KR101595959B1 - Composition for preventing, improving or treating allergic dermatitis comprising hot water extract or ferment extract of chestnut inner shell - Google Patents

Abstract

The present invention provides a pharmaceutical composition for the prevention or treatment of an allergic skin disease by preparing a watery uric extract or a uricellium fermented extract.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing, improving or treating an allergic skin disease, comprising a watery extract of U. P. or a solution of a fermented extract of U. P. [

The present invention relates to a composition for preventing, ameliorating or treating an allergic skin disease containing a watery urine extract or a uricellium fermented extract.

Allergic diseases are increasing in various age groups due to changes in diet and living conditions, exposure to chemical and biological harmful substances due to environmental pollution, and so on. This is a sensitive response to the harmless external environment, and is caused by respiratory diseases such as asthma and rhinitis, or skin diseases such as contact dermatitis and atopic diseases.

In particular, the skin is directly contacted with external allergens by air, food, etc., and causes an immune immune response. Allergic skin diseases are inflammatory diseases that are continuously exposed to external antigens such as food, bacteria, mites, and climate environmental factors. Individual allergic reactions to various environmental factors are different because of their genetic characteristics. In order to improve or treat these diseases, there is a method in which exposure to the allergen causing factors and contact are avoided and the skin is kept clean, and the appropriate medicinal active ingredient is applied to the site of the dermatitis or administered to the whole body.

However, synthetic allergenic dermatitis inhibitors are expensive, and drug and cosmetic methods can not prevent the recurrence of lesions. Therefore, recurrence frequently occurs when external antigens are exposed, and side effects are not proved. The need for the development of therapeutic agents using natural materials is urgent.

Recently, researches and researches on new substances from natural products have been established, and researches on the development of drugs using natural materials have been accelerated. As a result, Mushrooms, and the like, are being actively developed.

Korea has been consuming a lot of fermented foods such as kimchi, soybean paste, soy sauce and so on. Since the physiological activity of fermented foods has recently been known, studies for application as health functional materials have been made. Fermentation can be regarded as a wide range of usable materials production by microorganisms. Microorganisms can take the nutrients from the outside into the cells and maintain and grow life by using the energy generated by decomposition of these nutrients by enzymes. It will. Recently, researchers at the University of California have shown that children who take a supplement containing probiotics can prevent and inhibit allergic skin disease. Among these ingredients, Lactobacillus rhamnosus GG has long-term preventive effects , Respectively.

The term "probiotic" refers to bacteria that are beneficial to the body to help antibiotics in the large intestine. It is a substance that is contained in fermented foods. It is a fermented food that contains various anti-allergic ingredients such as lemon balm, bellflower, ginseng, An attempt has been made to develop a fermented food having improved quality as well as enhanced functionality of the food.

Chestnut (Castanea crenata Sieb) is a deciduous broad-leaved arboreous tree belonging to the oak family. Castane crenata is used for edible, medicinal and processed foods. It is also used as a medicinal material such as raw leaves, chestnut husks, chestnut husks and endothelium. .

Korean Patent Laid-Open No. 10-2010-0088007 discloses a composition for prevention or treatment of fatty liver comprising a uri-fructose ethanol extract. However, the present invention relates to a method for producing a fermented extract and a method for producing an anti-allergic or anti- Have not been taught or implied at all.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for prevention and treatment of allergic skin diseases containing a hydrolyzed extract of U. P. or a U. fermented extract.

The above object of the present invention can be achieved by a method for producing a fermented hot-water extract, comprising the steps of: preparing a hot-water extract of Uru, and fermenting the hot-water extract of Uru- The total polyphenol and total flavonoid content and the antioxidant, anti-inflammatory and anti-atopic effects of the UFI hot water extract and the UFI fermentation extract obtained in the above steps were evaluated through the respective steps.

