Heat shock protein gene expression regulator, pharmaceutical product, cosmetic, and method for producing heat shock protein gene expression regulator

Abstract

The present invention provides a heat shock protein gene expression regulator having the genus Lactobacillus derived from Artemisia argyi or Angelica keiskei.

Description

Technical Field

[0001] The present invention relates to a heat shock protein gene expression regulator, a medicine, a cosmetic and a method for producing the heat shock protein gene expression regulator. Background Art

[0002] In recent years, with the development of research on the structure of human skin and its metabolic mechanism, the causes and mechanisms of changes such as wrinkles, fine lines, spots and sagging in human skin with age are becoming increasingly clear. The skin is composed of a thin epidermis (epithelial tissue) on the outside and a thick dermis (connective tissue) underneath. As the outermost layer of the body, the epidermis protects the organism from external damage and prevents the internal moisture and nutrients from leaking to the outside. The dermis is a connective tissue with a structure that mainly includes fibroblasts, collagen fibers (collagen), elastic fibers (elastin) and proteoglycans that expand in a three-dimensional shape, and is responsible for giving the skin strength, stretchability and elasticity. If the amount of sebum and water in the skin decreases with age, the moisturizing power of the stratum corneum on the surface of the skin will be lost, and fine lines and rough skin caused by dryness, etc. are likely to occur. In response to this, it has been proposed to use fermented products to improve skin function (for example, refer to patent documents 1 to 7). In addition, it has been proposed that the expression of heat shock proteins can be induced by administering an extract of Arnica montana or the like, thereby bringing about a whitening effect (for example, see Patent Document 8).

[0003] Prior art literature

[0004] Patent Literature

[0005] Patent Document 1: International Publication No. 2018 / 123828

[0006] Patent Document 2: Japanese Patent No. 5468183

[0007] Patent Document 3: Japanese Patent Application Publication No. 2015-156832

[0008] Patent Document 4: Japanese Patent No. 5467106

[0009] Patent Document 5: Japanese Patent No. 4990297

[0010] Patent Document 6: Japanese Patent Application Publication No. 2009-249366

[0011] Patent Document 7: Japanese Patent Application Publication No. 2009-249365

[0012] Patent Document 8: Japanese Patent No. 5697879 Summary of the invention

[0013] The object of the present invention is to provide a heat shock protein gene expression regulator, a pharmaceutical product, a cosmetic product, and a method for producing a heat shock protein gene expression regulator that uses a natural ingredient as an active ingredient and exhibits a wide range of effects.

[0014] A heat shock protein gene expression regulator of one aspect of the present invention comprises Lactobacillus genus from mugwort or Angelica keiskei. A pharmaceutical product of one aspect of the present invention comprises Lactobacillus genus from mugwort or Angelica keiskei. A cosmetic product of one aspect of the present invention comprises Lactobacillus genus from mugwort or Angelica keiskei. A method for producing a heat shock protein gene expression regulator of one aspect of the present invention comprises obtaining Lactobacillus genus from mugwort or Angelica keiskei.

[0015] According to the present invention, a heat shock protein gene expression regulator, a pharmaceutical product, a cosmetic product, and a method for producing a heat shock protein gene expression regulator which contains a natural ingredient as an active ingredient and exhibits a wide range of effects can be provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 These are a graph and a table showing the analysis results of the bacteria contained in the fermentation liquid of Example 1.

[0017] Figure 2 These are a graph and a table showing the analysis results of the bacteria contained in the fermentation liquid of Example 2.

[0018] Figure 3 This is a table showing the analysis results of gene expression in Example 4.

[0019] Figure 4 This is a table showing the analysis results of gene expression in Example 4.

[0020] Figure 5 This is a table showing the analysis results of gene expression in Example 4.

[0021] Figure 6 This is a table showing the analysis results of gene expression in Example 4.

[0022] Figure 7 This is a table showing the analysis results of gene expression in Example 4.

[0023] Figure 8 This is a table showing the analysis results of gene expression in Example 4.

[0024] Fig. 9 These are photographs of people after administration of the fermentation liquid of Example 5.

[0025] Fig.10 This is a graph showing the antibacterial effect of the fermentation liquid of Example 6.

[0026] Fig.11 This is a graph showing the antifungal effect of the fermentation liquid of Example 7.

[0027] Fig.12 This is a graph showing the antiviral effect of the fermentation liquid of Example 8.

[0028] Fig.13 This is a graph showing the antiviral effect of the fermentation liquid of Example 8.

[0029] Fig.14 This is a graph showing the anti-Trichophyton effect of the fermentation liquid of Example 9.

[0030] Fig.15 This is a table showing the production amount of type I collagen in Example 10.

[0031] Fig.16 This is a table showing the production amount of HSP47 in Example 10.

[0032] Fig.17 This is a microscope image of cells stained with SA-β-Gal in Example 11.

[0033] Fig.18 This is a microscope image of cells stained with SA-β-Gal in Example 11.

[0034] Fig.19 This is a table showing the degree of SA-β-Gal staining in Example 11.

[0035] Fig. 20 This is a table showing the cell survival rate of Example 12.

[0036] Fig.21 This is a table showing the production amount of HSP70 in Example 12. DETAILED DESCRIPTION

[0037] The following is a detailed description of the embodiments of the present invention. It should be noted that the embodiments shown below are examples of devices and methods for implementing the technical ideas of the present invention, and the technical ideas of the present invention do not limit the combination of components to the following contents. The technical ideas of the present invention can be modified in various ways within the scope of the claims.

[0038] The heat shock protein (HSP) gene expression regulator of the embodiment has the genus Lactobacillus from mugwort or Angelica keiskei. The heat shock protein gene expression regulator of the embodiment particularly promotes the expression of the heat shock protein (HSP) 70 gene that improves the anti-aging function. The genus Lactobacillus is a type of lactic acid bacteria and is a Gram-positive facultative anaerobic bacterium. The genus Lactobacillus ferments sugar to produce lactic acid. The genus Lactobacillus also inhabits animals including humans, but the genus Lactobacillus of the embodiment is the genus Lactobacillus from mugwort or Angelica keiskei.

[0039] For example, the Lactobacillus of the embodiment is extracted by fermenting mugwort or ashitaba. The heat shock protein gene expression regulator of the embodiment may further include a fermentation liquid of mugwort or ashitaba.

[0040] Examples of Lactobacillus derived from Artemisia include parafarraginis, parabuchneri, buchneri, and harbinensis. Examples of Lactobacillus derived from Angelica keiskei include vini and nagelii. The heat shock protein gene expression regulator of the embodiment may include a plurality of species of Lactobacillus.

[0041] The heat shock protein gene expression regulator of the embodiment promotes the expression of heat shock protein genes. As examples of heat shock protein genes, the HSPA1A gene encoding HSP70 and the HSPB1 gene encoding HSP27 can be cited. HSP70 has whitening promotion function, anti-stain function, anti-wrinkle function, anti-stress function, anti-cell death function, anti-inflammatory function, cell protection function, gastric mucosa protection function and anti-DNA damage function. HSP27 has anti-stress function and wound healing function. The heat shock protein gene expression regulator of the embodiment has the Lactobacillus (Lactobacillus llus) genus from Angelica keiskei, which promotes the production of HSP47. HSP47 is a molecular chaperone involved in the biosynthesis of collagen. Therefore, the heat shock protein gene expression regulator of the embodiment can improve, for example, anti-stress function, anti-cell death function, anti-inflammatory function, cell protection function, gastric mucosa protection function, anti-DNA damage function and wound healing function. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting heat shock protein expression, improving whitening promoting function, improving anti-spot function, improving anti-wrinkle function, improving anti-stress function, improving anti-cell death function, improving anti-inflammatory function, improving cell protection function, improving gastric mucosa protection function, improving anti-DNA damage function and wound healing. It should be noted that wrinkles include fine lines.

