Biol.,vol.156,pp.1099-1111(2002) [Non-Patent Document 11] Journal of the Japanese Society of Cosmetic Science, vol.31, pp.296-301(2007) [Non-Patent Document 12] J.Clin.Invest.,vol.99,pp.1272-1280(1997) [Non-Patent Document 13] Kidney Int.,vol.50,pp.1303-1309(1996) [Non-Patent Document 14] J.Biol.Chem.,vol.275,pp.21177-21184(2000) [Non-Patent Document 15] "Pharmacology Atlas", Takehiko Fukuhara, Bunkodo, 1995, p.184 [Non-Patent Document 16] Trends Genet.,1992,Vol.8,Issue 2,p.55-61 Summary of the Invention [Problem to be solved by the invention]

[0056] The present invention aims to solve the above-mentioned conventional problems and achieve the following object: That is, the present invention aims to provide a gout ameliorating agent and a gout ameliorating composition that have an excellent gout ameliorating effect and are highly safe. Another object of the present invention is to provide a liver function improver and a composition for improving liver function that have an excellent liver function improving effect and are highly safe. Another object of the present invention is to provide an immunostimulating agent and a composition for immunostimulation that have excellent immunostimulating activity and are highly safe. Another object of the present invention is to provide an anti-aging agent and an anti-aging composition that have an excellent anti-aging effect and are highly safe. Another object of the present invention is to provide an anti-inflammatory agent and an anti-inflammatory composition that have excellent anti-inflammatory activity and are highly safe. Another object of the present invention is to provide a hair growth agent and a hair growth composition that have an excellent hair growth effect and are highly safe. Another object of the present invention is to provide an antioxidant and an antioxidant composition that have excellent antioxidant activity and are highly safe. [Means for solving the problem]

[0057] As a result of extensive research conducted by the present inventors to solve the above problems, they discovered that Peucedanum japonicum extract has excellent gout-improving effects, liver function-improving effects, immune-stimulating effects, anti-aging effects, anti-inflammatory effects, hair growth effects, and antioxidant effects, is highly safe, and is useful for gout improvement, liver function improvement, immune-stimulating effects, anti-aging effects, anti-inflammation, hair growth, and antioxidant effects, and thus completed the present invention.

[0058] The present invention is based on the above findings of the present inventors, and the means for solving the above problems are as follows: <1> A gout-relieving agent characterized by containing an extract of Peucedanum japonicum. <2> The compound having xanthine oxidase activity inhibitory activity <1> It is a gout improving agent described in the above. <3> The aforementioned <1> from <2> 1. A composition for ameliorating gout, comprising the gout ameliorating agent according to any one of the above. <4> A liver function improver characterized by containing Peucedanum japonicum extract. <5> The compound having at least one of a glutathione production promoting action and an ATP production promoting action. <4> It is a liver function improver described in the above. <6> The aforementioned <4> from <5> 1. A composition for improving liver function, comprising the liver function improver according to any one of the above. <7> This is an immunostimulant characterized by containing an extract of Peucedanum japonicum. <8> The above-mentioned compound has the effect of promoting the production of tumor necrosis factor (TNF-α). <7> The immunostimulant described in the above is an immunostimulant. <9> The aforementioned <7> from <8> The present invention relates to a composition for immunostimulation, characterized by containing the immunostimulant described in any one of the above. <10> This is an anti-aging agent characterized by containing Peucedanum japonicum extract. <11> The above-mentioned compound has at least one of the following effects: type VII collagen production promoting effect, laminin-332 production promoting effect, epidermal keratinocyte proliferation promoting effect, ATP production promoting effect, glutathione production promoting effect, transglutaminase-1 (TGM-1) mRNA expression promoting effect, filaggrin (FLG) mRNA expression promoting effect, aquaporin 3 (AQP3) mRNA expression promoting effect, hyaluronic acid synthase 3 (HAS3) mRNA expression promoting effect, claudin 4 (CLDN4) mRNA expression promoting effect, occludin (OCLN) mRNA expression promoting effect, advanced glycation end products (AGEs) formation inhibiting effect, and advanced glycation end products (AGEs) degradation promoting effect. <10> The anti-aging agent is as described in the above. <12> The aforementioned <10> from <11> The present invention relates to an anti-aging composition comprising the anti-aging agent according to any one of the above items. <13> This is an anti-inflammatory agent characterized by containing an extract of Peucedanum japonicum. <14> The above-mentioned compound having at least one of an inhibitory effect on tumor necrosis factor (TNF-α) production, an inhibitory effect on hexosaminidase release, and an inhibitory effect on prostaglandin (PG) E2 production. <13> It is an anti-inflammatory agent described in <15> The aforementioned <13> from <14> The present invention relates to an anti-inflammatory composition comprising the anti-inflammatory agent according to any one of the above. <16> This hair growth agent is characterized by containing an extract of Peucedanum japonicum. <17> The above-mentioned compound has the effect of promoting proliferation of hair papilla cells. <16> The hair growth agent described in <18> The aforementioned <16> from <17> The hair growth composition is characterized by containing the hair growth agent according to any one of the above items. <19> This is an antioxidant characterized by containing an extract of Peucedanum japonicum. <20> The above-mentioned compound having at least one of a superoxide scavenging activity and a radical scavenging activity. <19> 1. The antioxidant described in <21> The aforementioned <19> from <20> 1. An antioxidant composition comprising the antioxidant described in any one of the above. [Effects of the Invention]

[0059] The gout improving agent and composition for gout improvement of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned object, and can provide a gout improving agent and composition for gout improvement that have excellent gout improving effects and are highly safe. The liver function improver and composition for improving liver function of the present invention can solve the above-mentioned problems of the past and achieve the above-mentioned objectives, and can provide a liver function improver and composition for improving liver function that have excellent liver function improving effects and are highly safe. The immunostimulant and composition for immunostimulation of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned objectives, and can provide an immunostimulant and composition for immunostimulation that have excellent immunostimulating effects and are highly safe. The anti-aging agent and anti-aging composition of the present invention can solve the above-mentioned conventional problems and achieve the above-mentioned object, and can provide an anti-aging agent and anti-aging composition that have excellent anti-aging activity and are highly safe. The anti-inflammatory agent and anti-inflammatory composition of the present invention can solve the above-mentioned conventional problems and achieve the above-mentioned object, and can provide an anti-inflammatory agent and anti-inflammatory composition that have excellent anti-inflammatory enhancing effects and are highly safe. The hair growth agent and hair growth composition of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned object, and can provide a hair growth agent and hair growth composition that has excellent hair growth activity and is highly safe. The antioxidant and antioxidant composition of the present invention can solve the above-mentioned problems of the prior art and achieve the above-mentioned object, and can provide an antioxidant and antioxidant composition that have excellent antioxidant activity and are highly safe. DETAILED DESCRIPTION OF THE INVENTION

[0060] (gout relief agent, liver function improver, immune stimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, and antioxidant) The gout-relieving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, and antioxidant of the present invention contain Peucedanum japonicum extract as an active ingredient, and further contain other ingredients as necessary.

[0061] Although the details of the substances contained in the Peucedanum japonicum extract that exhibit at least one of gout-improving, liver function-improving, immunostimulating, anti-aging, anti-inflammatory, hair-growth, and antioxidant effects are unknown, it was not previously known that the Peucedanum japonicum extract has such excellent effects and is useful as a gout-improving agent, liver function-improving agent, immunostimulating agent, anti-aging agent, anti-inflammatory agent, hair-growth agent, and antioxidant, and this is a new discovery by the present inventors.

[0062] <Peucedanum officinalis extract> The Peucedanum japonicum (scientific name: Peucedanum japonicum Thunb.) is a plant of the genus Umbelliferae in the family Apiaceae, a perennial plant that grows on the coast and is distributed west of the Kanto region, in Shikoku, Kyushu, Okinawa, Korea, mainland China, the Philippines, and other areas, and is easily available in these regions.

[0063] The Peucedanum japonicum extract may be prepared from the part of the plant used as the raw material for extraction, or may be a commercially available product.

[0064] The part of Peucedanum japonicum that can be extracted is not particularly limited and can be appropriately selected depending on the purpose, and examples include above-ground parts, flowers, leaves, stems, rhizomes, roots, seeds, or mixtures thereof. These may be used alone or in combination of two or more. Among these, the roots and above-ground parts are preferred, and the roots are more preferred. The shape, structure and size of the extract material of Peucedanum japonicum are not particularly limited and can be appropriately selected depending on the purpose.

[0065] The method for preparing the extract of Peucedanum japonicum is not particularly limited and can be selected appropriately depending on the purpose. For example, the extract may be dried and then crushed directly or using a crusher. The dried product can be subjected to solvent extraction either directly or after crushing. The drying can be performed in the sun or using a commonly used dryer.

