KR20240143298A - Composition for enhancing or improving immune activity comprising extracts of artemisia princeps pamp.cv.ssajuari and red ginseng, and health functional food containing the same - Google Patents

Abstract

According to the present invention, disclosed are a composition for enhancing or alleviating immunity and a use thereof, wherein the composition for enhancing or alleviating immunity comprises extracts of indigenous shazuari mugwort and red ginseng of baengnyeongdosan, inhibits the expression of inflammatory cytokines il-1β, il-6, tnf-α, and mcp-1, thereby helping to improve immunity.

Description

{COMPOSITION FOR ENHANCING OR IMPROVING IMMUNE ACTIVITY COMPRISING EXTRACTS OF ARTEMISIA PRINCEPS PAMP.CV.SSAJUARI AND RED GINSENG, AND HEALTH FUNCTIONAL FOOD CONTAINING THE SAME}

The present invention discloses a composition for enhancing or improving immunity, which contains extracts of mugwort and red ginseng as effective ingredients, and a health functional food containing the same.

Immunity refers to the body's defense system that quickly recognizes and eliminates specific pathogens or endotoxins that have invaded the body. A weakened immune system increases the incidence of immune diseases such as asthma, allergies, diabetes, and atopic dermatitis.

Various medicines and foods are being developed and used for the purpose of enhancing immunity. Among them, many studies are being conducted to find various functional raw materials using natural extracts, and various technologies are being proposed for health functional foods using mugwort and red ginseng.

Patent Document 1 discloses a health food composition for enhancing immunity and antioxidation by mixing a red ginseng extract and a fermented herb sugar, and Patent Document 2 proposes a composition for improving allergies by including a composite extract of mugwort, red ginseng, mugwort, fennel, and thuja leaves. In addition, Patent Document 3 mentions a food composition for improving inflammation caused by oral bacterial toxicity by including a fermented Artemisia princeps and a fermented red ginseng concentrated powder.

Looking at these patents, they are compositions containing extracts of mugwort and red ginseng, and each patent uses a different type of mugwort.

Mugwort is a plant belonging to the genus Artemisia in the Asteraceae family. It is a ubiquitous plant that adapts well to climate and soil and is widely distributed throughout the world, with about 200 to 300 species known to exist in the Northern Hemisphere. In general, mugwort is called by its local name for convenience rather than its scientific name, and since plants of the genus Artemisia undergo cross-pollination, even mugworts of the same genus are very different depending on the growth stage and environment. Patent Document 4 discloses a kit for distinguishing mugwort varieties and a method for distinguishing mugwort varieties, and discloses that the genetic characteristics of Sajabal mugwort, Seommae mugwort, and Sajuari mugwort are different through genetic analysis.

Patent documents 5 and 6 disclose compositions for improving skin condition and antibacterial and antifungal compositions containing an extract of Artemisia capillaris treated with a sugar hydrolyzing enzyme as an effective ingredient, and patent documents 7 and 8 mention pharmaceutical compositions for preventing and treating cancer and pharmaceutical compositions for preventing and treating cardiovascular diseases containing an extract of Artemisia capillaris.

KR Publication No. 10-2011-0080470 (Published on 2011.07.13) KR Publication No. 10-2011-0067225 (Published on 2011.06.22) KR Publication No. 10-2016-0073852 (Published on 2016.06.27) KR Publication No. 10-2020-0145895 (Published on December 31, 2020) KR Publication No. 1020170119818 (Published on October 30, 2017) KR Publication No. 1020170119819 (Published on October 30, 2017) KR Publication No. 1020090064708 (Published on 2009.06.22) KR Publication No. 1020090064712 (Published on 2009.06.22)

The present applicant has confirmed that the immune-enhancing effect can be greatly enhanced when using mugwort with red ginseng, focusing on the immune-related effect among the various effects of mugwort. In particular, it has been confirmed that the above effect is further enhanced when mugwort is used alone or mixed with red ginseng, and that the effect can be further enhanced when used as an auxiliary material together with medicinal herbs.

The present invention aims to provide a composition capable of improving immunity, including an extract of mugwort or red ginseng, and a use thereof as a health functional food or pharmaceutical composition.

The present invention provides a composition for enhancing or improving immunity, which contains an extract of mugwort or red ginseng as an effective ingredient, and a health functional food or pharmaceutical composition containing the same.

When a composition according to the present invention is administered to a subject in need of enhancing or improving immunity, the expression or activity of inflammatory cytokines can be reduced.

