Composition for enhancing immunity

Abstract

The present invention relates to a composition for enhancing immunity, comprising, as an active ingredient, a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora, and a polysaccharide isolate thereof, or a fraction thereof. According to the present invention, the composition comprising, as an active ingredient, a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora and a polysaccharide isolate thereof, or a fraction thereof, can increase body weight, increase WBC number, restore spleen cell density, enhance spleen cell proliferation ability, and increase immune-related cytokines, especially for innate immunity. The composition can significantly increase the activity of NK cells, and can be used as an immunopotentiator or a food composition for immunopotentiation.

Description

Technical Field

[0001] The present invention relates to a composition for enhancing immunity, which contains a mixture or fraction thereof of a mixed extract containing Angelica sinensis, Ligusticum wallichii and Paeonia lactiflora and a polysaccharide isolate thereof as an active ingredient. Background Art

[0002] Immunity, as an important biological defense system against diseases caused by external pathogens, is considered crucial for the functionality and efficiency of the immune response in eradicating viral infections prevalent due to the 2019 novel coronavirus (COVID-19).

[0003] The immune system can be divided into natural resistance, non-specific immune system, and specific immune system. Natural resistance (the first line of defense) refers to the anatomical and physiological elements that block all invaders, mainly microorganisms, regardless of their type. The non-specific immune system (the second line of defense) is a defense system composed of phagocytic cells that break through natural resistance to eliminate invaders entering the body. The specific immune system (the third line of defense) is an immune system composed of lymphocytes. Among them, the specific immune system is a highly developed immune system with memory and the ability to distinguish self from non-self. In addition, white blood cells form the second or third line of defense, break through the first line of defense, are responsible for dealing with foreign substances entering the body, and regulate the activities of macrophages and lymphocytes during bacterial infections, viral infections, or inflammatory reactions, playing a key role in determining the therapeutic effects of pharmaceuticals. Immunomodulation is divided into immune function enhancement and immune function suppression. Immune function enhancement refers to helping to enhance the reduced human immune function, including the case of enhancing normal immune function. Immune function suppression is the function of suppressing excessive immune responses. It is known that cytokines and nitric oxide, which are immune factors overproduced by macrophages due to the stimulation of external harmful factors, usually such as lipopolysaccharide, can induce chronic inflammation, and as a result, can induce chronic inflammation-related diseases. However, it is known that the increase in immune factors produced by natural products in normal macrophages can enhance immunity and suppress diseases related to immune deficiency.

[0004] Immunity can be roughly divided into two types, namely innate immunity that one has since birth and acquired immunity that is obtained through adaptation to life, etc.

[0005] Innate immunity is also known as natural immunity, which is characterized by reacting non-specifically to antigens and having no specific memory function. The innate immune system includes the skin and mucous tissues that block the invasion of antigens, the often acidic gastric acid, the complement in the blood, etc. As cells, there are macrophages and polymorphonuclear leukocytes responsible for phagocytosis, NK (natural killer) cells that can kill infected cells, etc. In fact, most infections are defended by innate immunity. Acquired immunity is also known as adaptive immunity. It can remember the initially invading antigens, effectively eliminate the antigens through specific reactions when they invade again, and enhance the innate immunity.

[0006] Recently, the field of innate immunity has attracted much attention because the activation of innate immunity can become a fundamental preventive method against various infectious disease pathogens. And if the immunity is high, it is not easy to get sick. Even if one gets sick, one will recover quickly and can maintain a healthy state continuously. Therefore, it is crucial to control immunity in order to maintain health and extend lifespan.

[0007] However, in terms of modern people's living habits, due to stress, lack of exercise, irregular daily routines, unbalanced diet, smoking, etc., the immunity is easily reduced. In terms of the living environment, various public nuisances are spreading, and there are many factors that reduce immunity. Therefore, modern people need to make efforts to enhance their immunity. As described above, research has been conducted on natural substances for enhancing immunity (for example, Patent Document 1), but Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora have not been fully studied, especially the effect of strengthening innate immunity using a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora has not been studied.

[0008] Therefore, the present inventors have confirmed that a mixture or fraction of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate can increase body weight, increase the number of white blood cell counts (WBC), restore spleen cell density, enhance the spleen cell proliferation ability, and immune-related cytokines. Especially for innate immunity, it can significantly increase NK cell activity, thereby confirming that it can be used in a food composition for immune enhancement, and completed the present invention. Summary of the Invention

[0009] Technical Problem to be Solved

[0010] An object of the present invention is to provide a composition for immune enhancement containing a mixture or fraction of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate as an active ingredient.

[0011] Technical Solution

[0012] In order to solve the above problems and achieve the present invention, an embodiment of the present invention is an immunopotentiating composition containing, as an active ingredient, a mixture of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and a polysaccharide isolate thereof or a fraction thereof.

[0013] As a specific example, the immunity is characterized by being innate immunity, acquired immunity, or a combination thereof.

[0014] As another specific example, the extract is characterized by being extracted using a solvent selected from water, a linear or branched alcohol having 1 to 6 carbon atoms, an organic solvent, and a mixed solvent thereof.

[0015] As another specific example, the extract is characterized by containing Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora in a weight ratio of 1:1.2 to 3:1.2 to 3.

[0016] As another specific example, the mixed extract is characterized by being a mixed extract of Angelica sinensis, Ligusticum chuanxiong (Chinese or Japanese variety), and Paeonia lactiflora.

[0017] As another specific example, the polysaccharide isolate is characterized by being an ethanol polysaccharide.

[0018] As another specific example, the ethanol is characterized by being 95% ethanol.

[0019] As another specific example, the fraction is characterized by being a water fraction or an ethanol fraction of the mixture.

[0020] As another specific example, the water fraction or the ethanol fraction is characterized by having T cell proliferation ability.

[0021] As another specific example, the water fraction or the ethanol fraction is characterized by having 130% to 160% T cell proliferation ability at a concentration of 400 μg / ml to 500 μg / ml.