[0066] The Peucedanum japonicum extract can be easily obtained by a method commonly used for plant extraction. The form of the Peucedanum japonicum extract is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include the extract itself, a diluted extract, a concentrated extract, a dried product thereof, a roughly purified product thereof, and a purified product thereof.

[0067] The extraction method is not particularly limited and can be selected appropriately depending on the purpose, and examples include extraction methods using any extraction device at room temperature or under reflux heating. More specifically, an extract can be obtained by placing the extracting part of the Peucedanum japonicum (the raw material) into a treatment tank filled with an extraction solvent, stirring appropriately as necessary, and leaving it to stand for, for example, 30 minutes to 4 hours to elute the soluble components, followed by filtration to remove the extraction residue. The extract can then be dried after distilling off the extraction solvent. Alternatively, the extract may be used as a raw material for extraction after pre-treatment such as degreasing with a non-polar solvent such as hexane. Pre-treatment such as degreasing allows for efficient extraction with a polar solvent.

[0068] The conditions for extracting Peucedanum japonicum (extraction time and extraction temperature), the extraction solvent, and the amount of extraction solvent used are not particularly limited and can be appropriately selected depending on the purpose.

[0069] The extraction solvent is not particularly limited and can be appropriately selected depending on the purpose. Examples thereof include water, a hydrophilic solvent, and a mixed solvent of water and a hydrophilic solvent.

[0070] The water is not particularly limited and can be appropriately selected depending on the purpose. Examples include pure water, tap water, well water, mineral water, hot spring water, spring water, fresh water, and water obtained by various treatments. Examples of treatments for water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, etc. Water that can be used as the extraction solvent also includes purified water, hot water, ion-exchanged water, saline, phosphate buffer, phosphate-buffered saline, etc. The water may be used alone or in combination of two or more types.

[0071] The hydrophilic solvent is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include lower alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin. These may be used alone or in combination of two or more.

[0072] The amount of the hydrophilic solvent used relative to the water in the mixed solvent is not particularly limited and can be appropriately selected depending on the purpose. However, when a lower alcohol is used, it is preferable to add 1 to 90 parts by volume relative to 10 parts by volume of water, when a lower aliphatic ketone is used, it is preferable to add 1 to 40 parts by volume relative to 10 parts by volume of water, and when a polyhydric alcohol is used, it is preferable to add 1 to 90 parts by volume relative to 10 parts by volume of water.

[0073] The temperature of the extraction solvent is not particularly limited and can be appropriately selected depending on the purpose, but it is preferably used at a temperature between room temperature and the boiling point of the solvent.

[0074] The obtained Peucedanum japonicum extract may be subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods to obtain diluted, concentrated, dried, roughly purified, purified, etc. products of the Peucedanum japonicum extract.

[0075] The purification method for the Peucedanum japonicum extract is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, etc. Purification by the above purification methods can increase the concentration of active ingredients and remove unnecessary substances.

[0076] The obtained Peucedanum japonicum extract can be used as it is as a gout alleviating agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant, but the concentrate and dried product are preferred in terms of ease of use. When obtaining the dried product, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity.

[0077] The content of the Peucedanum japonicum extract in the gout ameliorating agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be appropriately adjusted depending on the physiological activity of the extract, etc. The gout ameliorating agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant may consist solely of the Peucedanum japonicum extract.

[0078] <Other ingredients> The other components are not particularly limited and can be appropriately selected depending on the form of use of the gout ameliorating agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant, and examples thereof include excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, coloring agents, fragrances, whitening agents, moisturizers, oily components, UV absorbers, surfactants, thickeners, alcohols, powder components, coloring agents, aqueous components, water, skin nutrients, etc. These may be used alone or in combination of two or more.

[0079] The content of the other ingredients in the gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be selected appropriately depending on the purpose.

[0080] <Application> The use of the gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include pharmaceuticals, quasi-drugs, foods and beverages, and cosmetics. The gout-improving agent, liver function-improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant has at least one of excellent gout-improving effect, liver function-improving effect, immunostimulant effect, anti-aging effect, anti-inflammatory effect, hair growth effect, and antioxidant effect, and is highly safe, and therefore can be suitably used, for example, as an active ingredient of a gout-improving composition, liver function-improving composition, immunostimulant composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition.

[0081] The gout-relieving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective action effects are exerted.

[0082] The method of use of the gout-relieving agent, liver function improver, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include oral, parenteral, and topical use.

[0083] The dosage form of the gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be appropriately selected depending on the purpose. Examples include oral administration agents such as tablets, powders, capsules, granules, extracts, and syrups; parenteral administration agents such as injections, drip infusions, and suppositories; and external preparations such as lotions, emulsions, creams, ointments, beauty serums, lotions, packs, jellies, lip balms, lipsticks, foundations, bath additives, soaps, body soaps, astringents, hair tonics, hair lotions, hair creams, hair liquids, pomades, shampoos, rinses, and conditioners. The method for producing each of the above-mentioned dosage forms of gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited, and any known method can be appropriately selected.

[0084] The method of use, such as the amount used and the period of use, of the gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant is not particularly limited and can be appropriately selected depending on the purpose.

[0085] Furthermore, the gout-improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant of the present invention can also be used as a reagent for research into the mechanism of action of the gout-improving effect, liver function improving effect, immunostimulant effect, anti-aging effect, anti-inflammatory effect, hair growth effect, or antioxidant effect.

[0086] The gout-improving effect of the Peucedanum japonicum extract is exerted, for example, by its xanthine oxidase inhibitory effect. Therefore, the gout-improving agent preferably has a xanthine oxidase inhibitory effect. However, the gout-improving effect of the Peucedanum japonicum extract is not limited to the gout-improving effect exerted based on the xanthine oxidase inhibitory effect. The present invention also relates to a xanthine oxidase activity inhibitor containing the Peucedanum japonicum extract.

[0087] The liver function improving effect of the Peucedanum japonicum extract (e.g., prevention or amelioration of hangovers; prevention, treatment, or amelioration of symptoms caused by the accumulation of harmful substances (rough skin, fatigue, etc.); increased metabolism, etc.) is exerted, for example, by at least one of glutathione production promoting action and ATP production promoting action. Therefore, it is preferable that the liver function improver has at least one of glutathione production promoting action and ATP production promoting action. Note that the liver function improving effect of the Peucedanum japonicum extract is not limited to the liver function improving action exerted based on at least one of glutathione production promoting action and ATP production promoting action. The present invention also relates to a glutathione production promoter or an ATP production promoter containing the Peucedanum japonicum extract.

[0088] The immunopotentiating effect of the Peucedanum japonicum extract is exerted, for example, by promoting the production of tumor necrosis factor (TNF-α). Therefore, the immunopotentiator preferably has the effect of promoting the production of tumor necrosis factor (TNF-α). However, the immunopotentiating effect of the Peucedanum japonicum extract is not limited to the immunopotentiating effect exerted based on the effect of promoting the production of tumor necrosis factor (TNF-α). The present invention also relates to a tumor necrosis factor (TNF-α) production promoter containing the Peucedanum japonicum extract.

[0089] The anti-aging effect of the Peucedanum japonicum extract is exerted by at least one of the following actions: promoting type VII collagen production, promoting laminin-332 production, promoting epidermal keratinocyte proliferation, promoting ATP production, promoting glutathione production, promoting transglutaminase-1 (TGM-1) mRNA expression, promoting filaggrin (FLG) mRNA expression, promoting aquaporin 3 (AQP3) mRNA expression, promoting hyaluronic acid synthase 3 (HAS3) mRNA expression, promoting claudin 4 (CLDN4) mRNA expression, promoting occludin (OCLN) mRNA expression, inhibiting the formation of advanced glycation end products (AGEs), and promoting the degradation of advanced glycation end products (AGEs). Therefore, it is preferable that the anti-aging agent has at least one of the following effects: type VII collagen production promotion, laminin-332 production promotion, epidermal keratinocyte proliferation promotion, ATP production promotion, glutathione production promotion, transglutaminase-1 (TGM-1) mRNA expression promotion, filaggrin (FLG) mRNA expression promotion, aquaporin 3 (AQP3) mRNA expression promotion, hyaluronic acid synthase 3 (HAS3) mRNA expression promotion, claudin 4 (CLDN4) mRNA expression promotion, occludin (OCLN) mRNA expression promotion, advanced glycation end products (AGEs) formation inhibition, and advanced glycation end products (AGEs) degradation promotion. The anti-aging effects of the Peucedanum japonicum extract are not limited to those exerted based on at least one of the following: type VII collagen production promotion, laminin-332 production promotion, epidermal keratinocyte proliferation promotion, ATP production promotion, glutathione production promotion, transglutaminase-1 (TGM-1) mRNA expression promotion, filaggrin (FLG) mRNA expression promotion, aquaporin 3 (AQP3) mRNA expression promotion, hyaluronic acid synthase 3 (HAS3) mRNA expression promotion, claudin 4 (CLDN4) mRNA expression promotion, occludin (OCLN) mRNA expression promotion, advanced glycation end products (AGEs) formation inhibition, and advanced glycation end products (AGEs) degradation promotion. The present invention also relates to a type VII collagen production promoter, a laminin-332 production promoter, an epidermal keratinocyte proliferation promoter, an ATP production promoter, a glutathione production promoter, a transglutaminase-1 (TGM-1) mRNA expression promoter, a filaggrin (FLG) mRNA expression promoter, an aquaporin 3 (AQP3) mRNA expression promoter, a hyaluronan synthase 3 (HAS3) mRNA expression promoter, a claudin 4 (CLDN4) mRNA expression promoter, an occludin (OCLN) mRNA expression promoter, an advanced glycation end products (AGEs) formation inhibitor, or an advanced glycation end products (AGEs) degradation promoter, each containing the above-mentioned Peucedanum extract.