[0042] The heat shock protein gene expression regulator of the embodiment also regulates the expression of genes other than the heat shock protein gene. For example, the heat shock protein gene expression regulator of the embodiment inhibits the expression of the acid ceramidase (ceramidase) gene. As an example of an acid ceramidase gene, the ASAH1 gene can be cited. In order to inhibit the expression of acid ceramidase, the heat shock protein gene expression regulator of the embodiment can, for example, promote ceramide biosynthesis and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from ceramidase expression inhibition, ceramide biosynthesis promotion and barrier function improvement.

[0043] The heat shock protein gene expression regulator of the embodiment promotes the expression of lipid synthase genes. As an example of a lipid synthase gene, the DGAT1 gene can be cited. DGAT1 (diacylglycerol O-acyltransferase 1) synthesizes neutral fat (TAG) from diacylglycerol (DAG). The barrier function of the skin of DGAT1-deficient mice is abnormal. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, promote lipid synthesis and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting lipid synthesis and improving barrier function.

[0044] The heat shock protein gene expression regulator of the embodiment promotes the expression of lysosomal hydrolase genes. Lysosomes decompose sugars and glycolipids in cells. In order to exert the function of lysosomes, lysosomal hydrolases are required. As an example of a lysosomal hydrolase gene, an acid β-glucosidase (GBA) gene can be cited. In patients with lysosomal diseases, the activity of GBA is reduced or absent. In addition, the activation of GBA improves the barrier function of the skin. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, prevent and treat lysosomal diseases and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of lysosomal hydrolases, improving lysosomal function, preventing lysosomal diseases, treating lysosomal diseases and improving barrier function.

[0045] The heat shock protein gene expression regulator of the embodiment promotes the expression of tight junction biosynthesis factor genes. As examples of tight junction biosynthesis factor genes, occludin 1 (CLDN1) gene and occludin (OCLN) gene can be cited. Ocludin and occludin are components of tight junctions. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, promote the biosynthesis of tight junctions and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of tight junction biosynthesis factors, promoting the expression of occludin, promoting the expression of occludin, promoting the biosynthesis of tight junctions and improving the barrier function.

[0046] The heat shock protein gene expression regulator of the embodiment promotes the expression of the intercellular adhesion factor gene. As examples of intercellular adhesion factor genes, integrin α2 (ITGA2) gene, E-cadherin (CDH1) gene and hyaluronic acid receptor (CD44) gene can be cited. Integrin, cadherin and CD44 contribute to intercellular adhesion. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, promote the biosynthesis of intercellular adhesion factors and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the promotion of intercellular adhesion factor biosynthesis, promotion of integrin expression, promotion of cadherin expression, promotion of hyaluronic acid receptor expression and improvement of barrier function.

[0047] The heat shock protein gene expression regulator of the embodiment promotes the expression of temperature-sensitive TRP (transient receptor potential) channel genes. As an example of TRP channel genes, the TRPV3 gene can be cited. TRPV3 is involved in the proliferation and differentiation of keratinocytes that maintain the barrier function of the epidermis. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, promote the biosynthesis of keratinocytes and improve the barrier function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting TRP channel expression, promoting keratinocyte biosynthesis and improving barrier function.

[0048] The heat shock protein gene expression regulator of the embodiment promotes the expression of antimicrobial molecule genes. As an example of antimicrobial molecule genes, the Toll-like receptor 2 (TLR2) gene can be cited. Toll-like receptors have the function of sensing microorganisms and activating immunity. In addition, as examples of antimicrobial molecule genes, the DEFB1 gene, the DEFB4A gene, and the DEFB103A gene can be cited. The DEFB1 gene, the DEFB4A gene, and the DEFB103A gene encode beta defensins such as DEFB1, DEFB2, and DEFB103, which are antimicrobial peptides. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the antimicrobial function. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic, and / or food for at least one application selected from the group consisting of promoting the expression of Toll-like receptors, promoting the expression of beta defensins, and improving the antimicrobial function.

[0049] The heat shock protein gene expression regulator of the embodiment promotes the expression of sirtuin genes. As an example of sirtuin genes, the SIRT4 gene can be cited. The activity of the sirtuin gene improves the anti-aging function. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the anti-aging function. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting sirtuin expression and improving anti-aging function.

[0050] The heat shock protein gene expression regulator of the embodiment promotes the expression of the telomerase gene. As examples of telomerase genes, TERT gene and TERC gene can be cited. If the activity of telomerase is promoted, the life span of the cell becomes longer. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the anti-aging function. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting telomerase expression and improving anti-aging function.

[0051] The heat shock protein gene expression regulator of the embodiment promotes the expression of energy metabolism factor genes. As an example of an energy metabolism factor gene, the PPARGC1A gene can be cited. The PPARGC1A gene encodes PGC-1 (peroxisome proliferator-activated receptor gamma coactivator 1). PGC-1 promotes energy metabolism. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve energy metabolism function. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of energy metabolism factors, promoting the expression of PGC-1 and improving the energy metabolism function.

[0052] The heat shock protein gene expression regulator of the embodiment promotes the expression of hyaluronan biosynthesis factor genes. Examples of hyaluronan biosynthesis factor genes include hyaluronan synthase 1 (HAS1) gene, hyaluronan synthase 2 (HAS2) gene, and hyaluronan synthase 3 (HAS3) gene.

[0053] In addition, the heat shock protein gene expression regulator of the embodiment inhibits the expression of hyaluronan decomposition factor gene. Examples of hyaluronan decomposition factor genes include hyaluronan decomposition enzyme 2 (HYAL2) gene and cell migration-inducing and hyaluronan-binding protein (CEMIP, KIAA1199) gene.

[0054] Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the hyaluronic acid biosynthesis function. Hyaluronic acid is effective for the prevention and treatment of osteoarthritis, the improvement of moisturizing function, the elimination of relaxation and the reduction of wrinkles. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, prevent and treat osteoarthritis, improve moisturizing function, reduce relaxation and wrinkles. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from hyaluronan synthase expression promotion, hyaluronan decomposition factor expression inhibition, hyaluronic acid biosynthesis promotion, osteoarthritis prevention, osteoarthritis treatment, moisturizing function improvement, anti-relaxation function improvement and anti-wrinkle function improvement.

[0055] The heat shock protein gene expression regulator of the embodiment promotes the expression of the basement membrane biosynthesis factor gene. Examples of basement membrane biosynthesis factor genes include laminin α1 (LAMA1) gene, laminin α5 (LAMA5) gene, type IV collagen α1 (COL4A1) gene, and type VII collagen α1 (COL7A1). The heat shock protein gene expression regulator of the embodiment can, for example, promote the biosynthesis of laminin and collagen, and promote the biosynthesis of basement membrane. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic, and / or food for at least one application selected from the group consisting of promoting the expression of basement membrane biosynthesis factors, promoting the expression of laminin, promoting the expression of collagen, and promoting the biosynthesis of basement membrane.

[0056] The heat shock protein gene expression regulator of the embodiment promotes the expression of type I collagen. Type I collagen imparts elasticity to bones. Type I collagen imparts strength to skin. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic, and / or food for promoting the expression of type I collagen.