[0022] As another specific example, the mixture is characterized by containing the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate in a ratio of 60 to 70:30 to 40.

[0023] As another specific example, the mixture or its fraction is characterized by containing 0.001% to 90% by weight based on the total weight of the composition.

[0024] As another specific example, the mixture or its fraction is characterized by increasing the activity of NK cells.

[0025] As another specific example, the mixture or its fraction is characterized by increasing body weight, increasing the number of WBCs, restoring splenocyte density, enhancing splenocyte proliferation ability, and increasing immune-related cytokines.

[0026] As another specific example, the cytokine is characterized by being selected from one or more of IL-2, IFN-γ, IL-4, IL-10, TNF-α, IL-12, and IL-6.

[0027] As another specific example, the composition is characterized by being a food composition.

[0028] As another specific example, the food composition is characterized by being prepared into powder, granule, tablet, capsule, syrup, or beverage.

[0029] Another embodiment of the present invention for realizing the present invention is an immunopotentiating use of a composition, the composition comprising a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora and its polysaccharide isolate or its fraction as an active ingredient.

[0030] Another embodiment of the present invention for realizing the present invention is an immunopotentiating method including a step of treating or administering a composition, the composition comprising a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora and its polysaccharide isolate or its fraction as an active ingredient.

[0031] Beneficial effects

[0032] According to the present invention, a composition comprising a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora and its polysaccharide isolate or its fraction as an active ingredient will increase body weight, increase the number of WBCs, restore splenocyte density, enhance splenocyte proliferation ability, and increase immune-related cytokines. Particularly for innate immunity, it can significantly increase NK cell activity and can be used as an immunopotentiator or a food composition for immunopotentiation. Description of the drawings

[0033] Figure 1 The splenocyte proliferation ability in vitro of the mixture according to a preparation example of the present invention was confirmed.

[0034] Figure 2 The cytokine expression level of splenocytes in vitro of the mixture according to a preparation example of the present invention was confirmed.

[0035] Figure 3 The T cell proliferation ability in vitro of the fraction of the mixture according to a preparation example of the present invention was confirmed.

[0036] Figure 4A schematic diagram during the overall experiment of inducing immune decline and administering the mixture (HG) according to an embodiment of the present invention when preparing an animal model according to a preparation example of the present invention is shown.

[0037] Figure 5 A decrease in body weight and induction of immune decline after administering CTX according to a preparation example of the present invention were confirmed, and the in vivo body weight recovery ability when administering the mixture (HG) according to an embodiment of the present invention was confirmed.

[0038] Figure 6 Induction of immune decline after administering CTX according to a preparation example of the present invention was confirmed, and the in vivo WBC recovery ability when administering the mixture (HG) according to a preparation example of the present invention was confirmed.

[0039] Figure 7 An increase in in vivo NK cell activity when administering the mixture (HG) according to a preparation example of the present invention was confirmed.

[0040] In FIG. 8, it was confirmed that the in vivo spleen cell density was restored when administering the mixture (HG) according to a preparation example of the present invention.

[0041] In FIG. 9, it was confirmed that the expression of immune-related cytokines in vivo increased when administering the mixture (HG) according to a preparation example of the present invention.

[0042] Figure 10 The in vivo spleen cell proliferation ability when administering the mixture (HG) according to a preparation example of the present invention was confirmed. Detailed Description

[0043] Each description and embodiment disclosed in the present invention can also be applied to other different descriptions and embodiments. That is, all combinations of various elements disclosed in the present invention belong to the scope of the present invention. In addition, the scope of the present invention is not limited by the following specific description.

[0044] In addition, those skilled in the art can identify or confirm multiple equivalents of the specific embodiments of the present invention described in this application only by using conventional experiments. In addition, these equivalents are intended to be included in the present invention.

[0045] And, throughout the specification of the present invention, when a part "comprises" a structural element, unless there is a particularly contrary record, it means that other structural elements can also be included, rather than excluding other structural elements.

[0046] Hereinafter, the present invention will be described in more detail.

[0047] The present invention is based on the following findings: A mixture or fraction of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate can increase body weight, increase the number of white blood cells (WBCs), restore splenic cell density, enhance the proliferative ability of splenic cells, and increase immune-related cytokines, particularly significantly increase the activity of natural killer (NK) cells related to innate immunity and enhance immunity, and thus can be used as an immunopotentiator and immunostrengthener.

[0048] An embodiment of the present invention for achieving the above object provides an immunopotentiating composition containing, as an active ingredient, a mixture or fraction of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate.

[0049] In the present invention, the term "Angelica sinensis" refers to the dried root of Chinese Angelica (Angelica sinensis (Oliv.) Diels), which is different from the roots of Korean Angelica gigas Nakai of the Apiaceae family, Japanese Angelica acutiloba (Siebold. & Zucc.) Kitag of the Apiaceae family, or Angelica acutiloba (Siebold. & Zucc.) Kitag. var. sugiyamae Hikino of the Apiaceae family in Hokkaido. Chinese Angelica has a warm and moist smell, is sweet and pungent in taste. Generally, compared with Angelica gigas Nakai, Chinese Angelica has a stronger sweet taste and a weaker pungent taste. The efficacy of Chinese Angelica is the hematopoietic effect of generating blood during anemia, and the hematopoietic effect of Angelica made from the root of Chinese Angelica is significant. However, compared with the hematopoietic effect, the Angelica made from the root of Angelica gigas Nakai has a more excellent effect of promoting blood circulation smoothly, and has strong anti-cancer and blood pressure-lowering effects. Pharmacologically, it is known that Angelica sinensis can promote blood flow in the coronary artery and stimulate erythropoiesis.

[0050] In the present invention, the term "Ligusticum chuanxiong" refers to Chinese Ligusticum chuanxiong Hort. or Japanese Cnidium officinale, which is a perennial herb of the Apiaceae family in the dicotyledonous subclass Archichlamydeae and the order Apiales, and is generally cultivated as a medicinal plant. It has effects such as sedation, analgesia, and strengthening, and is used to treat headaches, anemia, gynecological diseases, etc. The rhizomes from September to November are pickled, the leaves and stems are removed, dried in the sun, and then decocted for oral administration, or used as pills or powders.