[0090] The anti-inflammatory effect of the Peucedanum japonicum extract is exerted, for example, by at least one of the following: suppression of tumor necrosis factor (TNF-α) production, suppression of hexosaminidase release, and suppression of prostaglandin (PG) E2 production. Therefore, the anti-inflammatory agent preferably has at least one of the following: suppression of tumor necrosis factor (TNF-α) production, suppression of hexosaminidase release, and suppression of prostaglandin (PG) E2 production. Note that the anti-inflammatory effect of the Peucedanum japonicum extract is not limited to the anti-inflammatory effect exerted based on at least one of the following: suppression of tumor necrosis factor (TNF-α) production, suppression of hexosaminidase release, and suppression of prostaglandin (PG) E2 production. The present invention also relates to a tumor necrosis factor (TNF-α) production inhibitor, a hexosaminidase release inhibitor, or a prostaglandin (PG) E2 production inhibitor, which contains the Peucedanum japonicum extract.

[0091] The hair growth effect of the Peucedanum japonicum extract is exerted, for example, by promoting the proliferation of hair papilla cells. Therefore, the hair growth agent preferably has the action of promoting the proliferation of hair papilla cells. However, the hair growth effect of the Peucedanum japonicum extract is not limited to the hair growth effect exerted based on the action of promoting the proliferation of hair papilla cells. The present invention also relates to a hair papilla cell proliferation promoter containing the Peucedanum japonicum extract.

[0092] The antioxidant effect of the Peucedanum japonicum extract is exerted, for example, by at least one of a superoxide scavenging effect and a radical scavenging effect. Therefore, it is preferable that the antioxidant has at least one of a superoxide scavenging effect and a radical scavenging effect. However, the antioxidant effect of the Peucedanum japonicum extract is not limited to the antioxidant effect exerted based on at least one of a superoxide scavenging effect and a radical scavenging effect. The present invention also relates to a superoxide scavenger or radical scavenger containing the Peucedanum japonicum extract.

[0093] (Compositions for improving gout, compositions for improving liver function, compositions for stimulating immunity, compositions for anti-aging, compositions for anti-inflammation, compositions for hair growth, and compositions for antioxidants) The composition for ameliorating gout of the present invention contains the gout ameliorating agent of the present invention, and further contains other ingredients as required. The composition for improving liver function of the present invention contains the liver function improver of the present invention, and further contains other ingredients as necessary. The immunostimulating composition of the present invention contains the immunostimulating agent of the present invention and further contains other components as necessary. The anti-aging composition of the present invention contains the anti-aging agent of the present invention and further contains other components as required. The anti-inflammatory composition of the present invention contains the anti-inflammatory agent of the present invention and further contains other ingredients as required. The hair growth composition of the present invention contains the hair growth agent of the present invention and may further contain other ingredients as required. The antioxidant composition of the present invention contains the antioxidant of the present invention and further contains other components as required.

[0094] <Gout relief agent, liver function improver, immune stimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, and antioxidant> The gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, and antioxidant are the gout improving agent, liver function improving agent, immunostimulant, anti-aging agent, anti-inflammatory agent, hair growth agent, and antioxidant of the present invention described above.

[0095] The content of the gout improving agent, liver function improving agent, immunostimulating agent, anti-aging agent, anti-inflammatory agent, hair growth agent, or antioxidant in the gout improving composition, liver function improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition is not particularly limited and can be appropriately adjusted depending on the form of the gout improving composition, liver function improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition, the physiological activity of the Peucedanum japonicum extract, etc., but is preferably 0.0001% by mass to 20% by mass, and more preferably 0.0001% by mass to 10% by mass, in terms of the Peucedanum japonicum extract. The gout-improving composition, liver function-improving composition, immune-stimulating composition, anti-aging composition, anti-inflammatory composition, hair-growth composition, or antioxidant composition may consist solely of the gout-improving agent, liver function-improving agent, immune-stimulating agent, anti-aging agent, anti-inflammatory agent, hair-growth agent, or antioxidant.

[0096] <Other ingredients> The other components in the gout-ameliorating composition, liver function-improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition are not particularly limited and can be appropriately selected depending on the form of use of the gout-ameliorating composition, liver function-improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition, and examples thereof include the same as the other components described in the above-mentioned items for gout-ameliorating agents, liver function-improving agents, immunostimulating agents, anti-aging agents, anti-inflammatory agents, hair growth agents, or antioxidants. These may be used alone or in combination of two or more.

[0097] The content of the other ingredients in the composition for improving gout, composition for improving liver function, composition for immunostimulation, composition for anti-aging, composition for anti-inflammatory, composition for hair growth, or composition for antioxidant purposes is not particularly limited and can be selected appropriately depending on the purpose.

[0098] <Aspect> The form of the composition for improving gout, composition for improving liver function, composition for immunostimulation, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include pharmaceuticals, quasi-drugs, foods and beverages, cosmetics, etc. The composition for improving gout, improving liver function, stimulating immunity, anti-aging, anti-inflammatory, hair growth, or antioxidant of the present invention can be used on a daily basis, and due to the action of the active ingredient, Peucedanum japonicum extract, it can extremely effectively exert various physiologically active effects including gout improvement, improving liver function, stimulating immunity, anti-aging, anti-inflammatory, hair growth, or antioxidant effects.

[0099] The composition for improving gout, composition for improving liver function, composition for immune stimulation, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective functional effects are exerted.

[0100] The method of use of the composition for improving gout, composition for improving liver function, composition for immunostimulation, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition of the present invention is not particularly limited and can be appropriately selected depending on the purpose, and examples include oral, parenteral, and topical use.

[0101] Examples of the oral composition include the above-mentioned orally administered agents and foods and beverages. Here, foods and beverages refer to those that are unlikely to be harmful to human health and are taken orally or by administration through the digestive tract in normal social life, and are not limited to administrative classifications such as foods, medicines, and quasi-drugs. Therefore, the foods and beverages refer to a wide range of foods and beverages that are taken orally, including general foods, health foods (functional foods and beverages), health functional foods (foods for specified health uses, foods with nutrient functions, and foods with functional claims), quasi-drugs, and pharmaceuticals.

[0102] The type of oral composition is not particularly limited and can be appropriately selected depending on the purpose. Examples of the oral composition include beverages such as tea drinks, soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, alcoholic drinks, coffee drinks, and coffee-containing soft drinks (including concentrated liquids and powders for adjusting these beverages); frozen desserts such as ice cream, ice sorbet, and shaved ice; noodles such as soba noodles, udon noodles, vermicelli, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; confectioneries such as candy, candy, gum, chocolate, candy tablets, snacks, biscuits, jelly, jam, cream, baked goods, and bread; and seafood such as crab, salmon, clams, tuna, sardines, shrimp, bonito, mackerel, whale, oysters, saury, squid, ark shells, scallops, abalone, sea urchin, salmon roe, and tokobushi sea bream. ; Processed seafood and livestock foods such as kamaboko, ham, and sausage; Dairy products such as processed milk and fermented milk; Oils and fats and oil-processed foods such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressings; Condiments such as sauces and dressings; Retort pouch foods such as curry, stew, oyakodon, porridge, rice porridge, Chinese rice bowl, katsudon, tendon, unadon, hayashi rice, oden, mabo dolph, beef bowl, meat sauce, egg soup, omelet rice, gyoza, shumai, hamburger steak, and meatballs; Side dishes such as salads and pickles; Health, beauty, and nutritional supplements in various forms; Pharmaceuticals and quasi-drugs such as tablets, granules, capsules, drinks, lozenges, and mouthwash; Oral fresheners used in the mouth such as mouth fresheners and anti-halitosis agents, and toothpaste.