[0057] The heat shock protein gene expression regulator of the embodiment promotes the expression of the matrix metalloproteinase inhibitor gene. As an example of the matrix metalloproteinase inhibitor gene, the TIMP1 gene can be cited. The matrix metalloproteinase inhibitor inhibits the matrix metalloproteinase and inhibits the decomposition of the extracellular matrix. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, inhibit the decomposition of the extracellular matrix. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of the matrix metalloproteinase inhibitor and inhibiting the decomposition of the extracellular matrix.

[0058] The heat shock protein gene expression regulator of the embodiment promotes the expression of antioxidant factor genes. As examples of antioxidant factor genes, superoxide dismutase 3 (SOD3) gene, catalase (CAT) gene, glutathione reductase (GSR) gene, glutathione peroxidase 1 (GPX1) gene, metallothionein 1F (MT1F) gene can be cited. SOD is an enzyme that decomposes active oxygen. CAT is an enzyme that decomposes hydrogen peroxide. GSR is an enzyme that reduces oxidized glutathione. GPX1 is an enzyme that decomposes hydrogen peroxide. MT1F is an antioxidant protein. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the antioxidant function. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of antioxidant factor genes, promoting the expression of SOD, promoting the expression of CAT, promoting the expression of GSR, promoting the expression of GPX1, promoting the expression of MT1F and improving the antioxidant function.

[0059] The heat shock protein gene expression regulator of the embodiment promotes the expression of the interleukin 1 receptor antagonist (IL1RN) gene. If the interleukin 1 receptor antagonist is missing, interleukin 1 receptor antagonist deficiency (DIRA) is produced. Therefore, the heat shock protein gene expression regulator of the embodiment can prevent and treat DIRA. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of interleukin 1 receptor antagonist, preventing DIRA and treating DIRA.

[0060] The heat shock protein gene expression regulator of the embodiment promotes the expression of the semaphorin gene. As an example of the semaphorin gene, the SEMA3A gene can be cited. The semaphorin is an exclusionary guidance factor that exhibits an exclusionary effect on the elongation of nerve axons. The semaphorin inhibits angiogenesis. The semaphorin controls bone weight. In addition, the semaphorin inhibits itching. The semaphorin has preventive and therapeutic effects on atopic dermatitis. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, repulsively guide the elongation of nerve axons, inhibit angiogenesis, control bone weight, inhibit itching, or prevent and treat atopic dermatitis. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of semaphorins, inhibiting the elongation of nerve axons, inhibiting angiogenesis, controlling bone weight, inhibiting itching, preventing atopic dermatitis and treating atopic dermatitis.

[0061] The heat shock protein gene expression regulator of the embodiment promotes the expression of nerve growth factor (NGF) gene and nerve growth factor receptor (NGFR). Nerve growth factor promotes the growth and maintenance of nerves, promotes the functional recovery of cranial nerves, and is effective for the prevention and treatment of Alzheimer's disease and dementia. Nerve growth factor acts through nerve growth factor receptors. Therefore, the heat shock protein gene expression regulator of the embodiment, for example, can promote the expression of nerve growth factor, promote the growth and maintenance of nerves, promote the functional recovery of cranial nerves, prevent and treat Alzheimer's disease and dementia. The heat shock protein gene expression regulator of the embodiment, for example, can be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting nerve growth factor expression, promoting nerve growth and maintenance, promoting cranial nerve functional recovery, preventing Alzheimer's disease, treating Alzheimer's disease, preventing dementia and treating dementia.

[0062] The heat shock protein gene expression regulator of the embodiment promotes the expression of the inhibitor gene of nuclear factor κB (NFκB). As an example of the inhibitor gene of NFκB, the NFKBIA gene can be cited. The inhibitor of NFκB such as IκBα inhibits the activity of NFκB to prevent and treat cancer. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, prevent and treat cancer. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of NFκB inhibitor, cancer prevention and cancer treatment.

[0063] The heat shock protein gene expression regulator of the embodiment promotes the expression of the calpain inhibitor protein (CAST) gene, which is a calpain (CAPN) inhibitor protein. In aging cells, CAPN is activated and CAST disappears. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, inhibit aging. The heat shock protein gene expression regulator of the embodiment can be used as a pharmaceutical, cosmetic and / or food for use selected from the promotion of CAPN inhibitor protein expression and the improvement of anti-aging function.

[0064] The heat shock protein gene expression regulator of the embodiment promotes the expression of the citrullinated protein activator gene. As an example of the citrullinated protein activator gene, peptidyl arginine deiminase 3 (PAD3) can be cited. PAD activates citrullinated proteins and promotes the keratinization of normal epidermis. Therefore, the heat shock protein gene expression regulator of the embodiment, for example, can promote the keratinization of normal skin. The heat shock protein gene expression regulator of the embodiment, for example, can be used as a pharmaceutical, cosmetic and / or food for use selected from the promotion of citrullinated protein activator expression and the promotion of keratinization of normal skin.

[0065] The heat shock protein gene expression regulator of the embodiment promotes the expression of the transglutaminase gene. As an example of the transglutaminase gene, the TGM1 gene can be cited. Transglutaminase increases the physical strength of the skin surface, or improves the moisturizing function. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, increase the strength of the skin surface, or improve the moisturizing function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting transglutaminase expression, improving skin surface strength, improving skin elasticity and improving skin moisturizing function.

[0066] The heat shock protein gene expression regulator of the embodiment promotes the expression of the involucrin (IVL) gene. IVL promotes the maturation of the cornified envelope and improves the moisturizing function of the skin. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, promote the maturation of the cornified envelope and improve the moisturizing function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting IVL expression, promoting maturation of the cornified envelope and improving the moisturizing function of the skin.

[0067] The heat shock protein gene expression regulator of the embodiment promotes the expression of aquaporin genes. As an example of aquaporin genes, the AQP3 gene can be cited. Aquaporins participate in the migration of keratinocytes and improve the moisturizing function of the skin. Therefore, the heat shock protein gene expression regulator of the embodiment can, for example, improve the migration function of keratinocytes and improve the moisturizing function of the skin. The heat shock protein gene expression regulator of the embodiment can, for example, be used as a pharmaceutical, cosmetic and / or food for at least one application selected from the group consisting of promoting the expression of aquaporins, improving the migration function of keratinocytes and improving the moisturizing function of the skin.

[0068] In addition, the heat shock protein gene expression regulator of the embodiment also has the function of reducing fungi (molds). The heat shock protein gene expression regulator of the embodiment, for example, reduces fungi by more than 80%, more than 85%, more than 90%, or more than 95% within 24 hours. Examples of fungi include, but are not limited to, white ringworm, candida, cryptococcus, and aspergillus. The heat shock protein gene expression regulator of the embodiment also has the therapeutic effect of fungal diseases. Examples of fungal diseases include, but are not limited to, white ringworm, candida, cryptococcosis, and aspergillosis.

[0069] The heat shock protein gene expression regulator of the embodiment also has the function of reducing Gram-negative bacteria and Gram-positive bacteria. The heat shock protein gene expression regulator of the embodiment, for example, reduces Gram-negative bacteria and Gram-positive bacteria by more than 80%, more than 85%, more than 90% or more than 95% within 24 hours. As Gram-negative bacteria, Escherichia coli, Salmonella, Vibrio parahaemolyticus, Klebsiella pneumoniae and Pseudomonas aeruginosa can be cited, but are not limited to these. As Gram-positive bacteria, methicillin-resistant Staphylococcus aureus (MRSA), spore-forming Bacillus cereus and Bacillus subtilis can be cited, but are not limited to these.