[0051] In the present invention, the term "Paeonia lactiflora Pallas" refers to a perennial herbaceous plant of the genus Paeonia in the Paeoniaceae family, a dicotyledonous plant that grows in mountainous areas. Its flowers are beautiful and are used in horticulture. In addition, the roots are used as medicinal materials for analgesia, abdominal pain, menstrual pain, amenorrhea, hematemesis, anemia, contusions, etc. In China, it has been cultivated for ornamental purposes since the Qin Dynasty and the Ming Dynasty, and its cultivation history is longer than that of peonies. Dozens of varieties have been recorded from the Song Dynasty to the Qing Dynasty, and it is distributed in Korea, Mongolia, East Siberia, and other places.

[0052] In the present specification, the Paeonia lactiflora Pallas may include one or more selected from Paeonia lactiflora Pallas, Radix Paeoniae Alba, Radix Paeoniae Rubra, Paeonia lactiflora Pall. var. hirta Regel, and Paeonia veitchii Lynch, and can be used without limitation regardless of the processing method.

[0053] The Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora Pallas can be purchased as commercial products, or collected or cultivated from nature and used without limitation.

[0054] In the present invention, the term "extract" is a mixed extract containing the Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora Pallas. The mixed extract of the Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora Pallas can be extracted from various organs of natural, hybrid, and variant plants. Specifically, it can be extracted not only from the roots, aerial parts, stems, leaves, flowers, main bodies of fruits, and skins of fruits, but also from plant tissue cultures.

[0055] The "mixed extract" is a mixed extract that includes all of the Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora Pallas. It can be an extract obtained by first mixing the three substances and then extracting them together, or an extract obtained by separately extracting each substance and then mixing the extracts. In addition, as long as it is a mixed extract manufactured by a method of manufacturing a mixed extract by a known method, it is included without limitation.

[0056] The extract can be used to extract the pulverized matter of each component using a solvent selected from water, straight-chain or branched-chain alcohols having 1 to 6 carbon atoms, organic solvents, and mixed solvents thereof, but is not limited thereto. In addition, the extraction temperature can be in the range of 20°C to 100°C, and the extraction period can be an extract extracted for about 1 hour to 10 days using extraction methods such as hot water extraction, cold maceration extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited to the extraction temperature, extraction time, or extraction method.

[0057] When the solvent of the extract is water, it may include a cold water extract or a hot water extract.

[0058] The organic solvent is not particularly limited and may be a polyol such as glycerol, ethylene glycol, propylene glycol, 1,3-butanediol; a hydrocarbon solvent such as methyl acetate, ethyl acetate, benzene, n-hexane, diethyl ether, dichloromethane, chloroform; and a nonpolar organic solvent such as petroleum ether, methyl acetate, benzene, hexane, chloroform, dichloromethane, dimethyl ether, ethyl acetate.

[0059] The solvent may also include an aqueous solution of an organic solvent, and its concentration is not particularly limited and may be 1 to 99% (v / v), specifically 60 to 98% (v / v), more specifically 80 to 95% (v / v), further specifically 95% (v / v).

[0060] The extract may include an extract solution, a diluted solution or a concentrated solution of the extract solution, a dried product obtained by drying the extract solution, or these crude products or refined products.

[0061] The extract may contain Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora in a weight ratio of 1 to 10:1 to 10:1 to 10, specifically 1 to 5:1 to 5:1 to 5, more specifically 1:1.2 to 3:1.2 to 3, further specifically 1:1.3 to 2:1.2 to 2, further specifically 1:1.4 to 1.7:1.2 to 1.6, further specifically 1:1.5 to 1.7:1.2 to 1.5, and further specifically 1:1.6:1.3.

[0062] As a specific example, the extract may be a mixed extract containing Angelica sinensis, Ligusticum chuanxiong (Chinese chuanxiong or Japanese chuanxiong), and Paeonia lactiflora in a weight ratio of 1:1.6:1.3.

[0063] As a specific example, the extract may be a mixed extract containing Angelica sinensis, Chinese chuanxiong, and Paeonia lactiflora in a weight ratio of 1:1.6:1.3.

[0064] In the extract of the present invention, an extract of a known natural product or a known component for enhancing or strengthening immunity may be selectively added.

[0065] In the present invention, the term "polysaccharide isolate" includes a polysaccharide precipitated by adding a solvent to the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora. Here, the polysaccharide is also called polysaccharide and may include more than one polysaccharide.

[0066] In the present invention, the method for obtaining the polysaccharide isolate is not particularly limited and can be carried out according to methods commonly used in the art. As a non-limiting example of the method, it can be a polysaccharide isolate obtained from the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention by treating the extract with a specific solvent. Specifically, it can be a polysaccharide isolate formed by adding an organic solvent to the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention, but is not limited thereto.

[0067] In the present invention, the type of solvent used to obtain the polysaccharide isolate is not particularly limited, and any solvent known in the art can be used. As a non-limiting example of the solvent, it can be water, a straight-chain or branched-chain alcohol having 1 to 6 carbon atoms, an organic solvent, or a mixed solvent thereof, and these can be used alone or in combination of one or more, but are not limited thereto. In addition, the organic solvent can be one or more selected from dichloromethane, diethyl ether, chloroform, and ethyl acetate, but is not limited thereto.

[0068] Specifically, the polysaccharide isolate can be an ethanol polysaccharide, and more specifically, it can be a 95% ethanol polysaccharide.

[0069] In the present invention, the term "mixture" refers to a mixture obtained by mixing the extracts of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora prepared in the present invention and the polysaccharide isolates obtained by treating the extracts with specific solvents respectively, and the mixing ratio is not particularly limited.

[0070] By using the extracts of each of these plants and their polysaccharide isolates in combination, the composition can achieve significantly superior effects compared to the case of containing only the extract, polysaccharide isolate, or polysaccharide of one plant. In addition, compared with the case of containing plant extracts, their polysaccharide isolates, or their polysaccharides other than Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora, the composition can exhibit significantly superior immune enhancement or strengthening effects.

