KR20230096823A - Composition for preventing, ameliorating or treating skin disease comprising Cardamine flexuosa extract as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating skin diseases containing an extract of Cardamine flexuosa as an active ingredient, wherein the extract of Stork radish reduces inflammation-inducing chemokine RANTES from keratinocytes, skin barrier and moisturizing factor Since it has the effect of increasing the expression of phosphorus filaggrin and reducing the secretion of histamine released by mast cell degranulation, it can be usefully used as a health functional food, medicine, or feed additive for preventing, improving or treating skin diseases.

Description

Composition for preventing, ameliorating or treating skin disease comprising Cardamine flexuosa extract as effective component}

The present invention relates to a composition for the prevention, improvement or treatment of skin diseases, containing an extract of radish radish as an active ingredient.

The skin of the human body is an indispensable organ for maintaining life by physically and chemically protecting the body from the outside world and at the same time performing biochemical functions necessary for the metabolism of the whole body. All abnormal findings appearing on human skin (including hair, hands, toenails, etc.) are called skin diseases, that is, skin diseases. Since the skin covers the surface of the body, there are many opportunities for direct contact with stimuli from the outside or various pathogens, and is strongly influenced by the body. Moreover, even slight changes in the skin can be seen and touched by hand, and it is easy to take a part of the lesion and histopathologically examine it, or to check for microorganisms, and it is easy to clearly diagnose individual diseases. , there are a considerable number of types compared to other organs, and the names of diseases are complicated and diverse. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, jock itch, psoriasis, herpes simplex/herpes zoster, xeroderma, housewife eczema, and acne. Among them, atopic dermatitis is a recurrent chronic dermatitis with severe itching, the cause of which has not yet been clearly identified, but it is estimated that 10 to 20% of the world's population suffers from this dermatitis.

In addition, the prevalence of atopic dermatitis continues to rise in recent decades due to the increase in airborne allergens caused by environmental pollution and the increase in conditions that form dry skin. Atopic dermatitis is a type of skin inflammatory disease characterized by erythema, edema, severe pruritus and dryness, leaching and scabs, forming blisters in the epidermis in the acute phase and thickening of the skin in the chronic phase. It is a dermatitis with unique clinical and chronic characteristics, and is also called allergic eczema, juvenile febrile fever, and generalized neurodermatitis. In recent decades, the prevalence of atopic diseases has been significantly increasing in developed countries, and in Korea, the number of patients with atopic dermatitis is increasing along with the recent rapid increase in allergic diseases.

On the other hand, Carpesium glossophyllum is distributed throughout the country, and the rhizome leaves remain until flowering. It is pointed, the lower part is narrow, small projections are produced, and there are many hairs on both sides. Stem leaves are alternate, few, rarely hanging, long oval lanceolate smaller than radical leaves, but the upper part is linear lanceolate. It blooms in August-October, and at the end of the stem or branch, one yellowish green head-shaped inflorescence runs downward, and there is a lanceolate glumes. The outer piece is short, turned outward, and has hairs on the outside, and the middle and inner pieces are long oval, with dull ends, and hairy edges. Achenes are 3.5-4mm long, somewhat flat and cone-shaped, and sticky at the end as there is a dot at the end. The rhizome is short and grows sideways.

As a technique related to the extract of stork radish, Korean Patent Publication No. 2021-0145576 discloses a cosmetic composition for moisturizing skin containing an extract of millipede, and Korean Patent No. 1262813 discloses atopic dermatitis containing an extract of Sotae tree as an active ingredient. Alternatively, a composition for preventing and treating allergic skin diseases has been disclosed, but no mention has yet been made of a composition for preventing, improving or treating skin diseases containing the storus extract of the present invention as an active ingredient.

The present invention has been derived from the above needs, and the present invention provides a composition for preventing, improving, or treating skin diseases containing a storian radish extract as an active ingredient, wherein the storian radish extract induces inflammation from keratinocytes. The present invention was completed by confirming that there is an effect of reducing the chemokine RANTES, increasing the expression level of filaggrin, a skin barrier and moisturizing factor, and reducing the secretion of histamine liberated by mast cell degranulation.