[0103] Examples of the non-oral compositions include the above-mentioned parenteral preparations and topical preparations, such as ointments, creams, emulsions, lotions, packs, and foundations that can be used as skin cosmetics, and hair tonics, hair creams, hair liquids, shampoos, pomades, and rinses that can be used as hair cosmetics.

[0104] The method for producing the gout-improving composition, liver function-improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition is not particularly limited, and can be appropriately selected depending on the form in which the gout-improving composition, liver function-improving composition, immunostimulating composition, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition is to be used.

[0105] The amount, duration of use, etc. of the composition for improving gout, composition for improving liver function, composition for immunostimulation, anti-aging composition, anti-inflammatory composition, hair growth composition, or antioxidant composition are not particularly limited and can be selected appropriately depending on the purpose.

[0106] As described above, the gout-improving agent and gout-improving composition of the present invention have an excellent gout-improving effect, the liver function-improving agent and liver function-improving composition have an excellent liver function-improving effect, the immunostimulant and immunostimulating composition have an excellent immunostimulating effect, the anti-aging agent and anti-aging composition have an excellent anti-aging effect, the anti-inflammatory agent and anti-inflammatory composition have an excellent anti-inflammatory effect, the hair growth agent and hair growth composition have an excellent hair growth effect, and the antioxidant and antioxidant composition have an excellent antioxidant effect. Therefore, the present invention also relates to a method for improving gout, which comprises administering at least one of the gout improving agent and gout improving composition to an individual; a method for improving liver function, which comprises administering at least one of the liver function improving agent and liver function improving composition to an individual; an immunostimulating method, which comprises administering at least one of the immunostimulating agent and immunostimulating composition to an individual; an anti-aging method, which comprises administering at least one of the anti-aging agent and anti-aging composition to an individual; an anti-inflammatory method, which comprises administering at least one of the anti-inflammatory agent and anti-inflammatory composition to an individual; a hair growth method, which comprises administering at least one of the hair growth agent and hair growth composition to an individual; and an antioxidant method, which comprises administering at least one of the antioxidant and antioxidant composition to an individual. [Example]

[0107] Hereinafter, production examples, test examples and formulation examples of the present invention will be explained, but the present invention is not limited to these production examples, test examples and formulation examples.

[0108] (Production Example 1) An extract of the roots of Peucedanum japonicum with 30% by volume ethanol was prepared as follows. 200 mL of 30% by volume ethanol was added to 10 g of Peucedanum japonicum root, and extraction was carried out at 80°C for 1 hour using a reflux condenser, and then filtered through filter paper to obtain the extract. The obtained extract was concentrated under reduced pressure and dried to obtain 2.28 g of a 30% by volume ethanol extract (powder) of the roots of Peucedanum japonicum.

[0109] (Production Example 2) 2.23 g of a 50% by volume ethanol extract (powder) of the roots of Peucedanum japonicum was obtained in the same manner as in Production Example 1, except that 30% by volume ethanol was replaced with 50% by volume ethanol.

[0110] (Production Example 3) 1.90 g of an 80% by volume ethanol extract (powder) of the roots of Peucedanum japonicum was obtained in the same manner as in Production Example 1, except that 30% by volume ethanol was replaced with 80% by volume ethanol.

[0111] (Production Example 4) In Production Example 1, 10 g of the root parts of Peucedanum japonicum were replaced with 30 g of the above-ground parts of Peucedanum japonicum, and 200 mL of 30% ethanol by 600 mL of 30% ethanol by volume was replaced with 600 mL of 30% ethanol by volume. The same procedure as in Production Example 1 was repeated to obtain 10.48 g of 30% ethanol extract (powder) of the above-ground parts of Peucedanum japonicum.

[0112] (Production Example 5) 10.28 g of a 50% by volume ethanol extract (powder) of the aerial parts of Peucedanum japonicum was obtained in the same manner as in Production Example 4, except that 30% by volume ethanol was replaced with 50% by volume ethanol.

[0113] (Production Example 6) 9.27 g of an 80% by volume ethanol extract (powder) of the aerial parts of Peucedanum japonicum was obtained in the same manner as in Production Example 4, except that 30% by volume ethanol was replaced with 80% by volume ethanol.

[0114] (Test Example 1: Xanthine oxidase activity inhibitory effect test) The extract of Peucedanum japonicum was used as a test sample and its inhibitory effect on xanthine oxidase activity was tested by the following test method.

[0115] 75 μL of 0.1 mol / L phosphate buffer (pH 7.4), 40 μL of distilled water, 5 μL of methanol, and 5 μL of test sample solution (final concentrations see Tables 1-1 and 1-2 below) were added to a 96-well quartz microplate, to which 25 μL of xanthine oxidase solution (0.12 U / mL) was added and heated at 37°C for 15 minutes. After heating, 150 μL of 0.1 mol / L xanthine solution was added, stirred quickly, and incubated at 37°C for 15 minutes. After the reaction, absorbance at a wavelength of 295 nm was measured. As a control, the same procedure was performed using only solvent (control solution) without the test sample. From the obtained results, the xanthine oxidase activity inhibition rate was calculated using the following formula: The results are shown in Tables 1-1 and 1-2. Xanthine oxidase activity inhibition rate (%) = 100 - {(AB) / (CD) × 100} In the above formula, A to D respectively represent the following. A: Absorbance at a wavelength of 295 nm when test sample and enzyme solution are added B: Absorbance at a wavelength of 295 nm with test sample added and without enzyme solution added C: Absorbance at 295 nm when no test sample was added (control) and when enzyme solution was added D: Absorbance at a wavelength of 295 nm without the addition of test sample (control) or enzyme solution

[0116] [Table 1-1]

[0117] [Table 1-2]

[0118] The results in Tables 1-1 and 1-2 confirm that the Peucedanum extract has excellent inhibitory effect on xanthine oxidase activity.

[0119] (Test Example 2: Glutathione production promoting effect test (human normal hepatocytes)) The extract of Peucedanum japonicum was used as a test sample to test its glutathione production promoting effect in normal human hepatocytes according to the following test method.

[0120] Normal human hepatocytes were cultured in Dulbecco's MEM medium containing 10% FBS, and then harvested by trypsinization. The harvested cells were collected at a concentration of 1.0 × 10 5 After diluting with Dulbecco's MEM medium containing 10% FBS to a concentration of 100 cells / mL, 200 μL was seeded per well of a 48-well plate and cultured overnight. After overnight incubation, 200 μL of test sample dissolved in Dulbecco's MEM medium containing 1% FBS (see Tables 2-1 and 2-2 below for sample concentrations) was added to each well and incubated for an additional 24 hours. As a control, cells were incubated in Dulbecco's MEM medium containing 1% FBS without the test sample. After the incubation, the medium was removed from each well, and the wells were washed with 400 μL of PBS(-) buffer, after which the cells were lysed using 150 μL of M-PER (PIERCE). Total glutathione was quantified using 100 μL of this solution. Specifically, 100 μL of lysed cell extract, 50 μL of 0.1 mol / L phosphate buffer, 25 μL of 2 mmol / L NADPH, and 25 μL of 3.2 units / mL glutathione reductase were added to a 96-well plate and incubated at 37°C for 10 minutes. After incubation, 25 μL of 10 mmol / L 5,5'-dithiobis(2-nitrobenzoic acid) was added. The absorbance at 412 nm was measured for 5 minutes, and the ΔOD / min was calculated. Total glutathione concentrations were calculated based on a calibration curve prepared using oxidized glutathione. The obtained values ​​were corrected to the amount of glutathione per total protein amount, and then the glutathione production promotion rate (%) was calculated using the following formula: The results are shown in Tables 2-1 and 2-2. Glutathione production promotion rate (%) = B / A x 100 In the above formula, A and B respectively represent the following. A: Amount of glutathione per total protein in cells without the addition of the test sample (control) B: Amount of glutathione per total protein in cells when the test sample was added

[0121] [Table 2-1]

[0122] [Table 2-2]

[0123] The results in Tables 2-1 and 2-2 confirm that the Peucedanum extract has an excellent effect of promoting glutathione production in hepatocytes.

[0124] (Test Example 3: ATP production promoting effect test using normal human hepatocytes) The Peucedanum japonicum extract was used as a test sample and its ATP production promoting effect was tested by the following test method.