[0071] When the mixture is a mixture of an extract and a polysaccharide isolate, the weight ratio of the extract to the polysaccharide isolate can be about 40-80:20-60, specifically, it can be 45-75:25-55, more specifically, it can be 50-70:30-50, further specifically, it can be 60-70:30-40, and even more specifically, it can be 60:40, but is not limited thereto.

[0072] In the present invention, the term "its fraction" refers to a product obtained by separating in order to separate a specific component or a specific group of components from a mixture containing various components.

[0073] In the present invention, the separation method for obtaining the fraction is not particularly limited and can be carried out according to methods commonly used in the technical field. As a non-limiting example of the separation method, it can be a fraction obtained by treating a mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention and its polysaccharide with a specified solvent.

[0074] In the present invention, the type of separation solvent for obtaining the fraction is not particularly limited, and any solvent known in the technical field can be used. As non-limiting examples of the separation solvent, it can be water, a straight-chain or branched-chain alcohol having 1 to 6 carbon atoms, an organic solvent, and a mixed solvent thereof, which can be used alone or in combination of one or more, but is not limited thereto. In addition, the organic solvent can be one or more selected from dichloromethane, diethyl ether, chloroform, and ethyl acetate, but is not limited thereto.

[0075] Specifically, it can be a fraction formed by adding water or ethanol to a mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention and its polysaccharide isolate. More specifically, it can be the water fraction or ethanol fraction of a mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate.

[0076] More specifically, it can be the water fraction of a mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate.

[0077] The water fraction or ethanol fraction of the mixture of the present invention is characterized by having the ability to proliferate T cells. Specifically, when the concentration of the water fraction or ethanol fraction of the mixture of the present invention is 400 μg / ml to 500 μg / ml, it can have a T cell proliferation ability of 130% to 160%, and more specifically, can have a T cell proliferation ability of 135% to 155%.

[0078] Based on the total weight of the composition, it can contain 0.0001 to 90% by weight of the mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate or its fraction. Specifically, based on the total weight of the composition, it can contain 30 to 80% by weight of the mixture or its fraction. More specifically, it can contain 35 to 70% by weight, and more specifically, it can contain 40 to 65% by weight, but is not limited thereto.

[0079] The mixture of the extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate or its fraction can increase NK cell activity.

[0080] It is known that the term NK cells act on the spleen and bone marrow and regulate innate immunity in the body, and NK cell activity can be measured by IFN-γ expression.

[0081] Moreover, the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora can increase body weight, increase the number of white blood cells (WBCs), restore splenic cell density, enhance the proliferative ability of splenic cells, and increase immune-related cytokines.

[0082] The immune-related cytokines may be one or more selected from IL-2, IFN-γ, IL-4, IL-10, TNF-α, IL-12, and IL-6.

[0083] In the present invention, the term "improve" means applying the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention to a composition to enhance, strengthen, or fortify immunity.

[0084] For the purpose of enhancing immunity, improving, strengthening, or fortifying immunity, the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora can be added to a food composition, and the food composition may be a health functional food composition.

[0085] The food composition may contain a food pharmaceutically acceptable carrier.

[0086] The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food, and food additives, and the food composition of this type can be prepared into various forms according to conventional methods well known in the art.

[0087] When using the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora as a food additive, the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora can be directly added, or used together with other foods or food ingredients, and can be appropriately used according to common methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health care, or treatment). Generally, when preparing food or beverage, the mixture or fraction of the extracts and polysaccharide isolates of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora is added in an amount of 0.0001 to 90% by weight, preferably 0.001 to 50% by weight in the raw material composition. However, in the case of long-term intake for health care and hygiene purposes or for health regulation purposes, amounts below this range can also be used.

[0088] There is no particular limitation on the type of the food. Examples of the food to which the substance can be added include meat, sausage, bread, chocolate, confectionery, snacks, cookies, pizza, instant noodles, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, etc., and may include all health foods in the general sense.

[0089] The healthy beverage composition of the present invention, like ordinary beverages, may contain various flavoring agents or natural carbohydrates, etc. as additional components. The natural carbohydrates are monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrin and cyclodextrin; sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used. With respect to 100 parts by weight of the composition of the present invention, the proportion of the natural carbohydrates can generally be about 0.001 to 50 parts by weight, specifically, it can be about 0.01 to 30 parts by weight.

[0090] In addition, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerol, alcohols, carbonating agents for carbonated beverages, etc. The proportion of such additives is not important, but is generally selected in the range of 0.01 to 30 parts by weight per 100 parts by weight of the composition of the present invention. In addition, the composition of the present invention may contain the pulp for preparing natural fruit juices, fruit juice beverages, and vegetable beverages. The proportion of such pulp is not important, but is generally selected in the range of 0.01 to 30 parts by weight relative to 100 parts by weight of the composition of the present invention, and these components can be used alone or in combination.

[0091] Examples

[0092] Hereinafter, the present invention will be described in more detail by way of examples. However, these examples are only used to illustrate the present invention exemplarily, and the scope of the present invention is not limited to these examples, which will be obvious to those of ordinary skill in the technical field to which the present invention pertains.

[0093] Example 1. Preparation of the composition

[0094] 1-1. Preparation of a mixture derived from Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora

[0095] 1-1-1. Preparation of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora

[0096] The roots of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora were separately dried in the shade and finely cut and mixed. Then, distilled water 10 times the total weight of the crude drugs (1000 ml of distilled water was added per 100 g of crude drugs) was added, and hot water extraction was carried out at 95 °C for 4 hours to obtain an extract. Then, the extracts were combined, concentrated under reduced pressure, and filtered to obtain a mixed extract.

[0097] At this time, one of Chinese Angelica and Korean Angelica was selected for Angelica sinensis, one of Chinese Ligusticum chuanxiong and Japanese Ligusticum chuanxiong was selected for Ligusticum chuanxiong, and Paeonia lactiflora was from the same source and mixed in various weight ratios to prepare a mixed extract (refer to Table 1).