[0070] The heat shock protein gene expression regulator of the embodiment also has the function of reducing viruses. The heat shock protein gene expression regulator of the embodiment, for example, reduces viruses by 80% or more, 85% or more, 90% or more, or 95% or more within 24 hours. Viruses include enveloped viruses, which are viruses with envelopes, and non-enveloped viruses, which are viruses without envelopes. In addition, viruses include DNA viruses and RNA viruses.

[0071] Examples of DNA viruses having an envelope include human herpes virus, vaccinia virus, and hepatitis B virus, but are not limited to these.

[0072] Examples of RNA viruses having an envelope include, but are not limited to, influenza virus, SARS coronavirus, RS virus, mumps virus, Lassa virus, dengue virus, rubella virus, human immunodeficiency virus, measles virus, hepatitis C virus, Ebola virus, yellow fever virus, and Japanese encephalitis virus.

[0073] Examples of DNA viruses that do not have an envelope include adenovirus, B19 virus, papovavirus, and human papillomavirus, but are not limited to these.

[0074] Examples of RNA viruses not having an envelope include norovirus, poliovirus, echovirus, hepatitis A virus, hepatitis E virus, rhinovirus, astrovirus, rotavirus, coxsackievirus, enterovirus, and sapovirus, but are not limited thereto.

[0075] The heat shock protein gene expression regulator of the embodiment contains an effective amount of Lactobacillus. The heat shock protein gene expression regulator of the embodiment may contain Lactobacillus in a solvent such as water, or may contain Lactobacillus in a fermentation liquid of a plant. As plants for obtaining the fermentation liquid, mugwort and ashitaba can be mentioned as in the case of Lactobacillus.

[0076] Effective amount refers to the amount required for the gene expression regulation effect, and can be appropriately determined according to the gene as the object. Generally speaking, when the concentration of Lactobacillus is high and the solvent contains the fermented liquid of the plant, the gene expression regulation effect is played, but for example, when the target gene is the GBA gene, the concentration of Lactobacillus is low, and the gene expression regulation effect is played. In addition, for example, when the target gene is the TLR2 gene, the solvent does not contain the fermented liquid of the plant, and the gene expression regulation effect is played.

[0077] The Lactobacillus contained in the heat shock protein gene expression regulator of the embodiment can be live bacteria or dead bacteria treated by heating, for example. Therefore, the heat shock protein gene expression regulator of the embodiment can contain dead bacteria of the Lactobacillus. The Lactobacillus can be a dried cell. The dead bacteria or dried cell of the Lactobacillus also play a gene expression regulation effect and an antibacterial and antiviral effect. In addition, the dead bacteria or dried cell of the Lactobacillus is easy to transport and store for a long time.

[0078] The heat shock protein gene expression regulator of the embodiment may be, for example, a liquid, a cream, an ointment, a plaster, a gel, a wax, and a spray.

[0079] The heat shock protein gene expression regulator of the embodiment may be, for example, a cosmetic for whole skin. Examples of cosmetics for whole skin include lotions, beauty serums, and facial masks. The heat shock protein gene expression regulator of the embodiment may be, for example, a protective cosmetic. Examples of protective cosmetics include protective lotions and protective creams. The heat shock protein gene expression regulator of the embodiment may be, for example, a base cosmetic. Examples of base cosmetics include foundation, pressed powder, and primer. The heat shock protein gene expression regulator of the embodiment may be, for example, a point make-up cosmetic. Examples of point make-up cosmetics include lipstick, eye makeup, blush, and nail polish.

[0080] In addition, the heat shock protein gene expression regulator of the embodiment is provided in the form of a skin external medicine such as a disinfectant, a coating treatment medicine, an eye drop, or an oral medicine. The heat shock protein gene expression regulator of the embodiment can be administered to human skin including fingers and toes, hair, oral cavity, eyeball, etc.

[0081] The heat shock protein gene expression regulator of the embodiment may contain, in addition to Lactobacillus, cosmetic and pharmaceutical ingredients such as liquid oils, solid oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, humectants, water-soluble polymers, thickeners, film-forming agents, metal ion chelating agents, lower alcohols, polyols, sugars, amino acids, organic amines, pH adjusters, skin nutrients, vitamins, antioxidants, fragrances, powders, coloring materials, and water, as appropriate depending on the purpose.

[0082] When the heat shock protein gene expression regulator of the embodiment contains an oily component, the concentration of the oily component in the heat shock protein gene expression regulator of the embodiment is not particularly limited, and is, for example, 0.1% to 90% by mass or 0.5% to 90% by mass. When the heat shock protein gene expression regulator of the embodiment contains an aqueous component, the concentration of the aqueous component in the heat shock protein gene expression regulator of the embodiment is not particularly limited, and is, for example, 0.1% to 90% by mass or 0.5% to 90% by mass. The ratio of the oily component to the aqueous component in the heat shock protein gene expression regulator of the embodiment can be appropriately set depending on whether the heat shock protein gene expression regulator of the embodiment is an oil-in-water (O / W) type or a water-in-oil (W / O) type. When the heat shock protein gene expression regulator of the embodiment contains a surfactant, the concentration of the surfactant in the heat shock protein gene expression regulator of the embodiment is not particularly limited, and is, for example, 2% to 10% by mass.

[0083] The heat shock protein gene expression regulator of the embodiment may contain an antibacterial substance or an antiviral substance as appropriate depending on the purpose, in addition to Lactobacillus.

[0084] The heat shock protein gene expression regulator of the embodiment can be produced by fermenting plants to obtain a fermentation liquid containing Lactobacillus. When fermenting plants, sugars such as salt and molasses can be added to the plants. The fermentation temperature is, for example, 30°C. The hydrogen ion index (pH) of the obtained fermentation liquid is about 4.0. The secretion of Lactobacillus can be extracted from the fermentation liquid.

[0085] The obtained fermented liquid can be heated to make the Lactobacillus contained in the fermented liquid become dead bacteria. In addition, the fermented liquid can be spray-dried to obtain a dried cell of the Lactobacillus genus. The dried cell can also be produced by freeze drying and hot air drying.

[0086] The obtained fermented liquid, Lactobacillus bacteria or dried Lactobacillus bacteria can be added to soy milk to ferment the soy milk to obtain soy milk fermented liquid. The soy milk fermented liquid also has a gene expression regulating effect.

[0087] As described above, the heat shock protein gene expression regulator and the like according to each embodiment of the present invention have the following configurations and effects consisting of any one or more combinations of the above.

[0088] The heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment comprises Lactobacillus genus from mugwort or Angelica keiskei. The heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment may further comprise fermentation liquid from mugwort or Angelica keiskei. The medicine may be an anti-wrinkle medicine or an anti-aging medicine. The cosmetic may be an anti-wrinkle cosmetic or an anti-aging cosmetic.

[0089] This embodiment includes Lactobacillus from mugwort or Angelica keiskei for regulating the expression of heat shock protein genes. This embodiment includes Lactobacillus from mugwort or Angelica keiskei for regulating the expression of heat shock protein genes and a fermentation liquid from mugwort or Angelica keiskei.

[0090] This embodiment includes the use of Lactobacillus from mugwort or Angelica keiskei for producing a heat shock protein gene expression regulator, a pharmaceutical product, or a cosmetic product. This embodiment includes the use of Lactobacillus from mugwort or Angelica keiskei and a fermentation liquid from mugwort or Angelica keiskei for producing a heat shock protein gene expression regulator, a pharmaceutical product, or a cosmetic product.