The present invention has an excellent effect of providing a composition for preventing, ameliorating or treating an allergic skin disease containing a watery uric extract or a uricellium fermented extract.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the results of an experiment in which the total polyphenol content of each of the watery extract of the present invention and the fermented extract of Pui is measured.
FIG. 2 is a graph showing the results of an experiment in which the total flavonoid content of each of the watery fruit extract of the present invention and the fermented extract of Julia pygmae was measured.
FIG. 3 is a graph showing the results of an experiment in which the antioxidative activity was measured by the electron donating ability of each of the watery fruit extract of the present invention and the uri fermented extract.
FIG. 4 is a graph showing the results of an experiment in which antioxidative activity was measured by the superoxide anion radical scavenging ability of each of the watery extract of the present invention and the uri fermented extract.
FIG. 5 is a graph showing the results of an experiment for measuring cytotoxicity of each of the watery fruit extract of the present invention and the fermented extract of Ufu.
FIG. 6 is a graph showing the results of an experiment for measuring the anti-inflammatory activity of each of the watery extract of the present invention and the fermented extract of Julia.
FIG. 7 is a photograph showing the effect of treatment of allergy with treating the atopy-induced mouse of the present invention with the watery extract of the present invention and the yeast extract, respectively.
8 is a graph showing the results of measuring the skin melanin index (A), skin eruption index (B) and skin moisture index (C) from mouse skin tissues after treating each of the hot water or hot water extracts of the present invention with atopy- Fig.
FIG. 9 is a graph showing the results of analysis of the expression of inflammatory genes in mouse skin tissues after treating each of the hot-water extract of U. P. or the U. fermented extract of the present invention with atopy-induced mice.

In the present invention, allergic skin diseases are skin diseases such as contact dermatitis and atopic diseases.

The present invention provides a pharmaceutical composition containing as an active ingredient an udder extract of hot pepper and a fermented extract of Aspergillus sieboldii according to the present invention. The watery extract or watery extract of UFI is preferably contained in an amount of 0.1 to 99% by weight based on the total weight of the pharmaceutical composition.

The pharmaceutical composition according to the present invention may be formulated into various forms including pharmaceutically acceptable carriers, for example oral preparations, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, Can be formulated in the form of sterile injectable solutions.

The pharmaceutically acceptable carrier may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. It also includes diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Solid form preparations for oral use include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose ), Gelatin and the like, and may include a lubricant such as magnesium stearate, talc, and the like. Oral liquid preparations include suspensions, solutions, emulsions, syrups, and the like, and may contain diluents such as water and liquid paraffin, wetting agents, sweetening agents, fragrances, preservatives and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, ethylcellulose, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, And the like, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The present invention provides a health functional food composition comprising as an active ingredient a tubular hot-water extract or a tuberous fermented extract prepared according to the present invention. Preferably, the watery extract or the fermented extract of UFI contains 0.1 to 99% by weight based on the total weight of the health functional food composition.

The food composition of the present invention can be used as a health functional food. The term "functional food" as used herein means a food prepared and processed using a raw material or ingredient having a useful function in the human body pursuant to Law No. 6727 on Health Functional Foods, and the term "functional" And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.

The food composition of the present invention may contain conventional food additives, and the suitability of the above-mentioned "food additives" is not limited by the general rules and general test methods approved by the Food and Drug Administration It shall be determined according to standards and standards for the item.

Examples of the substances found in the above-mentioned "food additives" include natural compounds such as ketones, chemical compounds such as glycine, potassium citrate, nicotinic acid and cinnamic acid, coloring pigments, licorice extracts, crystalline cellulose, high- L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar pigment preparations and the like.

The hard capsule of the capsule type health functional food can be prepared by filling a normal hard capsule with a mixture of herbal medicine extract with an additive such as an excipient or a granular product thereof or a granulated product with a hard gelatinized product. Of the present invention may be filled in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

In case of a ring type health functional food, a mixture of excipient, binder, disintegrant, etc. may be formulated into a herbal medicine extract by an appropriate method. If necessary, the preparation may be mixed with white sugar or other suitable skin care agent or a starch, talc or a suitable substance It is possible to make a ring.

In the granular health functional food, a mixture of an excipient, a binder and a disintegrant may be prepared into a granular form by an appropriate method, and if necessary, a flavoring agent, a mating agent and the like may be contained in the herbal medicine extract. The definitions of the above-mentioned excipients, binders, disintegrants, lubricants, mating agents, flavoring agents and the like of the present invention are described in documents known in the art and include the same or similar functions (see, for example, Korean Association of Pharmacy, 5th ed., Pp. 33-48, 1989).