[0125] Normal human hepatocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS, and then harvested by trypsinization. The harvested cells were collected at a concentration of 2.0 × 10 5 After diluting with DMEM containing 10% FBS to a concentration of 100 cells / mL, 100 μL was seeded per well of a 96-well plate and cultured overnight. After overnight incubation, the medium was removed, and 100 μL of the test sample dissolved in 10% FBS-containing DMEM (see Tables 3-1 and 3-2 below for sample concentrations) was added to each well and incubated for 2 hours. As a control, cells were incubated in the same manner using 10% FBS-containing DMEM without the test sample. The ATP production promoting effect was measured using the firefly luciferase luminescence assay to measure the amount of intracellular ATP. Specifically, after incubation, 100 μL of "Cellular" ATP Measurement Reagent (manufactured by Toyo B-Net Co., Ltd.) was added to each well, and a luciferase-mediated chemiluminescence reaction was performed. After the reaction, the amount of chemiluminescence, which was proportional to the amount of intracellular ATP, was measured using a chemiluminescence analyzer (manufactured by ThermoFisher Scientific, product name: Varioskan LUX). From the obtained results, the ATP production promotion rate (%) was calculated using the following formula: The results are shown in Tables 3-1 and 3-2. ATP production promotion rate (%)=A / B×100 In the above formula, A and B respectively represent the following. A: Amount of chemiluminescence in cells when test sample is added B: Amount of chemiluminescence in cells without adding test sample

[0126] [Table 3-1]

[0127] [Table 3-2]

[0128] The results in Tables 3-1 and 3-2 confirm that the Peucedanum extract has an excellent effect of promoting ATP production in hepatocytes.

[0129] (Test Example 4: Test of tumor necrosis factor (TNF-α) production promoting effect) The Peucedanum japonicum extract was used as a test sample and its TNF-α production promoting effect was tested by the following test method.

[0130] Mouse macrophage cells (RAW264.7) were cultured in Dulbecco's MEM containing 10% FBS, and then the cells were collected using a cell scraper. The collected cells were collected at a concentration of 1.0 × 10 6After diluting with Dulbecco's MEM containing 10% FBS to a concentration of 100 cells / mL, 100 μL was seeded per well of a 96-well plate and cultured for 4 hours. After incubation, 100 μL of the test sample (see Tables 4-1 and 4-2 below for concentrations) dissolved in 10% FBS-containing Dulbecco's MEM containing 0.5% DMSO was added to each well and incubated for 24 hours. The control was treated in the same manner except that the test sample was not added. After the culture was completed, the amount of TNF-α in the culture supernatant of each well was measured by sandwich ELISA. From the obtained results, the TNF-α production promotion rate was calculated using the following formula: The results are shown in Tables 4-1 and 4-2. TNF-α production promotion rate (%)=A / B×100 In the above formula, A and B respectively represent the following. A: Amount of TNF-α when test sample was added B: Amount of TNF-α without addition of test sample

[0131] [Table 4-1]

[0132] [Table 4-2]

[0133] The results in Tables 4-1 and 4-2 confirm that the Peucedanum extract has an excellent effect of promoting TNF-α production.

[0134] (Test Example 5: Test of type VII collagen production promoting effect) The Peucedanum japonicum extract was used as a test sample and tested for its type VII collagen production promoting effect by the following test method.

[0135] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were collected at a concentration of 1 × 105 After diluting with KGM without BPE (KGM-BPE) to a concentration of 100 cells / mL, 500 μL was seeded per well of a 24-well plate and cultured for 1 day. After incubation, the medium was removed, and 500 μL of the test sample dissolved in KGM-BPE (see Table 5 below for sample concentrations) or KGM-BPE without the test sample (control) was added to each well and incubated for 48 hours. After incubation, the amount of type VII collagen in the medium in each well was measured by ELISA. From the obtained results, the promotion rate of type VII collagen production was calculated using the following formula. The results are shown in Table 5. Type VII collagen production promotion rate (%) = A / B x 100 In the above formula, A and B respectively represent the following. A: Absorbance at a wavelength of 405 nm when the test sample is added B: Absorbance at 405 nm when no test sample is added (control)

[0136] [Table 5]

[0137] The results in Table 5 confirm that the Peucedanum japonicum extract has an excellent effect of promoting type VII collagen production.

[0138] (Test Example 6: Test of laminin-332 (laminin-5) production promoting effect) The Peucedanum japonicum extract was used as a test sample and tested for its laminin-332 (laminin-5) production promoting effect by the following test method.

[0139] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were collected at a concentration of 1.0 × 10 5 After dilution with BPE-free KGM (KGM-BPE) to a concentration of 100 cells / mL, 500 μL was seeded per well of a 24-well plate and cultured for 1 day. After incubation, the medium was removed, and 500 μL of test sample dissolved in KGM-BPE (concentrations shown in Tables 6-1 and 6-2 below) or KGM-BPE without test sample (control) was added to each well and incubated for 48 hours. After incubation, the amount of laminin-332 in the medium in each well was measured by ELISA. From the obtained results, the laminin-332 production promotion rate was calculated using the following formula: The results are shown in Tables 6-1 and 6-2. Laminin-332 production promotion rate (%) = A / B x 100 In the above formula, A and B respectively represent the following. A: Absorbance at a wavelength of 405 nm when the test sample is added B: Absorbance at 405 nm when no test sample is added (control)

[0140] [Table 6-1]

[0141] [Table 6-2]

[0142] The results in Tables 6-1 and 6-2 confirm that the Peucedanum extract has an excellent effect of promoting the production of laminin-332 (laminin-5).

[0143] (Test Example 7: Epidermal keratinocyte proliferation promoting activity test) The Peucedanum japonicum extract was used as a test sample and tested for its epidermal keratinocyte proliferation promoting effect by the following test method.

[0144] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were collected at a concentration of 3.0 × 10 4 After diluting with KGM to a concentration of 100 cells / mL, the cells were seeded in a collagen-coated 96-well plate at 100 μL per well and cultured overnight. After incubation, 100 μL of the test sample dissolved in KGM (see Tables 7-1 and 7-2 below for concentrations) was added to each well and incubated for 3 days. As a control, incubation was performed in the same manner using KGM without the test sample. The activity of promoting epidermal keratinocyte proliferation was measured using the MTT assay. After 3 days of culture, the medium was removed, and 100 μL of MTT dissolved in PBS(-) buffer at a final concentration of 0.4 mg / mL was added to each well. After 2 hours of culture, the blue formazan produced within the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at 570 nm was measured. At the same time, the absorbance at 650 nm was measured as turbidity, and the difference between the two was used to determine the amount of blue formazan produced. From the obtained results, the epidermal keratinocyte proliferation promotion rate was calculated using the following formula: The results are shown in Tables 7-1 and 7-2. Epidermal keratinocyte proliferation promotion rate (%) = A / B x 100 In the above formula, A and B respectively represent the following. A: Amount of blue formazan produced in cells when test sample was added B: Amount of blue formazan produced in cells without the addition of test sample

[0145] [Table 7-1]

[0146] [Table 7-2]

[0147] The results in Tables 7-1 and 7-2 confirm that the Peucedanum extract has an excellent effect of promoting the proliferation of epidermal keratinocytes.

[0148] (Test Example 8: ATP production promoting effect test) The Peucedanum japonicum extract was used as a test sample to test its ATP production promoting effect in epidermal keratinocytes according to the following test method.

[0149] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were collected at a concentration of 2.0 × 10 5 After diluting with KGM to a concentration of 100 cells / mL, the cells were seeded in a collagen-coated 96-well plate at 100 μL per well and cultured overnight. After overnight incubation, the medium was removed, and 100 μL of the test sample dissolved in KGM (see Table 8 below for sample concentration) was added to each well and incubated for 2 hours. As a control, KGM without the test sample was used and incubated in the same manner. The ATP production promoting effect was measured using the firefly luciferase luminescence assay to measure the amount of intracellular ATP. Specifically, after incubation, 100 μL of "Cellular" ATP Measurement Reagent (manufactured by Toyo B-Net Co., Ltd.) was added to each well, and a luciferase-mediated chemiluminescence reaction was performed. After the reaction, the amount of chemiluminescence, which was proportional to the amount of intracellular ATP, was measured using a chemiluminescence analyzer (manufactured by ThermoFisher Scientific, product name: Varioskan LUX). From the obtained results, the ATP production promotion rate (%) was calculated using the following formula: The results are shown in Table 8. ATP production promotion rate (%)=A / B×100 In the above formula, A and B respectively represent the following. A: Amount of chemiluminescence in cells when test sample is added B: Amount of chemiluminescence in cells without adding test sample

[0150] [Table 8]

[0151] The results in Table 8 confirm that the Peucedanum extract has an excellent effect of promoting ATP production in epidermal keratinocytes.

[0152] (Test Example 9: Glutathione production promoting effect test (fibroblasts)) The extract of Peucedanum japonicum was used as a test sample and its effect of promoting glutathione production in fibroblasts was tested by the following test method.