[0091] This embodiment includes a method for regulating the expression of heat shock protein genes, a method for treating or a method for beauty, comprising administering Lactobacillus from mugwort or Angelica keiskei to a human or non-human animal. This embodiment includes a method for regulating the expression of heat shock protein genes, a method for treating or a method for beauty, comprising administering Lactobacillus from mugwort or Angelica keiskei and a fermented liquid from mugwort or Angelica keiskei to a human or non-human animal. The method for treating may be a method for improving the anti-wrinkle function or a method for improving the anti-aging function. The method for beauty may be a method for improving the anti-wrinkle function or a method for improving the anti-aging function.

[0092] The heat shock protein gene may be at least one selected from the group consisting of the HSPA1A gene and the HSPB1 gene. The Lactobacillus genus derived from mugwort or Angelica keiskei of the present embodiment may promote the expression of at least one selected from the group consisting of the HSPA1A gene and the HSPB1 gene.

[0093] The Lactobacillus genus from mugwort or Angelica keiskei of this embodiment can regulate the expression of genes other than heat shock protein genes. The gene whose expression is regulated by the Lactobacillus genus from mugwort or Angelica keiskei of this embodiment can be at least one selected from the group consisting of ceramidase gene, lipid synthase gene, lysosomal hydrolase gene, tight junction biosynthesis factor gene and intercellular adhesion factor gene.

[0094] The ceramidase gene may be the ASAH1 gene. The Lactobacillus genus derived from mugwort or Angelica keiskei of the present embodiment can inhibit the expression of the ASAH1 gene.

[0095] The lipid synthase gene may be a DGAT1 gene. The Lactobacillus genus derived from Artemisia annua or Angelica keiskei of the present embodiment can promote the expression of the DGAT1 gene.

[0096] The lysosomal hydrolase gene may be a GBA gene. The Lactobacillus genus derived from Artemisia argyi or Angelica keiskei of this embodiment can promote the expression of the GBA gene.

[0097] The tight junction biosynthesis factor gene may be at least one selected from the group consisting of the CLDN1 gene and the OCLN gene. The Lactobacillus genus derived from mugwort or Angelica keiskei of the present embodiment may promote the expression of at least one selected from the group consisting of the CLDN1 gene and the OCLN gene.

[0098] The intercellular adhesion factor gene may be at least one selected from the group consisting of ITGA2 gene, CDH1 gene, and CD44 gene. The Lactobacillus genus derived from mugwort or Angelica keiskei of this embodiment may promote the expression of at least one selected from the group consisting of ITGA2 gene, CDH1 gene, and CD44 gene.

[0099] The gene regulated by the Lactobacillus from mugwort or Angelica keiskei of the present embodiment may be an antimicrobial molecule gene. The antimicrobial molecule gene may be at least one selected from the group consisting of TLR2 gene, DEFB1 gene, DEFB4A gene, and DEFB103A gene. The Lactobacillus from mugwort or Angelica keiskei of the present embodiment may promote the expression of at least one selected from the group consisting of TLR2 gene, DEFB1 gene, DEFB4A gene, and DEFB103A gene.

[0100] The gene whose expression is regulated by Lactobacillus derived from Artemisia argyi or Angelica keiskei of the present embodiment may be at least one selected from sirtuin gene and telomerase gene.

[0101] The sirtuin gene is the SIRT4 gene. The Lactobacillus derived from mugwort or Angelica keiskei of this embodiment can promote the expression of the SIRT4 gene.

[0102] The telomerase gene may be at least one selected from the group consisting of TERT gene and TERC gene. The Lactobacillus genus derived from Artemisia argyi or Angelica keiskei of the present embodiment may promote the expression of at least one selected from the group consisting of TERT gene and TERC gene.

[0103] The gene whose expression is regulated by the Lactobacillus from mugwort or Angelica keiskei of this embodiment may be an energy metabolism factor gene. The energy metabolism factor gene may be the PPARGC1A gene. The Lactobacillus from mugwort or Angelica keiskei of this embodiment may promote the expression of the PPARGC1A gene.

[0104] The gene whose expression is regulated by Lactobacillus derived from mugwort or Angelica keiskei of the present embodiment may be at least one selected from a hyaluronic acid biosynthesis factor gene and a hyaluronic acid decomposition factor gene.

[0105] The hyaluronic acid biosynthesis factor gene may be at least one selected from the group consisting of the HAS1 gene, the HAS2 gene, and the HAS3 gene. The Lactobacillus genus from mugwort or Angelica keiskei of this embodiment may promote the expression of at least one selected from the group consisting of the HAS1 gene, the HAS2 gene, and the HAS3 gene.

[0106] The hyaluronan decomposing factor gene may be at least one selected from the group consisting of the HYAL2 gene and the CEMIP gene. The Lactobacillus genus derived from mugwort or Angelica keiskei of the present embodiment may inhibit the expression of at least one selected from the group consisting of the HYAL2 gene and the CEMIP gene.

[0107] The gene regulated by the Lactobacillus from mugwort or tomorrow leaf of the present embodiment can be a basement membrane biosynthesis factor gene. The basement membrane biosynthesis factor gene can be at least one selected from the group consisting of LAMA1 gene, LAMA5 gene, COL4A1 gene and COL7A1 gene. The Lactobacillus from mugwort or tomorrow leaf of the present embodiment can promote the expression of at least one selected from the group consisting of LAMA1 gene, LAMA5 gene, COL4A1 gene and COL7A1 gene.

[0108] The heat shock protein gene expression regulator, medicine or cosmetic of this embodiment promotes the production of type I collagen. The type I collagen production promoter of this embodiment comprises Lactobacillus from mugwort or Angelica keiskei. The type I collagen production promoter of this embodiment comprises Lactobacillus from mugwort or Angelica keiskei and a fermentation liquid from mugwort or Angelica keiskei.

[0109] This embodiment includes Lactobacillus derived from mugwort or Angelica keiskei for promoting the production of type I collagen. This embodiment includes Lactobacillus derived from mugwort or Angelica keiskei for promoting the production of type I collagen and a fermentation liquid derived from mugwort or Angelica keiskei.

[0110] This embodiment includes the use of Lactobacillus derived from mugwort or Angelica keiskei for producing a type I collagen production promoter. This embodiment includes the use of Lactobacillus derived from mugwort or Angelica keiskei and a fermentation liquid derived from mugwort or Angelica keiskei for producing a type I collagen production promoter.

[0111] This embodiment includes a method for promoting the production of type I collagen, comprising administering Lactobacillus from mugwort or Angelica keiskei to a human or non-human animal. This embodiment includes a method for promoting the production of type I collagen, comprising administering Lactobacillus from mugwort or Angelica keiskei and a fermented liquid from mugwort or Angelica keiskei to a human or non-human animal.

[0112] The heat shock protein gene expression regulator, medicine or cosmetic of this embodiment promotes the production of HSP47. The HSP47 production promoter of this embodiment comprises Lactobacillus from Angelica keiskei. The HSP47 production promoter of this embodiment comprises Lactobacillus from Angelica keiskei and a fermentation liquid from Angelica keiskei.

[0113] This embodiment includes Lactobacillus from Angelica keiskei for promoting the production of HSP47. This embodiment includes Lactobacillus from Angelica keiskei for promoting the production of HSP47 and a fermentation liquid from Angelica keiskei.

[0114] This embodiment includes use of Lactobacillus from Angelica keiskei for producing an agent for promoting the production of HSP47. This embodiment includes use of Lactobacillus from Angelica keiskei and a fermented liquid from Angelica keiskei for producing an agent for promoting the production of HSP47.

[0115] This embodiment includes a method for promoting the production of HSP47, comprising administering Lactobacillus from Angelica keiskei to a human or non-human animal. This embodiment includes a method for promoting the production of HSP47, comprising administering Lactobacillus from Angelica keiskei and a fermented liquid from Angelica keiskei to a human or non-human animal.