The present invention provides a cosmetic composition comprising as an active ingredient an extract of U. l. extract prepared according to the present invention or a extract of U. liliaceae. Preferably, the watery extract or watery extract according to the present invention contains 0.1 to 99% by weight based on the total weight of the cosmetic composition.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of spraying, chlorofluorohydrocarbons, propane / Or a propellant such as dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as polyoxyethylene sorbitol ester, microcrystalline cellulose or the like may be used as a carrier component.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate and the like may be used as a carrier component.

The cosmetic composition of the present invention can be applied to face or the like once or several times per day.

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, the embodiments and experimental examples are only for illustrating the present invention, and thus the scope of the present invention is not limited thereto.

< Example  1> invention Julie  Extracts and Production of Fermented Fermented Extracts

The uri used in the present invention was supplied from Foodwell, Daegu, Korea. The fermented milk was fermented with Lactobacillus bifermentans Were used.

Manufacturing example  1: invention Julie  Extract preparation

The extract of Chestnut inner shell (CS) was obtained by adding water 10 times as much as the weight of the sample and repeating the extraction three times at 70 ° C for 3 hours. The above-mentioned hot water extract was concentrated under reduced pressure and then lyophilized and used as a test material for testing the efficacy of the water extract of the present invention.

Manufacturing example  2: invention Julie Fermentation product  Produce

 The L. bifermentans strains used for UFI fermentation were cultured in MRS (Difco, USA) medium and the culture was used as an inoculum.

The fermentation medium for the preparation of the fermented chestnut inner shell (FCS) was prepared by adding 0.5% by weight of the hydrolyzed water extract prepared according to Preparation Example 1 to the whole MRS medium to prepare a mixed medium, The culture broth was inoculated to the whole of the mixed medium at 1% (v / v) and cultured for 24 hours. The cultured medium Centrifuged at 6,000 rpm for 20 minutes, and the supernatant was recovered to obtain a lupus fermentation broth. The above-described fermentation broth was lyophilized to obtain the fermented fermented extract of the present invention and used as a test material for the evaluation of the efficacy of the fermented fermented extract.

< Example  2> invention Julie  Extracts and udder fermented extract of  Efficacy test

Experimental Example  1: Total polyphenol content measurement

Total polyphenol contents of the present invention were measured according to the Folin - Ciocalteu method according to the production methods 1 and 2 of the present invention. Each sample solution was mixed with 0.2 mL of 0.2 N Folin-Ciocalteu reagent diluted 1/10 with 1 mL of each sample solution at 0.25, 0.5, 0.75, and 1 mg / mL, respectively. And reacted at room temperature for 5 minutes. To the reaction solution, 4 mL of 7.5% sodium carbonate (Na ₂ CO ₃ ) solution was added and reacted at room temperature for 1 hour. Absorbance was measured at 765 nm. The content of polyphenols contained in the water extracts of the present invention and the fermented extracts of the present invention was determined from the calibration curves prepared using polyphenol standard substance tannic acid. All experiments were repeated three times.

As shown in FIG. 1, the total polyphenol content was increased by 5 to 10% compared to that of the extract of U. P.

Experimental Example  2: Total flavonoid content measurement

Total flavonoid content was measured by AlCl3 method. In the present invention, 1 mL of the sample solution prepared by mixing the hot water extract (CS) and the yeast extract (FCS) at 0.25, 0.5, 0.75 and 1 mg / mL, respectively, was mixed with 150 μL of a 5% sodium nitrite (NaNO ₂ ) solution After reacting at room temperature for 5 minutes, 300 μL of 10% aluminum chloride (AlCl ₃ ) solution was added, reacted for 5 minutes at room temperature, mixed with 1 mL of 1 M NaOH and absorbance was measured at 510 nm. The calibration curve was prepared using catechin as a reference material. All experiments were repeated three times.