Cytokines are protein immunoregulators secreted from immune cells and participate in immune responses by binding to specific receptors in the autocrine signaling, paracrine signaling, and endocrine signaling processes. There are various types of cytokines involved in cell proliferation, differentiation, apoptosis, or wound healing, and many of them are particularly involved in immunity and inflammation.

The composition of the present invention uses a natural product that has no human toxicity or side effects, and helps improve infectious diseases and inflammatory diseases, which are immune diseases, by suppressing the expression of inflammatory cytokines (IL-1β, IL-6, TNF-α, and MCP-1), so that it can be used for individuals in need of prevention, treatment, or improvement thereof.

The term "immune disease" as used herein means a disease caused by immunodeficiency, immunosuppression, or damage to the immune system, and may include, but is not limited to, infectious diseases, inflammatory diseases, etc.

The term "subject" as used herein includes a human or any non-human animal. The non-human animal can be, for example, a vertebrate, such as a primate, dog, cow, horse, pig, chicken, duck, goose, rodent, such as a mouse, rat, guinea pig, and the like. The term "subject" is used interchangeably herein with "subject" and "patient".

In this specification, the term “immunity” is used interchangeably with “immune function” and “immune activity”.

The composition for enhancing or improving immunity of the present invention comprises mugwort, or extracts of mugwort and red ginseng, and additionally uses jujube, galgeun, cinnamon, ginger, and licorice as medicinal ingredients for use together.

Sajuari mugwort is a native mugwort from Baeknyeongdo Island. When used together with red ginseng extract, it has a synergistic effect and can suppress the activity of cytokines related to inflammation and allergic reactions, thereby enhancing or improving immunity.

Artemisia princeps Pamp. cv. ssajuari is a type of mugwort. The above-mentioned Artemisia princeps Pamp. cv. ssajuari is rich in calcium and iron, which prevents the body from becoming acidic and enhances resistance to bacteria. It is also known to be good for skin beauty by removing blood stasis and promoting blood circulation, and is effective for patients with neuralgia or muscle pain. The present invention uses an extract of the native Artemisia princeps from Baekryeong Island as the above-mentioned Artemisia princeps. The native Artemisia princeps from Baekryeong Island contains more flavonoid components, which are known to be good for protecting the gastric mucosa and suppressing gastric acid, than common Artemisia, and is rich in cineol and beta-carotine. In particular, the Artemisia princeps from Baekryeong Island contains mucogen-E, which suppresses mediators that cause inflammation and has an antioxidant effect, and this is a component that is differentiated from other common Artemisia princeps.

The mugwort referred to in the present invention includes at least one part of leaves, stems, roots, fruits, and flowers, preferably using leaves, stems, and roots, and more preferably using leaves and stems.

Red ginseng is a type of ginseng that is made by carefully selecting high-quality 6-year-old fresh ginseng grown in suitable ginseng cultivation areas, steaming it for a long time without peeling it, and then drying it. It is light yellow-brown or light reddish-brown in color. The red ginseng helps with recovery from fatigue, boosting immunity, tonic effects, and blood flow by inhibiting platelet aggregation. In addition, as the chemical structure of ginsenoside, which is the main pharmacological action of ginseng, changes during the manufacturing process, new components that were not present in fresh ginseng or were contained in extremely small quantities, such as anticancer, antidiabetic, anti-inflammatory, antioxidant, liver function detoxification, and heavy metal detoxification components, are created or their content increases several times.

Medicinal herbs used along with mugwort and red ginseng include jujube, galgeun, cinnamon, ginger, and licorice.

It is known that jujube strengthens the stomach, helps the meridians and replenishes their deficiencies, soothes the stomach, nourishes the spleen, improves body fluid and energy deficiencies, and harmonizes the properties of all kinds of medicines. In oriental medicine, jujube has the effects of preventing aging, strengthening internal organs, improving neurasthenia, treating anemia, reducing appetite, and making the skin moist, and it is known to have the effect of gently harmonizing the rapid effects of strong medicinal herbs and neutralizing toxicity.