[0098] 1-1-2. Preparation of polysaccharide isolate of mixed extract

[0099] Polysaccharide isolates were prepared from each of the mixed extracts prepared in Example 1-1.

[0100] Specifically, the mixed extract prepared in Example 1-1 was taken, and 95% ethanol alcohol with a relative volume of 4 times was added thereto. After standing at a temperature below 25 °C for 16 hours, centrifugation was carried out to obtain the precipitated polysaccharide isolate in each mixed extract.

[0101] 1-1-3. Preparation of mixture of mixed extract and polysaccharide isolate

[0102] The mixed extract prepared in Example 1-1 and the polysaccharide isolate prepared in Example 1-2 were mixed to prepare a mixture.

[0103] Specifically, the mixed extract prepared in Example 1-1 and each of the polysaccharide isolates prepared in Example 1-2 were mixed to prepare each mixture. Then, the separately prepared mixtures were filtered with a sterile filter paper (Milli pore membrane, 0.45 μm) to maintain a sterile state.

[0104] At this time, the mixed extract of 1-1 and the polysaccharide isolate of 1-2 were mixed at a weight ratio of 60:40.

[0105] All of them are summarized in Table 1 below.

[0106] [Table 1]

[0107] Experimental group* Angelica sinensis Ligusticum chuanxiong Paeonia lactiflora Ratio Preparation Example 1 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1.6∶1.3 Preparation Example 2 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1.5∶1.2 Preparation Example 3 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶3∶3 Preparation Example 4 Chinese Angelica sinensis Japanese Ligusticum chuanxiong Paeonia lactiflora 1∶1.6∶1.3 Preparation Example 5 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1∶1 Preparation Example 6 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶4∶1 Preparation Example 7 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1∶4 Preparation Example 8 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶3.5∶3.5 Preparation Example 9 Angelica gigas Japanese Ligusticum chuanxiong Paeonia lactiflora 1∶1∶1 Preparation Example 10 Angelica gigas Japanese Ligusticum chuanxiong Paeonia lactiflora 1∶1.6∶1.3 Preparation Example 11 Angelica gigas Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1.6∶1.3

[0108] * Preparation Examples 1 to 11 of each experimental group were prepared by mixing Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora in the corresponding weight ratios for extraction. After preparing the polysaccharide isolates (ethanol polysaccharides) of each extract, although the polysaccharide isolates are not recorded in Table 1 herein, it means the form of the mixture in which the polysaccharide isolates are mixed.

[0109] 1-2. Preparation of fractions using the mixture of the mixed extract and the polysaccharide isolate

[0110] As shown in Table 2 below, fractions of the mixture of the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate were prepared.

[0111] Specifically, for the mixture of the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of Preparation Example 1 and its polysaccharide isolate (ethanol polysaccharide), water or ethanol was used as a solvent to prepare the fraction of the mixture.

[0112] And for the mixed mixture of the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of Preparation Example 5 and its polysaccharide isolate, the fraction of the mixture was prepared using the same solvent as above.

[0113] They were organized as shown in Table 2 below.

[0114] [Table 2]

[0115] Experimental group Angelica sinensis Ligusticum chuanxiong Paeonia lactiflora Ratio Solvent Fraction of Preparation Example 1 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1.6∶1.3 Water / Ethanol Fraction of Preparation Example 5 Chinese Angelica sinensis Chinese Ligusticum chuanxiong Paeonia lactiflora 1∶1∶1 Water / Ethanol

[0116] Example 2. Confirmation of in vitro immune enhancement effect

[0117] 2-1. Confirmation of spleen cell proliferation ability

[0118] The spleen cell proliferation ability of the preparation examples prepared in Example 1 was confirmed.

[0119] Specifically, after collecting mouse spleens to extract spleen cells, a 96-well plate was prepared and inoculated at a cell count of 1×10 6 cells / ml per well. Then, each sample was diluted in a cell culture medium to a final concentration of 500 μg / ml and then cultured at 37°C and 5% CO2 for 36 hours. At the 36th hour, it was treated with WST-8 (cell viability detection kit) and reacted in a light-shielded state at 37°C and 5% CO2 for 1 hour. After the reaction ended, the OD value was measured at an absorbance of 450 nm, and then the measured OD value was calculated. Compared with the control group, the cell proliferation ability values were compared.

[0120] At this time, as a positive control group, 50 ng / ml of PMA and 2 μg / ml of ionomycin were used.

[0121] The results of the confirmed spleen cell proliferation ability of the mixture are shown in Table 3 and Figure 1 as follows.

[0122] [Table 3]

[0123]

[0124] According to Figure 1 and Table 3, it can be confirmed that the mixtures of Preparation Examples 1 to 3 of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and Preparation Example 4 of the mixture of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora of the present invention have excellent spleen cell proliferation ability compared with those mixed in other mixing ratios or Preparation Examples 5 to 11 without using Angelica sinensis.

[0125] Among them, in the case of Preparation Example 1 and Preparation Example 4 in which Angelica sinensis, Ligusticum chuanxiong or Ligusticum wallichii, and Paeonia lactiflora are mixed at a weight ratio of 1:1.6:1.3, the spleen cell proliferation ability increases to about 150%, compared with Preparation Examples 5 to 11 in which it increases by about 100% to 130%, it can be confirmed that there is a significantly excellent spleen cell proliferation ability of at least 20% to at most 50%.

[0126] From this, it can be seen that the mixture prepared from Angelica sinensis, Ligusticum chuanxiong or Ligusticum wallichii, and Paeonia lactiflora has a very excellent effect in terms of immune enhancement.

[0127] 2-2. Confirmation of the expression level of cytokine TNF-α in spleen cells

[0128] Next, the expression levels of cytokines in spleen cells were confirmed for Preparation Examples 1 to 9 prepared in Example 1. Among the cytokines, the expression level was typically confirmed using TNF-α.