As shown in FIG. 2, there was no significant difference in total flavonoid content between the UFI hot water extract and the UFI fermented extract of the present invention, but it was found that the flavonoid content was increased as the concentration was increased.

Experimental Example  3: Electron donating ability  Measure

The DPPH method was used to measure the antioxidative activity of the urinary insecticidal extract (CS) and the uricellium fermented extract (FCS) sample solution by hydrogen electron donating ability. 12 mg of DPPH was dissolved in 100 mL of ethanol, and the same amount of distilled water was mixed to prepare a DPPH solution. bracket 5 mL of the prepared DPPH reaction solution was added to 0.5 mL of the sample solution at 0.25, 0.5, 0.75, and 1 mg / mL, and the mixture was reacted in a dark place for 15 minutes. Then, the absorbance was measured at 517 nm. The hydrogen electron donating ability was calculated by repeating each experiment three times and calculating the degree of decrease in absorbance of the control group according to the following equation.

Equation 1

EDA (%) = (1-A / B) * 100

A: Absorbance of the sample added group

B: Absorbance of the sample-free group

The DPPH radical scavenging method used for measuring the radical scavenging activity of the UFI hot water extract and the UFI fermented extract of the present invention is a method for measuring the antioxidant ability by reducing the amount of aromatic compounds and aromatic amines .

As shown in FIG. 3, there was no significant difference in the free radical scavenging activity between the hot water extract of the present invention and the hot water extract of the present invention. However, the hot water extract of the present invention showed higher antioxidative activity than the hot water extract of the present invention. Free radical scavenging activity tended to increase with increasing concentration of fermented extracts.

Experimental Example  4: Superoxide anion radical Scatters  Measure

Superoxide anion radical scavenging activity was measured by nitroblue tetrazolium (NBT) reduction method. each Concentrations of 0.25, 0.5, 0.75, and 1 mg / mL To 0.1 mL of the sample solution, add 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.4), 1 mL of the substrate solution containing 0.4 mM xanthine and 0.24 mM NBT, add 1 mL of 0.049 U / mL xanthine oxidase, and incubate at 37 ° C for 20 minutes 1 mL of 1N HCl was added to terminate the reaction, and the absorbance at 560 nm was measured. Superoxide anion radical scavenging activity was calculated by repeating each experiment three times and averaging the superoxide anion radical inhibition rate using the following formula 2 using a control group.

Equation 2

Superoxide anion radical inhibition rate (%) = (1-A / B) * 100

A: Superoxide anion radical production in the sample-added group

B: Superoxide anion radical production in the non-additive group

The NBT method was used to measure the SOD - like ability to convert superoxide radicals into hydrogen peroxide in vivo.

As shown in FIG. 4, it was confirmed that the superoxide anion radical scavenging ability of the present invention was increased as the concentration of both the hot water extract (CS) and the fermented hot water extract (FCS) of the present invention was increased. The fermented extract of the present invention showed a higher Activity. &Lt; / RTI &gt;

Experimental Example  5: Cytotoxicity measurement

HS68 cells were distributed from Korea Cell Line Bank (KCLB) to evaluate the cytotoxicity of UFI hydrolyzate and UFI fermented extract. DMEM (Dulbesco's modified Eagle's media) and FBS (Fetal Bovine Serum) were purchased from WelGENE ), Penicillin-streptomycin was purchased from Gibco (USA), and MTT was purchased from Sigma-Aldrich (USA).

Cell viability was measured by 3- (4,5-dimethylthiazole-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) reduction method. HS68 cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin. Cells were cultured at 37 ° C and 5% CO ₂ . Cells were plated at 3 × 10 ⁴ cells in a 24-well plate and incubated for 24 h at 37 ° C in a 5% CO ₂ incubator. After incubation for 18 hours, 100 μL of the MTT solution dissolved in phosphate buffered saline (PBS) was added to each well. incubator for 4 hours. After culturing, the culture medium was discarded, and 1 mL of DMSO was added thereto. The absorbance was measured at 570 nm.

As shown in FIG. 5, when the extracts of the present invention prepared according to Preparation Example 1 and Preparation Example 2 were treated with 0.1 mg / mL of the hot water extract and the hot water extract of the present invention, the cellulase survival rate And did not show significant cytotoxicity compared with the control group.