In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving skin diseases containing an extract of radish japonica as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases containing the extract of radish radish as an active ingredient.

In addition, the present invention provides a quasi-drug for the prevention or improvement of skin diseases comprising the extract of storus radish as an active ingredient.

In addition, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases containing the extract of radish radish as an active ingredient.

In addition, the present invention provides a cosmetic composition for moisturizing the skin comprising a storus extract as an active ingredient.

In addition, the present invention provides a feed additive for the prevention or improvement of skin diseases containing the storus extract as an active ingredient.

In addition, the present invention provides a veterinary composition for the prevention or treatment of skin diseases containing a storus extract as an active ingredient.

The present invention relates to a composition for the prevention, improvement or treatment of skin diseases containing a storus radish extract as an active ingredient, wherein the stork radish extract reduces the inflammation-inducing chemokine RANTES from skin keratinocytes, improves skin barrier and moisturizing factor, filaggrin. It has the effect of increasing the expression level and reducing the secretion of histamine liberated by mast cell degranulation.

Figure 1 is a result of confirming the cell viability of HaCaT cells in the extract of Dorado radish.
Figure 2 is a result confirming the RANTES production rate of the Stork radish extract in HaCaT cells. ### indicates that the RANTES production rate in the 10 ng/ml TNF-α+10 ng/ml IFN-γ treatment group (TI) increased statistically significantly compared to the control group (Control), p<0.001, *, *** indicates the RANTES production rate in the group treated with 10 ng/ml TNF-α + 10 ng/ml IFN-γ at the same time as the Stork radish extract and 10 ng/ml TNF-α + 10 ng/ml IFN-γ treatment group. Statistically significant decrease, * is p <0.05, *** is p <0.001.
Figure 3 is a result of confirming the expression level of filaggrin in HaCaT cells, Stork radish extract. (A) is the result of treatment of HaCaT cells with radish radish extract, and ** indicates that the filaggrin expression level increased statistically significantly in the control group compared to the control group, or in the positive control group. p<0.01. CF200 is a group treated with 200 μg/ml of radish radish extract, and CaCl ₂ is a positive control group. (B) is 10 ng/ml TNF-α + 10 ng/ml IFN-γ treatment group (TI), and 10 ng/ml TNF-α + 10 ng/ml IFN-γ co-treated group is the result of comparison. ## indicates that there was a statistically significant decrease in the filaggrin expression level in the 10ng/ml TNF-α + 10ng/ml IFN-γ treated group compared to the untreated control group (Control), p<0.01, * ** is 10 ng/ml TNF-α + 10 ng/ml IFN-γ treatment group versus 10 ng/ml TNF-α + 10 ng/ml IFN-γ co-treated group or positive control group. The green expression level was statistically significantly increased, p<0.001.
Figure 4 is the result of confirming the cell viability of MC / 9 cells of the Stork radish extract of the present invention.
Figure 5 is the result of confirming the amount of histamine released according to the treatment of the Stork radish extract in MC / 9 cells. ## indicates a statistically significant increase in the amount of histamine release in the Compound 48/80 treated group compared to the control group, p<0.01, and * indicates a significant increase in the histamine release in the Compound 48/80 treated group versus the Stork radish extract treated group. Histamine release was statistically significantly reduced, p<0.05.
Figure 6 is the result of confirming the amount of TSLP produced according to the treatment of the radish radish extract. ## indicates a statistically significant increase in TSLP production in the 10 ng/ml TNF-α + 10 ng/ml IFN-γ treated group compared to the control group, p<0.01, *, ** indicates 10 ng/ml Compared to the 10ng/ml TNF-α + 10ng/ml IFN-γ treated group, the TSLP production in the 10ng/ml TNF-α + 10ng/ml IFN-γ treated group and the radish extract treated group decreased statistically significantly. , * is p<0.05, ** is p<0.01.