[0116] The species of the genus Lactobacillus derived from mugwort or ashitaba according to the present embodiment may be at least one species selected from the group consisting of parafarraginis, parabuchneri, buchneri, harbinensis, vini, and nagelii.

[0117] The Lactobacillus genus derived from mugwort or Angelica keiskei of this embodiment may be dead bacteria. The Lactobacillus genus derived from mugwort or Angelica keiskei of this embodiment may be heat-treated. The Lactobacillus genus derived from mugwort or Angelica keiskei of this embodiment may be dried bacterial cells.

[0118] The manufacturing method of the heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment includes obtaining Lactobacillus from mugwort or Angelica keiskei. The manufacturing method of the heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment may further include fermenting mugwort or Angelica keiskei to obtain a fermentation liquid. The manufacturing method of the heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment may further include making the obtained Lactobacillus dead. The manufacturing method of the heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment may further include heating the obtained Lactobacillus. The manufacturing method of the heat shock protein gene expression regulator, medicine or cosmetic of the present embodiment may further include drying the obtained Lactobacillus.

[0119] Example

[0120] Hereinafter, examples of the present invention will be described, but the present invention is of course not limited to the following examples.

[0121] (Example 1: Lactobacillus from Artemisia argyi)

[0122] In mugwort leaves, it is generally believed that the number of lactic acid bacteria reaches the maximum during the period of 2 hours before and after sunrise. In addition, it is generally believed that outside this time period, lactic acid bacteria decrease and photosynthetic bacteria increase. Therefore, during this 2-hour period, a portion of about 20 cm is taken from the front end of the mugwort leaves. The 6.3 kg of mugwort leaves taken are immediately put into the first pickling barrel lined with a plastic bag, and after sprinkling 3.2 kg of molasses and 0.6 kg of coarse salt on the mugwort leaves, the mouth of the plastic bag is closed and sealed. A heavy stone is placed from above the plastic bag to pickle the mugwort leaves.

[0123] After a few days when the pickle juice rises above the mugwort leaves, remove the heavy stone. Next, pour 10L of chlorine-free water for rinsing into the second pickling barrel, and put the pickled mugwort leaves and 10kg of pickle juice into the water. Further prepare a third pickling barrel, and place a metal mesh filter on the opening of the third pickling barrel. Take out the mugwort leaves from the second pickling barrel little by little while rubbing them with your hands, and gently press the mugwort leaves on the metal mesh filter on the opening of the third pickling barrel with your palm to squeeze out the pickle juice.

[0124] After the mugwort leaves are completely squeezed, the pickle remaining in the second pickling barrel is filtered through a metal mesh filter. Next, molasses (Hateruma brown sugar) is dissolved in the pickle in the third pickling barrel to a final concentration of 10% by weight, and crude salt is dissolved to a final concentration of 3% by weight. Thereafter, fermentation is started by setting the ambient temperature of the third pickling barrel to about 30°C. Initially, large bubbles are confirmed to form, which gradually turn into small bubbles, and finally the foaming ends. After about a week, the pH at the end of the foaming is about 3.8. The pickle at this time is used as mugwort fermentation liquid. A portion of the obtained mugwort fermentation liquid is heated at 70°C for 30 minutes to obtain a heat-treated mugwort fermentation liquid after killing the bacteria.

[0125] The unheat-treated Artemisia argyi fermentation broth was analyzed using a next-generation sequencer (MiSeq, Illumina). Figure 1 As shown, the mugwort fermented liquid contains parafarraginis species, parabuchneri species, buchneri species and harbinensis species of the genus Lactobacillus. It should be noted that the next generation sequencer is also called a high throughput sequencer. In addition, Figure 1 The values ​​in the table reflect the bacterial counts of the species contained in the Artemisia argyi fermented liquid.

[0126] (Example 2: Lactobacillus from Angelica keiskei)

[0127] In the Angelica keiskei, it is generally believed that the number of lactic acid bacteria reaches the maximum during the period of 1 hour before and after the sunrise time of the day, which is a total of 2 hours. In addition, it is generally believed that outside this time period, lactic acid bacteria decrease and photosynthetic bacteria increase. Therefore, during this 2-hour period, the stems and leaves of the new buds of the Angelica keiskei are taken. The 6.3 kg of Angelica keiskei taken are immediately put into the first pickling barrel lined with a plastic bag, and after sprinkling 3.2 kg of molasses and 0.6 kg of coarse salt on the Angelica keiskei, the mouth of the plastic bag is closed and sealed. A heavy stone is placed from the top of the plastic bag to pickle the Angelica keiskei.

[0128] After a few days when the pickle juice rises above the ashitaba leaves, remove the heavy stone. Next, pour 10L of chlorine-free water for rinsing into the second pickling barrel, and put the ashitaba leaves pickled and 10kg of pickle juice into the water. Further prepare a third pickling barrel, and place a metal mesh filter on the opening of the third pickling barrel. Take the ashitaba leaves out of the second pickling barrel little by little while washing them by hand, and gently press the ashitaba leaves on the metal mesh filter on the opening of the third pickling barrel with your palm to squeeze out the pickle juice.

[0129] After the Angelica keiskei is completely squeezed, the pickle remaining in the second pickling barrel is filtered through a metal mesh filter. Next, molasses is dissolved in the pickle in the third pickling barrel so that its final concentration is 10% by weight, and crude salt is dissolved so that its final concentration is 3% by weight. Thereafter, fermentation is started by setting the ambient temperature of the third pickling barrel to about 30°C. Initially, large bubbles are confirmed to foam, which gradually turn into small bubbles and finally the foaming ends. After about 1 week, the pH at the end of foaming is about 4.0. The pickle at this time is used as Angelica keiskei fermented liquid. A portion of the obtained Angelica keiskei fermented liquid is heated at 70°C for 30 minutes to obtain a heat-treated Angelica keiskei fermented liquid after killing the bacteria.

[0130] The fermentation broth of Angelica keiskei without heat treatment was analyzed using a next-generation sequencer (MiSeq, Illumina). Figure 2 As shown, the Angelica keiskei fermented liquid contains vini species and nagelii species. Figure 2 The values ​​in the table reflect the bacterial counts of the bacterial species contained in the ashitaba fermented liquid.

[0131] (Example 3: Soymilk fermentation broth using Lactobacillus)

[0132] The soy milk was heated to 70°C and sterilized by heating for about 30 minutes. The unheated mugwort fermented liquid prepared in Example 1 was added to the soy milk after the heat sterilization to a final concentration of about 10 wt%, and the mixture was stirred thoroughly. Thereafter, the soy milk to which the unheated mugwort fermented liquid was added was fermented at 37°C for 24 hours. After fermentation, the solid content was removed by filtration to obtain a soy milk fermented liquid containing the genus Lactobacillus.

[0133] (Example 4: Gene expression examination)

[0134] Live Lactobacillus was isolated from the mugwort fermented liquid obtained in Example 1 and dispersed in pure water at a concentration of 0.05 g / L to prepare Sample 1. In addition, the isolated live Lactobacillus was dispersed in pure water at a concentration of 5.0 g / L to prepare Sample 2. The mugwort fermented liquid containing live Lactobacillus at a concentration of 5.0 g / L was prepared as Sample 3.

[0135] The three-dimensional cultured epidermal model (SkinEtchic RHE: 18RHE 098, EPISKIN) was acclimated overnight using a growth medium (Growth Medium: 18SGM 082, EPISKIN). Next, the three-dimensional cultured epidermal model was transferred to a Transwell chamber (Corning) in a 6-well plate filled with growth medium, and 50 μL of any of samples 1 to 3 was applied to the stratum corneum side of the three-dimensional cultured epidermal model in the well. After 24 hours, the medium with the sample added was removed from the well, and the three-dimensional cultured epidermal model was washed with phosphate buffered saline (PBS(-)) that does not contain calcium and magnesium.