Experimental Example  6: Nitric oxide  ( NO ) Production amount measurement

RAW264.7 macrophages were distributed from Korea Cell Line Bank (KCLB) to evaluate the anti-inflammatory effects of the UFI hydrothermal extract and UFI fermented extract. DME (Dulbesco's modified Eagle's media) and FBS (Fetal Bovine Serum) Griess Reagent was purchased from Sigma-Aldrich (USA) and purchased from Korea. Penicillin-streptomycin was purchased from Gibco (USA) and Griess Reagent was purchased from Sigma-Aldrich (USA).

NO concentration was measured by using Griess Reagent System. RAW264.7 macrophages were cultured in a 24-well plate at a concentration of 5 × 10 ⁴ cells / mL and cultured for 24 hours. Various concentrations of uricellium pyroxus extract (CS) and uricellium fermented extract (FCS) Lt; / RTI &gt; After culturing, the cells were treated with 1 μg / mL of LPS and cultured for 18 hours. 100 μL of cell culture solution and 100 μL of Griess reagent were mixed and reacted at room temperature for 15 minutes. Absorbance was measured at 540 nm, and a standard curve was prepared with sodium nitrate to calculate the NO content.

As shown in FIG. 6, the content of NO produced when 1 μg / mL of LPS was treated with RAW 264.7 macrophages tended to decrease in a concentration-dependent manner as the concentration of the hydrolyzate extract and uricellium fermentation extract increased . The NO content of the UFI extract was found to be 16.56 μM at a concentration of 0.1 mg / mL in both samples, which was lower than that of 17.91 μM NO produced in the hot water extract of UFI.

Therefore, it can be concluded that the watery extract and the fermented extract according to the present invention of Preparation Example 1 and Preparation Example 2 have the effect of improving or treating inflammation.

< Example  3> invention Julie  Extracts and udder fermented extract of Anti-atopic  Active black

For the experiment with atopic dermatitis, 1-Chloro-2,4-dinitrobenzene (DNCB), Acetone and Olive oil were used in Sigma-Aldrich (USA). Fibrous Tissue Mini Kit, QuantiTect SYBR Green PCR Kit Was performed by QIAgen Korea (Korea), and cDNA synthesis kit was used by Fermentas (USA).

An experimental animal model

Male NC / Nga mice (21-26 g) at 6 weeks of age were supplied from the Central laboratory animal (Korea) and fed with sufficient feed and water until the day of the experiment. The temperature was 22 ± 2 ℃ and the humidity was 55 ± 15 %, And adapted for one week in an experimental animal kept in 12 hour period contrast and used in the experiment.

After the induction of dermatitis and stabilization of the sample treatment, NC / Nga mice were gently removed from the dorsal area and left to stand for 24 hours to heal the micro wounds of the skin. 200 μL of a 1% DNCB solution (Acetone: Olive oil = 3: 1) was applied to the back region, and 150 μL of 0.4% DNCB solution was applied again after 3 days. After 4 days, 0.4% DNCB solution 150 mu] L for 3 weeks.

Twelve animals were divided into four groups: normal (normal), control (AD), distilled water, 5% UFI hot water extract in the first experimental group (CS) The 5% UFI fermented extracts were applied for 5 days per week for 3 weeks.

Experimental Example  7: Evaluation of skin lesion (visual observation)

The morphological changes such as erythema, dry skin, erythema, edema and hematoma of the skin tissue were visually observed through the application of the urophyte hot water extract and the urophyte fermentation extract after the induction of atopic dermatitis.

The effect of the watery extract of Pucci and the Pucci fermentation extract on the skin lesions of NC / Nga mice was evaluated by applying the Pucci hot water extract and Pucci fermented extract to NC / Nga mice induced by atopic dermatitis through DNCB application for 3 weeks, And the morphological changes of skin lesions were observed.

As a result, the morphological changes of the skin were not observed in the normal group, but the atopic skin changes such as erythema, dry skin, erythema, edema and hematoma were observed in the AD group treated with DNCB. The morphological changes of the skin by DNCB application were partially improved in the CS group treated with the hot water extract of URPI, but there was no significant difference, and the FCS group treated with the UF fermented extract showed a marked improvement and showed a skin shape similar to that of the normal group .