The present invention relates to a health functional food composition for preventing or improving skin diseases, containing an extract of radish radish as an active ingredient.

The skin disease is preferably an allergic or inflammatory skin disease, and the allergic or inflammatory skin disease is skin inflammation, eczema, contact dermatitis, atopic dermatitis, seborrheic dermatitis, lichen simplex chronicus, intertrigo, exfoliative dermatitis, papules It is preferably any one selected from urticaria, psoriasis, psoriatic arthritis, solar dermatitis, sunburn and acne, but is not limited thereto.

It is preferable that the extract of Stork radish is extracted from the outpost of Stork radish, but is not limited thereto, and may be used by adopting a specific part as needed.

The raspberry extract may be prepared by a method comprising the following steps, but is not limited thereto:

(1) extracting by adding an extraction solvent to dried horseradish;

(2) filtering the extract of step (1); and

(3) preparing an extract by drying the filtered extract of step (2).

In the step (1), the extraction solvent is preferably selected from water, C ₁ ~ C ₄ lower alcohol or mixtures thereof, more preferably water or ethanol, but is not limited thereto. In the above production method, as the extraction method, all conventional methods known in the art such as hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. It is preferable to extract the extraction solvent by adding 1 to 20 times the weight of the horseradish. The extraction temperature is preferably 20 to 100 ° C, but is not limited thereto. In addition, the extraction time is preferably 1 to 5 hours, more preferably 2 to 4 hours, but is not limited thereto. In the step (3), drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, more preferably freeze drying, but not limited thereto.

The health functional food composition is preferably prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage, but is not limited thereto. The health functional food composition of the present invention may be prepared by adding the extract of Dorsal japonica as it is or mixing it with other foods or food ingredients, and may be appropriately prepared according to a conventional method. Examples of foods to which the radish radish extract can be added include caramel, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, It may be in the form of any one selected from tea, drinks, alcoholic beverages and vitamin complexes, and includes all health functional foods in the conventional sense. That is, there is no particular limitation on the type of food. The health functional food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, coloring agents and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners , pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. It may also contain fruit flesh for the preparation of natural fruit juices and vegetable beverages. The above components may be used independently or in combination. The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional components. Polysaccharides are sugar alcohols, such as xylitol, sorbitol, and erythritol. The ratio of the natural carbohydrates is not very important, but is preferably 0.01 to 0.04 g, more preferably 0.02 to 0.03 g, with respect to 100 g of the composition of the present invention, but is not limited thereto. As the sweetener, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame may be used.

In addition, the present invention relates to a pharmaceutical composition for the prevention or treatment of skin diseases containing an extract of radish radish as an active ingredient.

The pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent.

The pharmaceutical composition according to the present invention may be a formulation of an ointment, patch or spray applied to the skin, but is not limited thereto.

The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is one commonly used in preparation, and may include saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, water Cross, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate ates, talc, magnesium stearate and mineral oil, and the like, but are not limited thereto. In addition to the above components, antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, or preservatives may be further included. A suitable dosage of the pharmaceutical composition of the present invention will be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, morbid condition, food, administration time, administration route, excretion rate and reaction sensitivity. can The administration route of the pharmaceutical composition for preventing or treating skin diseases of the present invention may be administered through any generally accepted route as long as it can reach the target tissue. The pharmaceutical composition of the present invention is not particularly limited thereto, but depending on the purpose, intramuscular administration, eye drop administration, intraperitoneal administration, intravenous administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, It may be administered through a route such as intrarectal administration, and may be specifically administered through a route of intramuscular administration.

In addition, the present invention relates to a quasi-drug for the prevention or improvement of skin diseases comprising an extract of radish radish as an active ingredient.