Galgeun is the root of kudzu, used as is or with the pericarp removed. It is mild in nature and sweet in taste. The above galgeun is a root plant that has the efficacy of helping with fever reduction, digestion, hemoptysis, thirst relieving, diarrhea, detoxification, and diuresis in oriental medicine. The above galgeun was originally used in oriental medicine as a diaphoretic and antipyretic. It can be expected to have the effect of relieving high fever and headache, and it is responsible for the antipyretic effect by secreting sweat. In addition, galgeun protects the spleen and stomach, and has the effect of relieving fatigue and replenishing body fluids. Galgeun is rich in vitamins, proteins, carbohydrates, calcium, and other minerals, which strengthen lung function and help relieve headaches and coughs.

Cinnamon is a traditional medicine recorded in the Donguibogam as warming the inside, opening the blood vessels, promoting blood circulation, and strengthening the stomach and intestines. Cinnamon's medicinal ingredients consist of about 1% volatile essential oil components, including phellandrene, eugenol, methyleugual, and cinnamaldehyde. It has been reported to have antibacterial effects, anti-leukoplakia effects, stomachic effects, and saliva and gastric juice secretion stimulation effects, and it is also known to be effective in suppressing cancer and enhancing anticancer immunity.

Ginger is known to have antioxidant effects due to its spicy components such as gingerol, gingerone, and shogaol. Ginger has warm ingredients, so it dissipates cold energy that has invaded the body, warms the digestive system, and stops vomiting and diarrhea caused by the cold energy. In addition, the spicy ingredients and essential oil components in ginger expand blood vessels, purify the blood, and help blood circulation, which is excellent for relieving arthritis, menstrual cramps, and headaches. Ginger prevents cholesterol levels from rising, lowers blood viscosity, and prevents arteriosclerosis. The gingerol component in ginger is known to promote bile and remove cholesterol.

Licorice is a perennial herb belonging to the legume family. Licorice is highly effective in treating general inflammation, gastritis, and stomatitis, and is also used to treat tonsillitis, mastitis, infectious hepatitis, eczema, acne, detoxification, and peptic ulcers. The sweet component of licorice, glycyrrhizin, is a type of saponin glycoside that is broken down to produce glucuronic acid, which binds to toxic substances in the liver and has a detoxifying effect, so it is known to restore liver function and is also capable of treating drug addiction, hepatitis, hives, dermatitis, and eczema.

The composition for enhancing or improving immunity of the present invention comprises extracts of mugwort, red ginseng, jujube, galgeun, cinnamon, ginger, and licorice. The composition may be obtained by mixing and then extracting each herbal medicine, or by obtaining extracts of each herbal medicine and then mixing them. For example, the composition may be obtained by separately obtaining mugwort extract and red ginseng extract and then mixing them, or may be obtained by mixing mugwort and red ginseng and then extracting them.

The term "extract" used herein has the meaning commonly used in the art as a crude extract as described above, but in a broad sense, it also includes a fraction obtained by further fractionating an extract. That is, the extract includes not only one obtained by using the above-described extraction solvent, but also one obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, fractions obtained through various additional purification methods such as separation by various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity) are also included in the extract of the present invention.

As the above extraction solvent, it is preferable to perform hot water extraction using at least one solvent selected from the group consisting of purified water, methanol, ethanol, propanol, butanol, glycerol, propylene glycol, and 1,3-butylene glycol, preferably purified water. The extraction method may use an extraction method such as double boiling extraction, reflux cooling extraction, or ultrasonic extraction. Preferably, the extraction can be performed 1 to 5 times using the hot water extraction method, and the extraction is performed at 80°C to 100°C for 5 to 20 hours, preferably at 100°C for 7 to 12 hours.

The above extraction solvent can be used in an amount of 2 to 50 times the weight of the sample, preferably 2 to 20 times. For extraction, the sample can be left for leaching in the extraction solvent for 1 to 72 hours, specifically 1 to 24 hours. It can also be filtered after extraction.

It may be a result obtained by concentrating the filtered extract under reduced pressure using a vacuum rotary evaporator, but is not limited thereto, and includes all of an extract, a diluted or concentrated extract, a dried product obtained by drying the extract, and a controlled product or purified product thereof.

The extract used in the present invention can be manufactured into a powder state by additional processes such as reduced pressure distillation and freeze drying or spray drying.

The composition for enhancing or improving immunity of the present invention, when mixed with mugwort and red ginseng in a weight ratio of 1:0.2 to 1:1, preferably 1:0.3 to 1:0.7, has a more excellent effect of suppressing the expression or activity of inflammatory cytokines (IL-1β, IL-6, TNF-α, and MCP-1) than when used individually.