Experimental Example  8: Skin analysis

The skin moisture index, skin melanin index and skin erythema index of atopic dermatitis were measured and compared using MPA5 580. We measured the back skin of NC / Nga mice after 3 consecutive measurements. The measurement was performed at room temperature of 21 ~ 23 ℃ and humidity of 50 ~ 60%.

As a result, the skin melanin index of NC / Nga mice was 44.5 ± 0.17 in normal group, 211.33 ± 12.33 in AD group, 158.67 ± 23.11 in CS group and 99.56 ± 5.2 in FCS group as shown in FIG. 6A And the melanin content increased due to the addition of the fermented extract of the present invention.

The skin eruption index shown in FIG. 6B was 156.5 ± 3.83 in the normal group, 276.33 ± 10.33 in the AD group, 258.44 ± 26.12 in the CS group, and 223.22 ± 4.64 in the FCS group, indicating a decrease tendency of skin erythema . The melanin content and the reduction effect of erythema were confirmed by applying the hot water extract of the present invention, but the decrease effect of the hot water extract of the present invention was more remarkable.

As shown in FIG. 6C, the skin moisture index was 54.6 ± 4.72 in the normal group, 15.5 ± 2.23 in the AD group, 11.5 ± 1.04 in the CS group and 29.2 ± 2.18 in the FCS group, and the skin moisture content decreased by DNCB application Was significantly increased by application of the fermented extract of the present invention.

Therefore, the above experimental results confirm that the present invention can improve the skin lesion change in atopic skin diseases.

Experimental Example  9: Analysis of inflammatory gene expression

Measurement of gene expression of inflammatory cytokines RNA of skin tissue was extracted using Fibrous Tissue Mini Kit. The concentration of extracted RNA was measured using NanoDrop ND-1000. 5 μg of RNA was synthesized by cDNA synthesis kit and then used for real-time PCR. Real-time PCR was performed using Corbett research RG-6000. The expression of the inflammatory cytokine gene was quantitated using the SYBR Green QuantiTect PCR Kit, the endogenous control was performed using GAPDH, and the final concentration of the primer was 10 pM. The nucleotide sequences of the inflammatory cytokine primers used in the real-time PCR are shown in Table 1 below.

Experimental Results The quantitative real-time PCR of IL-1β and TNF-α was performed to confirm whether the anti-atopic effect of the present invention was related to the change of the inflammatory cytokine gene present in the skin. Respectively.

As shown in FIG. 7A, when the RQ value of the normal group was 1, expression of IL-1 beta mRNA was 1.34 in the AD group, IL-1 beta mRNA expression was increased due to atopy, 0.95 in the CS group, 0.82, indicating a tendency to decrease compared to the AD group.

As shown in FIG. 7B, when the RQ value of the normal group was 1, expression of TNF-α mRNA gene was 1.37 in the AD group, 1.1 in the CS group, and 1.06 in the FCS group, And decreased by application of hot - water extract and UFI fermented product.

Thus, it was confirmed that the present invention of the present invention is effective in reducing atopic dermatitis by reducing the expression of inflammatory cytokines such as IL-1β and TNF-α.

As described above, the present invention is an extremely useful invention in the natural medicinal materials industry because it has an excellent effect of providing a composition for the improvement, prevention and treatment of allergic skin disease containing a hot water extract of U.

Claims (5)

Fermented Extract of Lactobacillus acidophilus obtained by Lactobacillus bifermentans Lactobacillus Lactobacillus in a Hot Water Extract of.
A pharmaceutical composition for preventing and treating an allergic skin disease comprising the fermented extract of Ufuk Lactobacillus of claim 1 as an active ingredient.
A functional food composition for preventing and improving atopic dermatitis characterized by containing the fermented extract of Ufuk Lactobacillus of claim 1 as an active ingredient.
A cosmetic composition for improving atopic sensation characterized by containing the fermented extract of Ufuk Lactobacillus of claim 1 as an active ingredient. delete

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