[0153] Normal human dermal fibroblasts (NB1RGB) were cultured in α-MEM containing 10% FBS, and then harvested by trypsinization. The harvested cells were collected at a concentration of 2.0 × 10 5 After diluting with α-MEM containing 10% FBS to a concentration of 100 cells / mL, 200 μL was seeded per well of a 48-well plate and cultured for 48 hours. After incubation, 200 μL of test sample dissolved in 1% FBS-containing DMEM (see Tables 9-1 and 9-2 below for sample concentrations) was added to each well and incubated for an additional 24 hours. As a control, incubation was performed in the same manner using 1% FBS-containing DMEM without the test sample. After the incubation, the medium was removed from each well, and the wells were washed with 300 μL of PBS(-) buffer, after which the cells were lysed using 150 μL of M-PER (PIERCE). Total glutathione was quantified using 100 μL of this solution. Specifically, 100 μL of lysed cell extract, 50 μL of 0.1 mol / L phosphate buffer, 25 μL of 2 mmol / L NADPH, and 25 μL of 3.2 units / mL glutathione reductase were added to a 96-well plate and incubated at 37°C for 10 minutes. After incubation, 25 μL of 10 mmol / L 5,5'-dithiobis(2-nitrobenzoic acid) was added. The absorbance at 412 nm was measured for 5 minutes, and the ΔOD / min was calculated. Total glutathione concentrations were calculated based on a calibration curve prepared using oxidized glutathione. The obtained values ​​were corrected to the amount of glutathione per total protein amount, and the glutathione production promotion rate (%) was calculated using the following formula: The results are shown in Tables 9-1 and 9-2. Glutathione production promotion rate (%) = B / A x 100 In the above formula, A and B respectively represent the following. A: Amount of glutathione per total protein in cells without the addition of the test sample (control) B: Amount of glutathione per total protein in cells when the test sample was added

[0154] [Table 9-1]

[0155] [Table 9-2]

[0156] The results in Tables 9-1 and 9-2 confirm that the Peucedanum extract has an excellent effect of promoting glutathione production in skin fibroblasts.

[0157] (Test Example 10: Transglutaminase-1 (TGM-1) mRNA expression promoting effect test) The Peucedanum japonicum extract was used as a test sample and tested for its promoting effect on TGM-1 mRNA expression by the following test method.

[0158] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 10-1 and 10-2 below for concentrations) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of TGM-1 mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript. TM Two-step real-time RT-PCR was performed using RT Master Mix (Perfect Real Time) and TB Green® Fast qPCR Mix (Takara). The expression level of TGM-1 mRNA was calculated after correcting it with the expression level of GAPDH mRNA. From the obtained results, the TGM-1 mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 10-1 and 10-2. TGM-1 mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0159] [Table 10-1]

[0160] [Table 10-2]

[0161] The results in Tables 10-1 and 10-2 confirmed that the Peucedanum extract had an excellent effect of promoting TGM-1 mRNA expression.

[0162] (Test Example 11: Filaggrin (FLG) mRNA expression promoting effect test) The Peucedanum japonicum extract was used as a test sample and tested for its FLG mRNA expression promoting effect by the following test method.

[0163] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 11-1 and 11-2 below for concentrations) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of FLG and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript. TM Two-step real-time RT-PCR was performed using RT Master Mix (Perfect Real Time) and TB Green® Fast qPCR Mix (Takara). The expression level of FLG mRNA was calculated after correcting it with the expression level of GAPDH mRNA. From the obtained results, the FLG mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 11-1 and 11-2. FLG mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0164] [Table 11-1]

[0165] [Table 11-2]

[0166] The results in Tables 11-1 and 11-2 confirmed that the Peucedanum japonicum extract had an excellent FLG mRNA expression promoting effect.

[0167] (Test Example 12: Aquaporin 3 (AQP3) mRNA expression promoting effect test) The Peucedanum japonicum extract was used as a test sample and tested for its AQP3 mRNA expression promoting effect by the following test method.

[0168] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 12-1 and 12-2 below for concentrations) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of AQP3 mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript. TMA two-step real-time RT-PCR reaction was performed using RT Master Mix (Perfect Real Time) and TB Green® Fast qPCR Mix (Takara). The expression level of AQP3 mRNA was calculated after normalizing it with the expression level of GAPDH mRNA. From the obtained results, the AQP3 mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 12-1 and 12-2. AQP3 mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0169] [Table 12-1]

[0170] [Table 12-2]

[0171] The results in Tables 12-1 and 12-2 confirmed that the Peucedanum japonicum extract had an excellent effect of promoting AQP3 mRNA expression.

[0172] (Test Example 13: Hyaluronic acid synthase 3 (HAS3) mRNA expression promoting effect test) The Peucedanum japonicum extract was used as a test sample and tested for its promoting effect on HAS3 mRNA expression by the following test method.

[0173] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 13-1 and 13-2 below for concentrations) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of HAS3 mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript PCR. TM Two-step real-time RT-PCR was performed using RT Master Mix (Perfect Real Time) and TB Green® Fast qPCR Mix (Takara). The expression level of HAS3 mRNA was calculated after normalizing it with the expression level of GAPDH mRNA. From the obtained results, the HAS3 mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 13-1 and 13-2. HAS3 mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0174] [Table 13-1]

[0175] [Table 13-2]

[0176] The results in Tables 13-1 and 13-2 confirmed that the Peucedanum japonicum extract had an excellent effect of promoting HAS3 mRNA expression.

[0177] (Test Example 14: Claudin 4 (CLDN4) mRNA expression promoting effect test) The Peucedanum japonicum extract was used as a test sample and tested for its promoting effect on CLDN4 mRNA expression by the following test method.

[0178] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL / well and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 14-1 and 14-2 below) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of CLDN4 mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript PCR. TM Two-step real-time RT-PCR was performed using RT Master Mix (Perfect Real Time) and TB Green (registered trademark) Fast qPCR Mix (Takara). The expression level of CLDN4 mRNA was calculated after normalizing it with the expression level of GAPDH mRNA. From the obtained results, the CLDN4 mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 14-1 and 14-2. CLDN4 mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0179] [Table 14-1]

[0180] [Table 14-2]

[0181] The results in Tables 14-1 and 14-2 confirmed that the Peucedanum japonicum extract had an excellent effect of promoting CLDN4 mRNA expression.

[0182] (Test Example 15: Test of occludin (OCLN) mRNA expression promoting effect) The extract of Peucedanum japonicum was used as a test sample and its effect of promoting OCLN mRNA expression was tested by the following test method.

[0183] Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were plated in 6-well plates at 3.0 × 10 5 The cells were seeded at 2 mL each and cultured overnight at 37°C under 5% CO2. After overnight incubation, the medium was replaced with growth factor-free KBM. After 24 hours, the culture medium was discarded, and 2 mL of test sample dissolved in KBM at the required concentration (see Tables 15-1 and 15-2 below) was added to each well and incubated at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was used and incubated in the same manner. After the culture, the medium was removed, total RNA was extracted using ISOGEN II (Nippon Gene), the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and total RNA was adjusted to 200 ng / μL. Using this total RNA as a template, the expression levels of OCLN mRNA and GAPDH mRNA (an internal standard) were measured. Detection was performed using a real-time PCR device, Thermal Cycler Dice® Real Times System III (manufactured by Takara) with PrimeScript. TM Two-step real-time RT-PCR was performed using RT Master Mix (Perfect Real Time) and TB Green® Fast qPCR Mix (Takara). The expression level of OCLN mRNA was calculated after correcting it with the expression level of GAPDH mRNA. From the obtained results, the OCLN mRNA expression promotion rate was calculated using the following formula: The results are shown in Tables 15-1 and 15-2. OCLN mRNA expression promotion rate (%) = A / B × 100 In the above formula, A and B respectively represent the following. A: Correction value when test sample is added B: Corrected value when no test sample is added

[0184] [Table 15-1]

[0185] [Table 15-2]

[0186] The results in Tables 15-1 and 15-2 confirmed that the Peucedanum japonicum extract had an excellent effect of promoting OCLN mRNA expression.

[0187] (Test Example 16: Test of inhibitory effect on the formation of advanced glycation end products (AGEs)) The Peucedanum japonicum extract was used as a test sample and its inhibitory effect on AGE formation was tested by the following test method.

[0188] 100 μL of a mixture of 0.2 mol / L D(-)-ribose prepared in PBS(-) and various concentrations of test samples (see Tables 16-1 and 16-2 below for concentrations) was added to a 96-well type I collagen-coated plate and left to stand at 37°C for 20 days to allow AGEs to form. At this time, a plate to which only PBS(-) was added was used as a negative control, and a plate to which only D(-)-ribose was added was used as a positive control, and left to stand in the same manner. After standing, the amount of AGEs was measured by ELISA using an anti-AGE antibody (manufactured by Transgenic). From the obtained results, the AGE formation inhibition rate was calculated using the following formula. The results are shown in Tables 16-1 and 16-2. AGEs formation inhibition rate (%)={(BC) / (BA)}×100 In the above formula, A to C respectively represent the following. A: Absorbance of negative control at 405 nm B: Absorbance of positive control at 405 nm C: Absorbance at 405 nm when test sample is added

[0189] [Table 16-1]

[0190] [Table 16-2]

[0191] The results in Tables 16-1 and 16-2 confirm that the Peucedanum japonicum extract has an excellent inhibitory effect on the formation of AGEs.