Additionally, when medicinal herbs such as jujube, galgeun, cinnamon, ginger, and licorice are used together, the effect of suppressing the expression or activity of inflammatory cytokines (IL-1β, IL-6, TNF-α, and MCP-1) can be further increased.

According to one embodiment, red ginseng is used in an amount of 20 to 100 parts by weight, preferably 30 to 70 parts by weight, per 100 parts by weight of mugwort. In addition, the best effect can be obtained when medicinal herbs are used in an amount of 5 to 30 parts by weight of jujube, 5 to 30 parts by weight of galgeun, 0.5 to 5 parts by weight of cinnamon, 1 to 10 parts by weight of ginger, and 1 to 10 parts by weight of licorice per 100 parts by weight of mugwort.

The composition for enhancing or improving immunity of the present invention may be a food composition. The food may be a health functional food. The term "health functional food" used in this specification means a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and pills using raw materials or ingredients having useful functionality for the human body. Here, "functionality" means obtaining a useful effect for health purposes such as regulating nutrients or physiological actions for the structure and function of the human body.

The above health functional food is not particularly limited as long as it can be consumed to improve antibacterial, anti-inflammatory, anti-viral and immune activity. For example, the above health functional food can be manufactured in the form of a concentrate, powder, granule, pill, tablet, capsule, candy, syrup or beverage, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to a conventional method. The effective ingredient may be used appropriately depending on its intended use (prevention or improvement). In general, when manufacturing food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, relative to the raw material.

However, in the case of long-term intake for health purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range as there is no problem in terms of safety.

There is no particular limitation on the type of the above health functional food. The health functional food composition of the present invention can be manufactured as a food, particularly a functional food. The functional food of the present invention includes ingredients that are usually added during food manufacturing, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the effective ingredient. The natural carbohydrate is preferably a monosaccharide (e.g., glucose, fructose, etc.), a disaccharide (e.g., maltose, sucrose, etc.), an oligosaccharide, a polysaccharide (e.g., dextrin, cyclodextrin, etc.), or a sugar alcohol (e.g., xylitol, sorbitol, erythritol, etc.). The flavoring agent can be a natural flavoring agent (e.g., thaumatin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it may further 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 regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

Examples of foods to which the above health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

The composition for enhancing or improving immunity of the present invention may be a pharmaceutical composition.

The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient and diluent selected from the group consisting of carriers, excipients and diluents. Specifically, the pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods.

In the present invention, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulated, it is prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations are prepared by mixing the extract and its fractions with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives may be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerol gelatin.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" of the present invention means an amount sufficient to treat or prevent a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention. The effective dosage level can be determined according to factors including the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration, the route of administration, and the excretion rate of the composition of the present invention used, the treatment period, drugs combined with or used simultaneously with the composition of the present invention used, and other factors well known in the medical field.

The pharmaceutical composition of the present invention may be administered alone or in combination with a known immunotherapeutic agent. It is important to administer an amount that can achieve the maximum effect with the minimum amount without causing side effects by taking all of the above factors into consideration.

The composition for enhancing or improving immunity according to the present invention can enhance the effect of suppressing the expression or activity of inflammatory cytokines (IL-1β, IL-6, TNF-α, and MCP-1) by containing an extract of mugwort, an extract of red ginseng, and additional herbal extracts. The composition according to the present invention can exhibit a significantly excellent immune-enhancing effect.

In addition, the composition according to the present invention can be used to prevent, treat, or improve immune diseases caused by immunodeficiency, immunosuppression, or immune system damage.

Figure 1 is a graph showing the cytotoxicity of the extract of mugwort + red ginseng and the extract of mugwort + red ginseng + medicinal herb.
Figure 2 is a graph showing the cytotoxicity of mugwort extract, red ginseng extract, and herbal extract.
Figure 3 is a graph comparing the IL-1β expression inhibition efficacy of each extract.
Figure 4 is a graph comparing the IL-6 expression inhibition efficacy of each extract.
Figure 5 is a graph comparing the TNF-α expression inhibition efficacy of each extract.
Figure 6 is a graph comparing the MCP-1 expression inhibition efficacy of each extract.
Figure 7 is a graph comparing the inhibitory efficacy of cytokine inflammatory factors according to the content ratio of mugwort and red ginseng.

Hereinafter, the present invention will be described in more detail by way of examples. However, these examples are intended to exemplify the present invention, and the scope of the present invention is not limited by these examples.