[0136] Use a scalpel to cut the three-dimensional cultured epidermal model together with the membrane from the Transwell chamber, and after immersing the three-dimensional cultured epidermal model in a lysis solution (QIAzol, registered trademark, QIAGEN), a crushing device (Tissue Lyser, QIAGEN) is used to crush the cells to obtain a crushing solution. RNA is purified from the crushing solution using an RNA purification kit (miRNeasy Mini Kit, registered trademark, QIAGEN), and the purified RNA is recovered. The recovered RNA is sent to Mitsubishi Chemical Corporation, which provides commissioned analysis services, and the gene expression in the treated cells in the sample is analyzed using an mRNA expression analysis chip. The gene expression in the untreated cells (control) in the sample is standardized to 1.00, and the ratio of the gene expression in the treated cells in the sample to the gene expression in the control is calculated. In addition, a significant difference test is performed using the Student t test.

[0137] The results are shown in Figure 3 to Figure 8 A value greater than 1.00 indicates that gene expression is promoted compared with the control. A value less than 1.00 indicates that gene expression is inhibited compared with the control.

[0138] (Example 5: Anti-wrinkle effect)

[0139] The mugwort fermented liquid containing the Lactobacillus prepared in Example 1 at a concentration of 5.0 g / L was applied to the cheeks of a 46-year-old woman twice a day for 5 months. Fig. 9 As shown in Example 4, the reduction of wrinkles accompanying aging was confirmed. It should be noted that no inflammatory reaction or tanning was confirmed. As shown in Example 4, the mugwort fermented liquid containing the Lactobacillus prepared in Example 1 at a concentration of 5.0 g / L has a very high effect of promoting the expression of the HSP70 gene, so it is believed that the anti-inflammatory effect and whitening effect of HSP70 exceed the expression promotion effect of inflammation-related genes and tanning-related genes.

[0140] (Example 6: Antibacterial effect of Lactobacillus)

[0141] As Gram-positive cocci, Staphylococcus aureus and MRSA are prepared, as Gram-positive bacilli, Bacillus subtilis and Bacillus cereus are prepared, as Gram-negative cocci, Escherichia coli, Salmonella, Vibrio parahaemolyticus and Klebsiella pneumoniae are prepared, and as Gram-negative bacilli, Pseudomonas aeruginosa is prepared.

[0142] Will be 10 7 0.1 mL of a bacterial solution containing any of the above bacteria at a concentration of 1 / 15 mol / mL was inoculated into 10 mL of a fermented soy milk solution containing Lactobacillus prepared in Example 3, and the solution was allowed to act at 25°C, and the number of viable bacteria of the inoculated bacteria was measured over 24 hours. In addition, as a control, 0.1 mL of the bacterial solution was inoculated into 10 mL of a phosphate buffer solution at a concentration of 1 / 15 mol / L and a pH of 7.2, and the solution was allowed to act at 25°C, and the number of viable bacteria of the inoculated bacteria was measured over 24 hours. The results were as follows: Fig.10 As shown, the soymilk fermented liquid containing Lactobacillus reduced all the prepared bacterial species within 24 hours.

[0143] (Example 7: Antifungal Effect of Lactobacillus)

[0144] As fungi, prepare Trichophyton and Candida. 7 0.1 mL of bacterial solution containing Trichophyton or Candida at a concentration of 1 / 15 mol / mL was inoculated into 10 mL of soy milk fermentation solution containing Lactobacillus prepared in Example 3, and the viable count of the inoculated bacteria was measured over 24 hours at 25°C. In addition, as a control, 0.1 mL of bacterial solution was inoculated into 10 mL of phosphate buffer at a concentration of 1 / 15 mol / L and pH 7.2, and the viable count of the inoculated bacteria was measured over 24 hours at 25°C. The results were as follows. Fig.11 As shown, the soymilk fermented liquid containing Lactobacillus reduced the prepared Trichophyton and Candida within 24 hours.

[0145] (Example 8: Antiviral effect of Lactobacillus)

[0146] As an enveloped virus, a culture solution of influenza A virus (H1N1) was prepared. In addition, as a non-enveloped virus, a culture solution of norovirus (feline calicivirus) was prepared. The virus culture solution was diluted in stages by 10 times each time with purified water. Then, an antiviral test using the soy milk fermentation solution containing Lactobacillus prepared in Example 3 was carried out at room temperature according to 50% tissue culture infection dose (TCID50: 50% Tissue Culture Infectious Dose). The antiviral test was carried out at the Japan Food Analysis Center.

[0147] The results, such as Fig.12As shown in Figure 2, the fermented soybean milk containing Lactobacillus reduced the infection titer of influenza virus within 1 hour. Fig.13 As shown, the soymilk fermentation broth containing Lactobacillus spp. reduced the infection titer of norovirus within 24 hours.

[0148] (Example 9: Anti-Trichophyton Effect of Lactobacillus)

[0149] The unheat-treated mugwort fermented liquid obtained in Example 1 was spray-dried to obtain dry cells of Lactobacillus. The obtained dry cells were suspended in water and glycerol in such a manner that the dry cells were 10 parts by weight to obtain a suspension of Lactobacillus of Example 9. The suspension of Lactobacillus was added to Trichophyton, and the colony forming unit (CFU) of Trichophyton was measured. As a result, Fig.14 As shown, the suspension of Lactobacillus killed Trichophyton within 24 hours.

[0150] (Example 10: Effect of Lactobacillus on Promoting Type I Collagen and HSP47 Production)

[0151] Normal human dermal fibroblasts were maintained in a culture vessel using DMEM medium (+5% FBS) until they reached a confluent state. After reaching confluence, the medium was removed from the culture vessel, and DMEM (0% FBS) containing 600 μmol / L of hydrogen peroxide (H2O2) was added to the culture vessel, and the cells were cultured at 37°C for 1 hour. After culturing for 1 hour, the DMEM (0% FBS) containing H2O2 was removed from the culture vessel, and DMEM medium (+10% FBS) was added to the culture vessel. After repeating this operation for 4 days, the cells were further cultured for 3 days using DMEM (10% FBS), and the resulting cells were used as aging-induced treated cells.

[0152] Senescence-induced cell senescence was confirmed by staining for senescence-associated β-galactosidase (SA-β-Gal), a marker of senescence.

[0153] The senescence-induced cells were cultured at 5.0×10 4The cells were inoculated in a 48-well plate at a cell density of cells / well. After 24 hours of inoculation, the culture medium was replaced with DMEM medium (+0.5% FBS) containing the unheated Artemisia argyi fermented liquid prepared in Example 1 at concentrations of 1.0% and 10.0%, DMEM medium (+0.5% FBS) containing the unheated Angelica keiskei fermented liquid prepared in Example 2 at concentrations of 1.0% and 10.0%, DMEM medium (+0.5% FBS) containing vitamin C magnesium phosphate at 25 μmol / L, DMEM medium (+0.5% FBS) containing vitamin C at 25 μmol / L, and DMEM medium (+0.5% FBS). Then, the cells were cultured for 48 hours and the culture medium was recovered. In addition, after the cells were washed with PBS (-), the cells were trypsinized and the cells were recovered from the culture plate. The recovered cells were disrupted by ultrasound, the obtained cell lysate was centrifuged at 15000 rpm, and the supernatant was recovered.