[0129] Specifically, mouse spleens were collected to extract spleen cells, and 48-well plates were prepared. After that, a cell suspension was prepared at a concentration of 1×10 6 cells / ml and inoculated into 48 wells. Each sample was diluted to a concentration of 500 μg / ml and cultured under the conditions of 37°C and 5% CO2 for 48 hours. The target antibody to be confirmed was coated on the maximum binding plate of a 96-well plate for ELISA, and then the antibody expression level was measured at an absorbance of 450 nm. The expression levels of cytokines caused by sample treatment were compared, and the results are shown in Table 4 and Figure 2 as follows.

[0130] The positive control group was treated with 50 ng / ml of PMA and 2 μg / ml of ionomycin.

[0131] [Table 4]

[0132]

[0133] The results show that, from Table 4 and Figure 2 it can be confirmed that, compared with Preparation Examples 5 to 9 with different mixing ratios, in the mixture prepared from Chinese Angelica Root, Chinese Chuanxiong Rhizome or Japanese Chuanxiong Rhizome and Paeonia Lactiflora Pall., namely Preparation Examples 1 to 4, the expression value of the cytokine TNF-α in splenocytes increases.

[0134] Therefore, it can be confirmed that the mixture prepared from Chinese Angelica Root, Chinese Chuanxiong Rhizome or Japanese Chuanxiong Rhizome and Paeonia Lactiflora Pall. has very excellent effects in terms of immune enhancement.

[0135] Confirmation of the T cell proliferation efficiency of the 2-3 fraction

[0136] In order to confirm which immunologically active fractions are contained in the mixture prepared in Example 1-1, the immune enhancement effect of the fractions prepared in 1-2 was confirmed by the T cell proliferation ability. For this, a representative comparison was made with the fractions of Preparation Example 1 and Preparation Example 5 as the center.

[0137] Specifically, the Jurkat cell line was cultured in RPMI-1640 medium supplemented with 10% FBS and 1% antibiotic / antifungal agent. A 96-well plate was prepared, and each well was inoculated at a calculated cell number of 1×10 6 cells / ml. After culturing for 36 hours at 37°C and 5% CO2, each sample was diluted and processed to a concentration of 500 μg / ml. At the 47th hour, WST-8 (Cell Viability Detection Kit) was added, and the reaction was carried out for 1 hour at 37°C and 5% CO2 in the dark. After the reaction, the OD value was measured at an absorbance of 450 nm and calculated. Compared with the control group, the cell proliferation ability values were compared and shown in Table 5 and Figure 3 .

[0138] [Table 5]

[0139]

[0140] The results show that, from Table 5 and Figure 3 it can be confirmed that, compared with the fraction of Preparation Example 5 with a different mixing ratio, the fraction of Preparation Example 1, which is a mixture of the mixed extract of Chinese Angelica Root, Chinese Chuanxiong Rhizome and Paeonia Lactiflora Pall. mixed in a weight ratio of 1:1.6:1.3 and its polysaccharide isolate (ethanol polysaccharide), has significantly excellent T cell proliferation ability. In particular, the water fraction or ethanol fraction of the mixture of the present invention has very excellent T cell proliferation abilities of 154% and 135% respectively at a concentration of 400 μg / ml to 500 μg / ml.

[0141] Thus, in the fractions of Preparation Example 1, the water fraction and the ethanol fraction had very excellent T cell proliferation ability, thereby confirming that the water fraction and the ethanol fraction had excellent effects especially in terms of immune enhancement. Furthermore, based on the above results, it was confirmed that the water fraction and the ethanol fraction obtained from the mixture of the present invention exhibited significantly excellent T cell proliferation ability, indicating that the mixture of the present invention, which is the basis of the above fractions, had excellent immune enhancement activity.

[0142] Example 3. Confirmation of in vivo immune enhancement effect

[0143] The mixed extract obtained by mixing Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora at a ratio of 1:1.6:1.3, which was confirmed to have the most excellent effect in Example 2, and the mixture (HG) of Preparation Example 1 in which its ethanol polysaccharide was mixed were selected, and the in vivo immune enhancement effect was confirmed.

[0144] 3-1. Preparation of immune-suppressed animal model

[0145] As experimental animals, 60 10-week-old to 11-week-old Balb / c mice (male, DooyeolBoitech, Korea) were prepared, and the test period was about 3 weeks.

[0146] Specifically, the mice were acclimated for one week while maintaining the temperature at 23±2°C, humidity at 60%±5, and light-dark cycle at 12 hours (light-dark cycle from 8 am to 8 pm), and a maximum of 5 mice were housed in one cage. After one week, one group was assigned to the treatment group based on the body weight of each mouse. The experimental groups are shown in Table 6 below.

[0147] [Table 6]

[0148] Group Number of n CTL (Negative control) 12 CTX (Positive control) 12 HG125 (Mixture HG125mg / kg prepared in Preparation Example 1) 12 HG250 (Mixture of the present invention HG250mg / kg prepared in Preparation Example 1) 12 HG500 (Mixture of the present invention HG500mg / kg prepared in Preparation Example 1) 12

[0149] Next, to prepare an immune-suppressed animal model, 100 mg / kg of CTX (cyclophosphamide) was administered intraperitoneally, and the specific administration method is as Figure 4 shown.

[0150] Specifically, starting 7 days before inducing immune suppression (before CTX administration), HG was pre-administered orally to the experimental groups for 7 days.

[0151] Then, on the 3rd day (from the 8th day to the 10th day), CTX and the mixture HG of Preparation Example 1 were administered simultaneously, and then on the 8th day (from the 11th day to the 18th day), the mixture HG of Preparation Example 1 was administered additionally.

[0152] Then, using the prepared immunocompromised animal model, the weight gain or loss based on the mixture (HG) prepared in Preparation Example 1 of the present invention, blood cell analysis (WBC), NK cell activity (IFN-γ) analysis, spleen H&E staining analysis, and cytokines of spleen cells were confirmed in sequence. The specific results were the same as those in Examples 3-2 to 3-5 below.