Manufacturing Example 1: Manufacturing of mugwort and red ginseng extracts

100 parts by weight of 5-year-old dried mugwort and 50 parts by weight of 6-year-old red ginseng were washed, dried, cut, and mixed, and then about 10 times the dried weight of the dried mugwort was added and extracted at 100°C for 10 hours to obtain an extract.

Manufacturing Example 2: Manufacturing of mugwort + red ginseng + medicinal herb extracts

An extract was obtained in the same manner as in Manufacturing Example 1 using 50 parts by weight of red ginseng, 9 parts by weight of jujube, 9 parts by weight of galangal, 2 parts by weight of cinnamon, 4 parts by weight of ginger, and 4 parts by weight of licorice per 100 parts by weight of mugwort.

Manufacturing Example 3: Manufacturing of Artemisia princeps extract

An extract was obtained by using only mugwort dried for 5 years in the same manner as in Manufacturing Example 1.

Manufacturing Example 4: Manufacturing of red ginseng extract

An extract was obtained by using only 6-year-old red ginseng and performing the same process as in Manufacturing Example 1.

Manufacturing Example 5: Manufacturing of herbal extracts

An extract was obtained in the same manner as in Manufacturing Example 1 using 9 parts by weight of jujube, 9 parts by weight of galangal, 2 parts by weight of cinnamon, 4 parts by weight of ginger, and 4 parts by weight of licorice.

Manufacturing Example 6: Manufacturing of Artemisia princeps + red ginseng extract

An extract was obtained by performing the same procedure as in Manufacturing Example 1 using Artemisia capillaris instead of Artemisia capillaris.

Test Example 1: Cytotoxicity Evaluation

To evaluate the cytotoxicity (cell viability) of each extract against mouse spleen monocytes, each test substance was diluted and an MTT assay was performed.

[Test method]

After isolating the spleen from C57BL/6J mice (9-10 weeks old), splenocytes were separated using a cell strainer. Ficoll-Plaque ^TM Plus (Cytiva, Cat. 17144002) was placed in a 15 ml tube, and splenocytes were layered on top without mixing with each other, and centrifuged at 400 xg for 30 min. After removing the top layer, the cell layer in the middle of the supernatant was collected, washed with cell culture medium, and the cells were seeded on a cell culture dish. Then, they were stored in an incubator maintained at 37℃, 5% _CO2 , and humidity to allow monocytes to attach to the bottom of the dish. After 4 hours, the cell supernatant was removed, and monocytes were collected by treating with 5 mM EDTA.

Monocytes were seeded in 96-well plates, and each extract was diluted to 1/10, 1/100, 1/1,000 (1k), 1/10,000 (10k), and 1/100,000 (100k) for treatment.

To measure cytotoxicity 24 hours after treatment with the test substance, MTT solution (0.5 mg/ml) was reacted for 4 hours, treated with DMSO for 30 minutes, and then absorbance (OD=570 nm) was measured using an iMarkTM plate reader.

[test]

The results obtained are shown in Table 1, Figures 1 and 2 below. Figure 1 is a graph showing the cytotoxicity of the mugwort extract + red ginseng extract, and the mugwort extract + red ginseng + herbal medicine extract, and Figure 2 is a graph showing the cytotoxicity of the mugwort extract, red ginseng extract, and herbal medicine extract.

　 Cell viability (cell survival rate, %) group 1/10 dilution 1/100 dilution 1/1k dilution 1/10k dilution 1/100k dilution Vehicle 100 100 100 100 100 Ssajjuari mugwort + red ginseng extract 102.11 97.87 98.28 99.17 97.96 Ssajjuari mugwort + red ginseng + medicinal herb extract 105.21 99.79 96.88 99.32 98.54 Ssajjuari mugwort extract 86.08 89.12 92.21 93.21 94.22 Red ginseng extract 85.26 86.74 89.33 90.12 90.15 Herbal extracts 87.56 88.51 88.56 87.22 89.78

According to the above Table 1, Figures 1 and 2, all treatment groups did not show cytotoxicity, and it was confirmed that cell viability actually increased when treated with a high concentration of test substance. This suggests that the extract has a cell proliferation effect.

Test Example 2: Anti-inflammatory effect

In order to evaluate the transcript expression level of inflammatory mediators according to test substance treatment when inducing an inflammatory response in immune cells, the following was performed.