[0192] (Test Example 17: Test of the effect of promoting the decomposition of advanced glycation end products (AGEs)) The Peucedanum japonicum extract was used as a test sample to test its AGE decomposition promoting effect according to the following test method.

[0193] 100 μL of 0.2 mol / L D(-)-ribose prepared in PBS(-) was added to a 96-well type I collagen-coated plate and left to stand at 37°C for 2 weeks to allow AGE formation. At this time, a plate containing only PBS(-) was also left to stand as a negative control. After 2 weeks of incubation, 100 μL of test sample (concentration: see Tables 17-1 and 17-2 below) prepared in PBS(-) was added to each well and incubated for another 20 days. A positive control was prepared by treating with D(-)-ribose and then adding PBS(-) instead of the test sample. The negative control was treated with PBS(-). After leaving it to stand for 20 days, the amount of AGEs was measured by ELISA using an anti-AGE antibody (manufactured by Transgenic). From the obtained results, the AGE decomposition promotion rate was calculated using the following formula. The results are shown in Tables 17-1 and 17-2. AGEs degradation promotion rate (%)={(BC) / (BA)}×100 In the above formula, A to C respectively represent the following. A: Absorbance of negative control at 405 nm B: Absorbance of positive control at 405 nm C: Absorbance at 405 nm when test sample is added

[0194] [Table 17-1]

[0195] [Table 17-2]

[0196] The results in Tables 17-1 and 17-2 confirm that the Peucedanum japonicum extract has an excellent effect of promoting the decomposition of AGEs.

[0197] (Test Example 18: Test of inhibitory effect on tumor necrosis factor (TNF-α) production) The extract of Peucedanum japonicum was used as a test sample and its inhibitory effect on TNF-α production was tested by the following test method.

[0198] Mouse macrophage cells (RAW264.7) were cultured in Dulbecco's MEM containing 10% FBS, and then the cells were collected using a cell scraper. The collected cells were collected at a concentration of 1.0 × 10 6 After diluting with Dulbecco's MEM containing 10% FBS to a concentration of 100 cells / mL, 100 μL was seeded per well of a 96-well plate and cultured for 4 hours. After incubation, 100 μL of the test sample (see Tables 18-1 and 18-2 below for concentrations) dissolved in 10% FBS-containing Dulbecco's MEM containing 0.5% DMSO was added to each well, and 100 μL of lipopolysaccharide (LPS, E. coli 0111;B4, DIFCO) dissolved in 10% FBS-containing Dulbecco's MEM at a final concentration of 1 μg / mL was added and incubated for 24 hours. A control was prepared by treating the same way except that the test sample was not added. After the culture was completed, the amount of TNF-α in the culture supernatant of each well was measured by sandwich ELISA. From the obtained results, the TNF-α production inhibition rate was calculated using the following formula: The results are shown in Tables 18-1 and 18-2. TNF-α production suppression rate (%)=(BA) / B×100 In the above formula, A and B respectively represent the following. A: Amount of TNF-α when test sample was added B: Amount of TNF-α without addition of test sample

[0199] [Table 18-1]

[0200] [Table 18-2]

[0201] The results in Tables 18-1 and 18-2 confirm that the Peucedanum extract has an excellent inhibitory effect on TNF-α production.

[0202] (Test Example 19: Hexosaminidase release inhibitory activity test) The Peucedanum japonicum extract was used as a test sample and its inhibitory effect on hexosaminidase release was tested by the following test method.

[0203] Rat basophilic leukemia cells (RBL-2H3) were cultured in S-MEM containing 15% FBS, and then harvested by trypsinization. The harvested cells were collected at a concentration of 4.0 × 10 5 The cells were diluted with 15% FBS-containing S-MEM to a concentration of 100 cells / mL, and DNP-specific IgE was added to a final concentration of 0.5 μL / mL. 100 μL was then seeded per well of a 96-well plate and cultured overnight. After incubation, the medium was removed and the cells were washed twice with 100 μL of Silagahanian buffer. Next, 30 μL of the same buffer and 10 μL of a test sample prepared in the same buffer (see Tables 19-1 and 19-2 below for concentrations) were added, and the cells were allowed to stand at 37°C for 10 minutes. As a control, the same procedure was performed using 40 μL of Silagahanian buffer without the test sample. Thereafter, 10 μL of a 100 ng / mL DNP-BSA solution was added, and the mixture was left to stand at 37° C. for 15 minutes to release hexosaminidase, which was then stopped by placing the 96-well plate on ice. 10 μL of the cell supernatant from each well and 10 μL of 1 mmol / L p-NAG (p-nitrophenyl-N-acetyl-β-D-glucosaminide) solution were added to a new 96-well plate and reacted at 37° C. for 1 hour. After the reaction, 250 μL of 0.1 mol / L Na2CO3 / NaHCO3 was added to each well, and the absorbance was measured at wavelengths of 415 nm and 650 nm. As a blank test, the absorbance of a mixture of 10 μL of cell supernatant and 250 μL of 0.1 mol / L Na2CO3 / NaHCO3 was measured at wavelengths of 415 nm and 650 nm, and then corrected. From the obtained results, the hexosaminidase release inhibition rate was calculated using the following formula: The results are shown in Tables 19-1 and 19-2. Hexosaminidase release inhibition rate (%) = {1-(BC) / A} x 100 In the above formula, A to C respectively represent the following. A: Absorbance at wavelengths of 415 nm and 650 nm without the addition of test sample B: Absorbance at wavelengths of 415 nm and 650 nm when test sample is added C: Absorbance at wavelengths of 415 nm and 650 nm with test sample and without p-NAG

[0204] [Table 19-1]

[0205] [Table 19-2]

[0206] The results in Tables 19-1 and 19-2 confirm that the Peucedanum extract has an excellent inhibitory effect on hexosaminidase release.

[0207] (Test Example 20: PGE2 production inhibitory effect test in mouse macrophages (COX-2 activity inhibitory effect test)) The Peucedanum japonicum extract was used as a test sample and its inhibitory effect on prostaglandin E2 (PGE2) production was tested by the following test method.

[0208] Mouse macrophage cells (RAW264.7) were cultured in Dulbecco's MEM containing 10% FBS, and then collected using a cell scraper. The collected cells were collected at a concentration of 2.0 × 10 5 After diluting with Dulbecco's MEM containing 10% FBS to a concentration of 100 cells / mL, 100 μL was seeded per well of a 96-well plate and cultured for 18 hours. After incubation, the medium was replaced with 500 μmol / L aspirin-containing medium to inactivate pre-existing cyclooxygenase-1 (COX-1) and low-expressing cyclooxygenase-2 (COX-2) by acetylation. The cells were then incubated for 4 hours. After washing three times with PBS(-), 100 μL of test sample (concentration: see Tables 20-1 and 20-2 below) dissolved in 10% FBS-containing Dulbecco's MEM containing 0.5% DMSO was added to each well. Then, 100 μL of lipopolysaccharide (LPS, E. coli 0111;B4, DIFCO) dissolved in 10% FBS-containing Dulbecco's MEM containing 10% FBS at a final concentration of 1 μg / mL was added and incubated for 16 hours. As a control, cells were incubated in the same manner using 10% FBS-containing Dulbecco's MEM containing 0.5% DMSO without the test sample. After the culture was completed, the amount of prostaglandin E2 in the culture supernatant of each well was quantified using a PGE2 EIA Kit (Cayman Chemical). From the obtained results, the PGE2 production inhibition rate was calculated using the following formula: The results are shown in Tables 20-1 and 20-2. PGE2 production suppression rate (%)={1-(AC) / (BC)}×100 In the above formula, A to C respectively represent the following. A: Amount of prostaglandin E2 after addition of test sample and stimulation with LPS B: Amount of prostaglandin E2 when stimulated with LPS without adding test sample C: Amount of prostaglandin E2 without test sample or LPS stimulation

[0209] [Table 20-1]

[0210] [Table 20-2]

[0211] The results in Tables 20-1 and 20-2 confirm that the Peucedanum extract has an excellent inhibitory effect on PGE2 production (COX-2 activity inhibitory effect).

[0212] (Test Example 21: Test of dermal papilla cell proliferation promoting activity) The Peucedanum japonicum extract was used as a test sample and tested for its hair papilla cell proliferation promoting effect by the following test method.