[0153] Moreover, the proliferative ability of the spleen cells in Preparation Example 1 was confirmed using a normal animal model, which was the same as in Example 3-6.

[0154] 3-2. Confirming whether immunodeficiency was induced by administering CTX and confirming the weight and WBC recovery according to the mixture (HG) prepared in Preparation Example 1

[0155] First, it was confirmed whether immunodeficiency was normally induced after administering CTX by analyzing body weight and CBC. At this time, the body weight was measured daily.

[0156] The results were as Figure 5 shown. After confirming that CTX 100 mg / kg was administered three times in total, weight loss was observed in all test groups. This indicated that immunodeficiency was induced after administering CTX.

[0157] Then, the mixture (HG) of the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and the polysaccharide isolate (ethanol polysaccharide) prepared in Preparation Example 1 was administered. As a result, it was confirmed that weight recovery was rapid in all test groups compared with the CTX-administered group.

[0158] Next, CBC measurement was performed as follows. Specifically, before sacrificing the mice, all mice were anesthetized with 1-2% isoflurane, and whole blood was collected from the abdominal aorta vein. Then, blood was collected using an EDTA anticoagulant tube (Becton Dickinson, Franklin Lakes, NJ, USA), and the collected blood was analyzed for complete blood count (CBC) using an ADVIA 2120i hematology system (Siemens, Munich, Germany).

[0159] At this time, an increase in WBC indicates immunological function activity.

[0160] The results were as Figure 6 shown. It was confirmed that WBC decreased significantly in the CTX-administered group and immunodeficiency progressed well. Subsequently, it was confirmed that the number of WBC increased significantly in the HG-treated group of Preparation Example 1 of the present invention, and it was confirmed that the immunological ability was restored compared with the CTX-treated group.

[0161] 3-3. Confirming an increase in NK cell activity in the group administered the mixture prepared in Preparation Example 1

[0162] To confirm the effect on innate immunity in immunity by the administration group of the mixture prepared in Preparation Example 1, NK cell activity was confirmed.

[0163] Using the immunodeficient animals prepared in Example 3-1, NK cell activity was confirmed by the following method.

[0164] Before sacrificing the mice, all the mice were anesthetized with 1-2% isoflurane and blood was collected, and then 0.8 mL of blood was transferred into a tube containing heparin (JW Chemicals, Seoul, Korea). After treatment with an activator, the mixture was placed in the tube and gently mixed at room temperature within 1 hour. Then, the supernatant was placed in a 96-well plate and cultured at 37 °C for 24 hours. After the culture, the sample was put into a stained plate, and IFN-γ expression was analyzed using a Murine NK Activity Kit (NK MAX Bio, Seongnam, Korea).

[0165] Among them, NK cells act on the spleen and bone marrow and regulate innate immunity in the body. NK cell activity is measured by IFN-γ expression.

[0166] The results are as Figure 7 shown. It was confirmed that IFN-γ expression decreased rapidly in the CTX administration group, but in contrast, IFN-γ expression in the HG treatment group of Preparation Example 1 of the mixture of the present invention increased significantly by about 3 times or more.

[0167] From this, it can be confirmed that NK cell activity increased significantly in the HG group of the present invention, thereby significantly enhancing innate immunity.

[0168] 3-4. Confirmation of the recovery of splenocyte density in the administration group of the mixture prepared in Preparation Example 1

[0169] It was confirmed whether the splenocyte density recovered in the administration group of the mixture prepared in Preparation Example 1.

[0170] Using the immunodeficient animals prepared in Example 3-1, hematoxylin and eosin staining of the spleen was performed through the following process. Specifically, the spleen tissue isolated from the mice was fixed with 10% neutral buffered formalin (Biosesang, Yongin, Korea), embedded in paraffin, and then cut into 3 μm using a Finesse ME microtome (Thermo Fisher Scientific, Massachusetts, USA). Then, the sections were dried, the paraffin was removed, washed with distilled water, and stained with hematoxylin and eosin solution.

[0171] Then, after scanning the stained sections with a microfocus microscope, white pulse images were compared and analyzed.

[0172] At this time, the cell density of the white pulp is an important item for evaluating immune enhancement, and the results are as Figure 8aand Figure 8b as shown

[0173] As can be seen from the figure, many dead cells were observed in the CTX administration group, and the cell density decreased. In contrast, in the HG administration group of Preparation Example 1 of the mixture of the present invention, an image in which the reduced cell density was restored could be confirmed.

[0174] 3 - 5. Increased cytokine expression was confirmed in the mixture administration group prepared in Preparation Example 1

[0175] The proliferative ability of spleen cells was confirmed in the mixture administration group prepared in Preparation Example 1.

[0176] An immunocompromised animal was confirmed using the animal model prepared in Example 3 - 1, and cytokines in the spleen were analyzed as follows. At this time, IL - 2, IL - 4, IFN - γ, and TNF - α, which are cytokines, were analyzed.

[0177] Specifically, spleen cells isolated from the spleen tissue of experimental mice were sterilized and screened in RPMI1640 medium (Gibco, Grand Island, USA) containing 10% FBS (Gibco, Grand Island, USA), 1% antibiotic and antifungal agent (Gibco, Grand Island, USA). Then, the number of cells per well was calculated as 1×10 6 cells / ml and dispensed into a 48 - well plate so that the total volume was 150 μL. Thereafter, 50 ng / ml of phorbol 12 - myristate 13 - acetate (Merck, Darmstadt, Germany) and 2 μg / ml of ionomycin (Merck, Darmstadt, Germany) diluted in RPMI1640 medium were treated, and cultured at 37 °C for 48 hours. After culturing under 5% CO2 conditions, ELISA analysis (R&D Systems, Minneapolis, USA) was performed using the supernatant to measure the absorbance at 450 nm. At this time, the cytokine expression levels caused by sample treatment were compared, and the results are shown in Figure 9.

[0178] Among them, cytokines are produced by activated lymphocytes, and an increase in cytokines means enhanced immune function.