[Test method]

1) Isolation of mouse spleen monocyte:

After isolating the spleen from C57BL/6J mice (9-10 weeks old), splenocytes were separated using a cell strainer. Red blood cells (RBCs) were removed by treating with RBC lysis buffer, and the cells were placed in a cell culture dish and stored in an incubator maintained at 37℃, 5% _CO2 , and humidity to allow monocytes to attach to the bottom of the dish. After 4 hours, the cell supernatant was removed and monocytes were collected by treating with 5 mM EDTA.

2) Drug treatment:

Monocytes were added to a 48-well plate and treated with the diluted extract of the manufacturing example. After 24 hours, LPS (lipopolysaccharide) was treated for 6 hours to induce an inflammatory response.

3) RT-PCR:

Cells induced by inflammatory response were collected and the transcriptome was quantitatively examined by quantitative RT-PCR. RNA was extracted using an RNA isolation kit, and cDNA was synthesized. Transcript expression confirmed four target probe activators in total: IL-1β (Mm00434228_m1), IL-6 (Mm00446190_m1), TNF-α (Mm00443258_m1), and MCP-1 (Mm00441242_m1). The equipment used was ABI 7500 (Applied Biosystems).

The measured activation factors were quantified using the Raji standard curve plot, and the measured value of each activation factor was quantified as a normalized ratio to the quantitative value of GAPDH. The test results of each quantified control group and test substance administration group were calculated as a ratio compared to the quantitative value of the normal group.

[test]

The obtained results are shown in Table 2 below, and Figures 3 to 6 are graphs of the related results. At this time, a 1/10 dilution concentration was used for the test.

　 Relative mRNA (1L-1β/GAPDH) Relative mRNA (1L-6/GAPDH) Relative mRNA (TNF-α/GAPDH) Relative mRNA (MCP-1/GAPDH) Vehicle 50.88 225.55 93.99 9.16 Ssajjuari mugwort + red ginseng extract 0.98 0.01 0.42 1.32 Ssajjuari mugwort + red ginseng + medicinal herb extract 0.82 0.001 0.33 1.01 Ssajjuari mugwort extract 1.22 0.02 1.02 2.45 Red ginseng extract 2.43 0.025 1.82 3.15 Herbal extracts 1.64 0.66 1.52 2.55 Artemisia capillaris + red ginseng extract 2.35 0.54 1.78 2.95

Looking at the results in Table 2 and the figure above, it was confirmed that the extract using mugwort and red ginseng together could increase the effect of inhibiting the expression or activity of inflammatory cytokines (IL-1β, IL-6, TNF-α, and MCP-1).

In addition, if you look at the contents of mugwort and red ginseng extracts, you can see that even though they belong to the same mugwort family, their effects also differ depending on the difference in species.

Test Example 3: Comparison of anti-inflammatory effects according to content ratio

The inhibitory effect test on inflammatory cytokines (IL-6, TNF-α, MCP-1, and IL-1β) was conducted by changing the content ratio of mugwort and red ginseng, and the results are shown in Table 3 and Figure 7 below. The test was performed using the same process as Test Example 2 above.

Weight ratio of mugwort and red ginseng Relative mRNA (1L-1β/GAPDH) Relative mRNA (1L-6/GAPDH) Relative mRNA (TNF-α/GAPDH) Relative mRNA (MCP-1/GAPDH) 1:0.5 0.98 0.01 0.42 1.32 1:0.1 1.15 0.02 0.75 1.87 1:1.5 1.55 0.12 1.75 2.43

Looking at the table and diagram above, it can be seen that when the content ratio of mugwort and red ginseng is 1:0.5, it has a better anti-inflammatory effect than other content ratios.

Claims (6)

A composition for enhancing or improving immunity, comprising an extract of Artemisia princeps Pamp. cv. ssajuari as an effective ingredient.
In the first paragraph,
A composition for enhancing or improving immunity, wherein the extract is extracted with at least one extraction solvent selected from the group consisting of purified water, methanol, ethanol, propanol, butanol, glycerol, propylene glycol, and 1,3-butylene glycol.
In the first paragraph,
Additionally, a composition for enhancing or improving immunity comprising red ginseng extract.
In the first paragraph,
The composition for enhancing or improving immunity is a composition for enhancing or improving immunity that suppresses the expression or activity of inflammatory cytokines IL-1β, IL-6, TNF-α and MCP-1.
A health functional food for enhancing or improving immunity, comprising a composition according to any one of claims 1 to 4.
A pharmaceutical composition for enhancing or improving immunity, comprising a composition according to any one of claims 1 to 4.