[0213] Normal human hair dermal papilla cells were cultured in a dermal papilla cell growth medium (manufactured by Toyobo Co., Ltd.) containing 1% FCS and growth additives, and then the cells were harvested by trypsinization. The harvested cells were diluted to 1.0 × 10 in Dulbecco's MEM containing 10% FBS. 4 After dilution to a concentration of 100 cells / mL, 200 μL was seeded per well on a collagen-coated 96-well plate and cultured for 3 days. After incubation, the medium was removed, and 200 μL of test sample dissolved in serum-free DMEM (see Tables 21-1 and 21-2 below for concentrations) was added to each well and incubated for an additional 4 days. As a control, serum-free DMEM without the test sample was used and incubated in the same manner. The proliferation-promoting activity of dermal papilla cells was measured using the MTT assay. Specifically, after incubation, the medium was removed, and 100 μL of MTT dissolved in serum-free DMEM at a final concentration of 0.4 mg / mL was added to each well. After incubation for 2 hours, the blue formazan produced within the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used to determine the amount of blue formazan produced. From the obtained results, the hair papilla cell proliferation promotion rate was calculated using the following formula. The results are shown in Tables 21-1 and 21-2. Hair papilla cell proliferation promotion rate (%) = A / B x 100 In the above formula, A and B respectively represent the following. A: Amount of blue formazan produced in cells when test sample was added B: Amount of blue formazan produced in cells without the addition of test sample

[0214] [Table 21-1]

[0215] [Table 21-2]

[0216] The results in Tables 21-1 and 21-2 confirm that the Peucedanum extract has an excellent effect of promoting the proliferation of hair follicle papilla cells.

[0217] (Test Example 22: SOD-like activity test) The Peucedanum japonicum extract was used as a test sample and its superoxide dismutase (SOD)-like activity was tested by the following test method.

[0218] A test tube was charged with 2.4 mL of 0.05 mol / L sodium carbonate buffer (pH 10.2), 0.1 mL of 3 mmol / L xanthine, 0.1 mL of 3 mmol / L EDTA, 0.1 mL of 1.5 mg / mL bovine serum albumin, and 0.1 mL of 0.75 mmol / L nitroblue tetrazolium. 0.1 mL of test sample solution (final concentrations, see Tables 22-1 and 22-2 below) was added and the mixture was incubated at 25°C for 10 minutes. Next, 0.1 mL of xanthine oxidase solution was added, and the mixture was stirred rapidly. The mixture was incubated at 25°C for 20 minutes. The reaction was then terminated by adding 0.1 mL of 6 mmol / L copper chloride, and the absorbance at 560 nm was measured. A control solution was prepared by the same procedure except that only the solvent (control solution) was added without the test sample. From the obtained results, the superoxide scavenging rate was calculated using the following formula: The results are shown in Tables 22-1 and 22-2. Superoxide scavenging rate (%) = {1-(AB) / (CD)} x 100 In the above formula, A to D respectively represent the following. A: Absorbance at 560 nm when test sample and enzyme solution are added B: Absorbance at 560 nm with test sample added and without enzyme solution added C: Absorbance at 560 nm when no test sample was added (control) and when enzyme solution was added D: Absorbance at 560 nm without adding test sample (control) or enzyme solution

[0219] [Table 22-1]

[0220] [Table 22-2]

[0221] The results in Tables 22-1 and 22-2 confirm that the Peucedanum japonicum extract has excellent superoxide scavenging activity.

[0222] (Test Example 23: DPPH radical scavenging activity test) The Peucedanum japonicum extract was used as a test sample, and the radical scavenging activity was tested using diphenyl-p-picrylhydrazyl (DPPH), a very stable radical, according to the following test method.

[0223] 3 mL of test sample solution (see Tables 23-1 and 23-2 below for concentrations) was added to 3 mL of 150 μmol / L DPPH ethanol solution, the container was sealed, shaken, and left to stand for 30 minutes. After standing, the absorbance at a wavelength of 520 nm was measured. As a control, the same procedure was performed using only the solvent (control solution) without the test sample. From the obtained results, the DPPH radical scavenging rate (sometimes referred to as the DPPH scavenging rate) was calculated using the following formula. The results are shown in Tables 23-1 and 23-2. DPPH erasure rate (%)={A-(BC)} / A×100 In the above formula, A to C respectively represent the following. A: Absorbance at 520 nm when no test sample was added (control) and when DPPH solution was added B: Absorbance at 520 nm when test sample and DPPH solution are added C: Absorbance at a wavelength of 520 nm when test sample is added and DPPH solution is not added

[0224] [Table 23-1]

[0225] [Table 23-2]

[0226] The results in Tables 23-1 and 23-2 confirm that Peucedanum japonicum extract has excellent radical scavenging activity.

[0227] (Combination example 1) Tablets having the following composition were prepared by a conventional method. Peucedanum extract 5.0mg (Extract produced in Production Example 3) Dolomite 83.4mg (Contains 20% calcium and 10% magnesium) Casein phosphopeptide 16.7mg Vitamin C 33.4mg Maltitol 136.8mg Collagen 12.7mg Sucrose fatty acid esters 12.0mg

[0228] (Combination example 2) An oral liquid preparation having the following composition was prepared by a conventional method. <Composition in 1 ampoule (100 mL)> Peucedanum officinalis extract 0.3% by mass (Extract produced in Production Example 5) Sorbitol 12.0% by mass Sodium benzoate 0.1% by mass · Fragrance 1.0% by mass Calcium sulfate 0.5% by mass · Purified water remainder

[0229] (Combination example 3) An emulsion having the following composition was prepared by a conventional method. Peucedanum officinalis extract (extract produced in Production Example 1) 0.01g Jojoba oil 4.00g 1,3-butylene glycol 3.00g Arbutin 3.00g Polyoxyethylene cetyl ether (20E.O.) 2.50g 2.00g olive oil 2.00g squalane 2.00g of cetanol Glyceryl monostearate 2.00g Polyoxyethylene sorbitan oleate (20E.O.) 2.00g Methyl parahydroxybenzoate 0.15g Stearyl glycyrrhizinate 0.10g Phellodendron Bark Extract 0.10g Dipotassium glycyrrhizinate 0.10g Ginkgo biloba extract 0.10g 0.10g conchiolin Phellodendron bark extract 0.10g Chamomile extract 0.10g · Fragrance 0.05g Purified water, balance (total amount 100g)

[0230] (Combination example 4) A cream having the following composition was prepared by a conventional method. Peucedanum officinalis extract (extract produced in Production Example 2) 0.05g Sophora root extract 0.1g Scutellaria root extract 0.1g Liquid paraffin 5.0g 4.0g white beeswax Squalane 10.0g Cetyl alcohol 3.0g 2.0g lanolin Stearic acid 1.0g Polyoxyethylene sorbitan oleate (20E.O.) 1.5g Glyceryl monostearate 3.0g Oil-soluble licorice extract 0.1g 1,3-butylene glycol 6.0g Methyl parahydroxybenzoate 1.5g · Fragrance 0.1g Purified water, balance (total amount 100g)

[0231] (Combination example 5) A cosmetic essence having the following composition was prepared by a conventional method. Peucedanum officinalis extract (extract produced in Production Example 4) 0.01g Chamomile extract 0.1g 0.3g xanthan gum Hydroxyethylcellulose 0.1g Carboxyvinyl polymer 0.1g 1,3-butylene glycol 4.0g Dipotassium glycyrrhizinate 0.1g 2.0g glycerin 0.25g potassium hydroxide · Fragrance 0.01g Preservative (methyl parahydroxybenzoate) 0.15g 2.0g ethanol Purified water, balance (total amount 100g)

[0232] (Combination example 6) A hair tonic having the following composition was prepared by a conventional method. Peucedanum officinalis extract (extract produced in Production Example 6) 0.4g Tocopherol acetate (appropriate amount) Cephalatin 0.002g Isopropylmethylphenol 0.1g Sodium hyaluronate 0.15g 15.0g glycerin 15.0g ethanol · Appropriate amount of fragrance Chelating agent (sodium edetate) appropriate amount Preservative (hinokitiol) appropriate amount Solubilizer (polyoxyethylene cetyl ether) appropriate amount Purified water, balance (total amount 100g)

[0233] (Combination example 7) A shampoo having the following composition was prepared by a conventional method. Peucedanum officinalis extract (extract produced in Production Example 3) 0.5g Marjoram extract 1.0g Plum fruit extract 0.2g Sodium coconut oil fatty acid methyl taurate 10.0g Coconut oil fatty acid amidopropyl betaine 10.0g Sodium polyoxyethylene alkyl ether sulfate 20.0g Coconut oil fatty acid diethanolamide 4.0g Propylene glycol 2.0g · Appropriate amount of fragrance Purified water, balance (total amount 100g)

Claims

1. An immunostimulant characterized by containing an extract of Peucedanum japonicum.

2. The immunostimulant according to claim 1, which has an effect of promoting TNF-α production.

3. An immunostimulating composition comprising the immunostimulant according to claim 1 or 2.

4. A TNF-α production promoter characterized by containing an extract of Peucedanum japonicum.

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