[0179] As shown in Figure 9, it was confirmed that cytokines in the CTX - induced model increased similarly in the test group and had a correlation.

[0180] Especially in the HG group of Preparation Example 1 of the present invention, the expression of all cytokines, namely IL - 2, IFN - γ, TNF - α, and IL - 4, increased significantly.

[0181] 3 - 6. The proliferative ability of spleen cells was confirmed in the mixture administration group prepared in Preparation Example 1

[0182] The proliferative ability of splenocytes was confirmed in the group administered with the mixture prepared in Preparation Example 1.

[0183] For this experiment, a normal model was used for confirmation instead of the immunodeficient animals prepared in Example 3-1, and the proliferative ability of splenocytes was analyzed as follows. Specifically, after isolating the spleen from the spleen tissue of experimental mice and sterilizing it, it was screened in RPMI 1640 medium (Gibco, Grand Island, USA) containing 10% FBS (Gibco, Grand Island, USA), 1% antibiotic and antifungal agent (Gibco, Grand Island, USA). Then, the number of cells per well was calculated as 1×10 6 cells / ml and dispensed into a 96-well plate so that the total volume was 100 μL. 50 ng / ml phorbol 12-myristate 13-acetate (Merck, Darmstadt, Germany) and 2 μg / ml ionomycin (Merck, Darmstadt, Germany) were diluted in RPMI 1640 medium and incubated at 3

[0184] 7 °C and 5% CO2 for 35 hours, then treated with WST-8 for 1 hour and reacted in the dark. After the reaction, the absorbance OD value at 450 nm was confirmed with a microplate reader (TECAN, Männedorf, Switzerland) and shown in Figure 10 .

[0185] The spleen is an organ for the maturation of immune cells, which means that as the proliferative ability of splenocytes increases, the immune function is improved.

[0186] The results are as shown in Figure 10 It was confirmed that HG of Preparation Example 1 of the mixture of the present invention significantly proliferated splenocytes in all experimental groups.

[0187] In summary, it was confirmed that the mixture of the mixed extract of Angelica sinensis, Ligusticum chuanxiong and Paeonia lactiflora of the present invention and its ethanol polysaccharide as a polysaccharide isolate can enhance immunity. In particular, when the mixed extract of Chinese Angelica, Chinese Ligusticum chuanxiong or Japanese Ligusticum chuanxiong and Paeonia lactiflora mixed in a ratio of 1:1.6:1.3 and its ethanol polysaccharide as a polysaccharide isolate are used in combination, it can more effectively enhance immunity. Among them, the effect of significantly increasing NK cell activity and enhancing innate immunity is very excellent. Furthermore, the water component and ethanol component of the mixture are very excellent in enhancing immunity.

[0188] 3-7. Statistical analysis

[0189] All of the above data were analyzed using PRISM 7 (Graphpad Software, California, USA). Continuous data were expressed as mean ± S.E.M. Dunnet's multiple comparison test and one-way ANOVA were used to compare consistent variables for all groups. A p-value of < 0.05 was considered statistically significant.

[0190] Thus, the mixture of the mixed extract of Angelica sinensis, Ligusticum chuanxiong and Paeonia lactiflora of the present invention and its ethanol polysaccharide or a fraction of the mixture can be used as an immunopotentiator including innate immunity.

[0191] Based on the above description, those of ordinary skill in the art to which the present invention pertains can understand that the present invention can be implemented in other specific forms without changing the technical idea or essential features of the present invention. In this regard, it should be understood that the embodiments are exemplary in all aspects and not restrictive. The scope of the present invention should be construed such that all changes or variations derived from the meaning and scope of the following claims of the invention and their equivalent concepts are within the scope of the present invention compared to the detailed description.

Claims

1. A composition for enhancing immunity, comprising a mixture of a mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate or a fraction thereof as an active ingredient.

2. The composition according to claim 1, wherein The immunity is innate immunity, acquired immunity, or a combination thereof.

3. The composition according to claim 1, wherein The mixed extract is extracted with a solvent selected from water, a straight-chain or branched-chain alcohol having 1 to 6 carbon atoms, an organic solvent, and a mixed solvent thereof.

4. The composition according to claim 1, wherein, The mixed extract contains Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora in a weight ratio of 1:1.2 - 3:1.2 - 3.

5. The composition according to claim 1, wherein, The mixed extract is a mixed extract of Angelica sinensis, Ligusticum chuanxiong, or Ligusticum wallichii, and Paeonia lactiflora.

6. The composition according to claim 1, wherein The polysaccharide isolate is an ethanol polysaccharide.

7. The composition according to claim 6, wherein, The ethanol is 95% ethanol.

8. The composition according to claim 1, wherein, The fraction is an aqueous fraction or an ethanol fraction of the mixture.

9. The composition according to claim 8, wherein The aqueous fraction or the ethanol fraction has T cell proliferation ability.

10. The composition according to claim 8, wherein, The aqueous fraction or the ethanol fraction has 130% to 160% T cell proliferation ability at a concentration of 400 μg / ml to 500 μg / ml.

11. The composition according to claim 1, wherein, In the mixture, the mixed extract of Angelica sinensis, Ligusticum chuanxiong, and Paeonia lactiflora and its polysaccharide isolate are contained in a ratio of 60 - 70:30 - 40.

12. The composition according to claim 1, wherein, Based on the total weight of the composition, it contains 0.001% by weight to 90% by weight of the mixture.

13. The composition according to claim 1, wherein The mixture or its fraction increases the activity of natural killer cells.

14. The composition according to claim 1, characterized in that, The mixture or its fraction increases body weight, increases the number of white blood cells, restores spleen cell density, enhances spleen cell proliferation ability, and increases immune-related cytokines.

15. The composition according to claim 14, characterized in that, The cytokine is selected from one or more of IL-2, IFN-γ, IL-4, IL-10, TNF-α, IL-12, and IL-6.

16. The composition according to claim 1, wherein, The composition is a food composition.

17. The composition according to claim 16, wherein, The food composition is prepared into powder, granule, tablet, capsule, syrup, or beverage.

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