[0154] The amount of type I collagen in the recovered culture medium was quantified by ELISA (direct method using Anti-Human Collagen Type I antibody (Rabbit)). Fig.15 As shown, when cells were cultured in a medium containing vitamin C magnesium phosphate or vitamin C (positive control), the production of type I collagen was promoted. In addition, when cells were cultured in a medium supplemented with mugwort fermentation liquid and ashitaba fermentation liquid, the production of type I collagen was also promoted.

[0155] In addition, HSP47 in the supernatant of the cell lysate was quantified using a commercially available ELISA kit (abcam). Fig.16 As shown, it was shown that when cells were cultured in a medium supplemented with a fermented broth of Angelica keiskei, the production of HSP47 was promoted.

[0156] (Example 11: Anti-aging effect of Lactobacillus)

[0157] Normal human dermal fibroblasts were cultured at 5.0×10 5 The cells were seeded in 6-well plates at a density of 10 cells / well and cultured for 24 hours.

[0158] The culture medium of each well was removed, and DMEM (0% FBS) containing 600 μmol / L hydrogen peroxide (H2O2) was added to the well, and the cells were cultured at 37°C for 1 hour to induce aging of the cells. After 1 hour of culture, the DMEM (0% FBS) containing H2O2 was removed from the well, and a DMEM medium (+10% FBS) containing 1.0% of the Artemisia argyi fermented liquid of the Lactobacillus genus prepared in Example 1 at a concentration of 5.0 g / L, a DMEM medium (+10% FBS) containing 1.0% of the Angelica keiskei fermented liquid of the Lactobacillus genus prepared in Example 2 at a concentration of 5.0 g / L, a DMEM medium (+10% FBS) containing 10 μmol / L of resveratrol with antioxidant effect as a positive control, and a DMEM medium (+10% FBS) as a negative control were added to the wells. These operations were repeated for 4 days.

[0159] Then, DMEM medium (+10% FBS) containing 1.0% of the Artemisia argyi fermentation broth of Lactobacillus prepared in Example 1 at a concentration of 5.0 g / L, DMEM medium (+10% FBS) containing 1.0% of the Angelica keiskei fermentation broth of Lactobacillus prepared in Example 2 at a concentration of 5.0 g / L, DMEM medium (+10% FBS) containing 10 μmol / L of resveratrol with antioxidant effect as a positive control, and DMEM medium (+10% FBS) as a negative control were added to the wells, and the cells were cultured for 3 days.

[0160] Then, the cells cultured under the respective conditions were cultured at 5.0×10 4 The cells were seeded in a 48-well plate at a density of 10 cells / well and cultured for 24 hours. The cells were then fixed with PBS containing 3% formaldehyde. Next, the solution in the well was replaced with a pH 6 reaction solution containing 1 mg / mL of 5-bromo-4-chloro-3-indolyl-D-galactoside (X-Gal) and allowed to stand for 12 to 16 hours. Then, microscopic observation was performed to observe the staining of senescence-associated β-galactosidase (SA-β-Gal), a marker of aging.

[0161] The results, such as Fig.17 , Fig.18 and Fig.19 As shown, the degree of SA-β-Gal staining was strong in cells induced by aging alone. In contrast, the degree of SA-β-Gal staining was reduced in cells induced by aging and treated with mugwort fermented liquid or ashitaba fermented liquid, similar to cells induced by aging and treated with resveratrol. Therefore, it is shown that mugwort fermented liquid and ashitaba fermented liquid have anti-aging effects.

[0162] (Example 12: Effect of promoting HSP70 production by Lactobacillus)

[0163] The three-dimensional cultured epidermal model (SkinEthic RHE: 19RHE 008, EPISKIN) was acclimated overnight with a proliferation medium (Growth Medium: 19SGM 005, EPISKIN). After acclimation, the epidermal model was transferred to a 6-well plate with 1 mL of the proliferation medium dispensed, and 50 μL of the Artemisia argyi fermented liquid containing the Lactobacillus prepared in Example 1 at a concentration of 5.0 g / L and 50 μL of the Angelica keiskei fermented liquid containing the Lactobacillus prepared in Example 2 at a concentration of 5.0 g / L were applied to the stratum corneum side of the epidermal model.

[0164] After culturing the epidermal model for 24 hours, the fermentation liquid was removed and the epidermal model was washed with PBS (-). The survival rate of the epidermal model was evaluated using the Alamar Blue method. The maintenance culture medium containing 10% Alamar Blue reagent (alamarBlue CellViability Reagent, registered trademark, Invitrogen) was dispensed into a 24-well plate. The epidermal model was transferred to the culture plate and cultured for 2 hours, and then the fluorescence intensity of the culture supernatant was measured. The cell survival rate was calculated as an index (%) relative to the fluorescence intensity of the control group using PBS (-) instead of the fermentation liquid. The results obtained were tested for significant differences using the Student t test. The results, such as Fig. 20 As shown, no significant decrease in cell viability was observed due to the application of the Artemisia argyi fermented liquid or the Angelica keiskei fermented liquid.

[0165] After measuring the fluorescence intensity, the epidermal model was immersed in PBS and the epidermal model was broken using a sample breaking device (Tissue Lyser). The cell lysate was centrifuged at 15000 rpm and the supernatant was recovered. The HSP70 in the recovered solution was quantified using a commercially available ELISA kit (Enzolifesciences). The results were tested for significant differences using the Student t test. The results were as follows: Fig.21 As shown: The production of HSP70 was significantly increased by applying the fermented broth of Artemisia argyi or Angelica keiskei.

[0166] The above-described embodiments and examples are intended to facilitate understanding of the present invention, rather than to restrictively explain the present invention. The present invention can be changed / improved without departing from its purport, and its equivalents are also included in the present invention. That is, as long as the method of appropriately designing and changing the embodiments and examples by those skilled in the art possesses the features of the present invention, it is also included in the scope of the present invention. For example, the elements and the like possessed by the embodiments and examples are not limited to the illustrated contents, and appropriate changes can be made. In addition, the embodiments and examples are illustrative, and of course the configurations shown in different embodiments can be partially replaced or combined, and as long as they include the features of the present invention, they are also included in the scope of the present invention.

Claims

1. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of an ASAH1 gene expression inhibitor.

2. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of a DGAT1 gene expression promoter.

3. Use of a Lactobacillus lactic acid bacterium in the manufacture of a GBA gene expression promoter.

4. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of a CLDN1 gene expression promoter.

5. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of an OCLN gene expression promoter.

6. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of an ITGA2 gene expression promoter.

7. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a CDH1 gene expression promoter.

8. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a CD44 gene expression promoter.

9. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a TLR2 gene expression promoter.

10. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a DEFB1 gene expression promoter.

11. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a DEFB4A gene expression promoter.

12. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a DEFB103A gene expression promoter.

13. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a SIRT4 gene expression promoter.

14. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a TERT gene expression promoter.

15. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a TERC gene expression promoter.

16. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a PPARGC1A gene expression promoter.

17. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a HAS1 gene expression promoter.

18. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a HAS2 gene expression promoter.

19. Use of a lactic acid bacterium of the genus Lactobacillus in the manufacture of a HAS3 gene expression promoter.

20. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of an inhibitor of HYAL2 gene expression.

21. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a CEMIP gene expression inhibitor.

22. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a LAMA1 gene expression promoter.

23. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a LAMA5 gene expression promoter.

24. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a COL4A1 gene expression promoter.

25. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a COL7A1 gene expression promoter.

26. Use of lactic acid bacteria of the genus Lactobacillus in the manufacture of a type I collagen production promoter.

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