In the present invention, the term 'quasi-drugs' refers to textiles, rubber products or similar products used for the purpose of treating, mitigating, treating or preventing diseases of humans or animals, devices that have weak effects on the human body or do not directly act on the human body, and devices or non-machines and similar items, products falling under one of the agents used for sterilization, insecticidal, and similar purposes to prevent infectious diseases, for the purpose of diagnosing, treating, mitigating, treating, or preventing human or animal diseases Items used for the purpose of pharmacologically affecting the structure and function of humans or animals and items other than tools, machines, or devices other than those used. In addition, the quasi-drugs include external skin preparations and personal hygiene products.

When using the extract of radish radish of the present invention as a quasi-drug additive, the extract may be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use. The skin external preparations are not particularly limited thereto, but may be preferably prepared and used in the form of ointments, lotions, sprays, patches, creams, powders, suspensions, gels or gels. The personal hygiene product is not particularly limited thereto, but may preferably be soap, wet tissue, toilet paper, skin cream, face cream, air freshener gel or cleansing gel. In addition, another example of the quasi-drug of the present invention is a disinfectant cleaner, shower foam, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.

In addition, the present invention relates to a cosmetic composition for the prevention or improvement of skin diseases containing the extract of radish radish as an active ingredient.

The cosmetic composition of the present invention includes ingredients commonly used in cosmetic compositions in addition to the above active ingredients, such as fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents ( foaming agents), fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic actives , A typical adjuvant such as a lipid vesicle, and a carrier. The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, and includes, for example, solutions, suspensions, emulsions, pastes, It can be formulated into gels, creams, lotions, powders, oils, powder foundations, emulsion foundations, wax foundations and sprays. More specifically, the cosmetic composition of the present invention is a skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, eye cream, moisture cream, hand cream, essence, Nutrition Essence, Pack, Cleansing Foam, Cleansing Water, Cleansing Lotion, Cleansing Cream, Body Lotion, Body Cleanser, Soap, Powder, Hair Tonic, Hair Conditioner, Hair Essence, Hair Lotion, Hair Nutrition Lotion, Hair Shampoo, Hair Rinse, Hair Treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair drying agent, hair preservative agent, hair dye, hair waving agent, Hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse or hair spray may be prepared as a cosmetic formulation, but is not limited thereto. When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as a carrier component this can be used When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additionally chlorofluorohydro propellants such as carbon, propane/butane or dimethyl ether. When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene fatty acid esters of glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan. When the formulation of the cosmetic composition of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester , microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth, and the like can be used.

In addition, the present invention relates to a cosmetic composition for moisturizing the skin comprising an extract of radish radish as an active ingredient.

In addition, the present invention relates to a feed additive for the prevention or improvement of skin diseases containing the extract of radish radish as an active ingredient.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may refer to any natural or artificial diet, one meal, etc., or a component of the one meal meal, suitable for or suitable for consumption by animals. The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more.

In addition, the present invention relates to a veterinary composition for the prevention or treatment of skin diseases, containing an extract of radish radish as an active ingredient.

The veterinary composition of the present invention may further include appropriate excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate , polysorbate 60, methylparaben, propylparaben and mineral oil. The veterinary composition according to the present invention may further include fillers, anti-agglomerating agents, lubricants, wetting agents, spices, emulsifiers, preservatives, and the like. Or it may be formulated using a method well known in the art to provide delayed release, and the dosage form may be a powder, granule, tablet, capsule, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, It may be in the form of a suppository, a sterile injection solution, or a sterile external preparation. An effective amount of the veterinary composition according to the present invention can be appropriately selected according to the individual animal. The severity of the disease or condition, the sensitivity to the active ingredient of the present invention according to the age, weight, health condition or sex of the subject, the route of administration, the period of administration, factors including other compositions that are combined or used simultaneously with the above composition, and other physiological or It can be determined according to factors well known in the veterinary field.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for explaining the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

Example 1. Preparation of Stork radish extract

After refluxing the dried radish root extract twice for 2 hours with 70% (v/v) ethanol, the extract was filtered, and the filtrate was concentrated under reduced pressure and dried to obtain an extract.

Example 2. Confirmation of anti-inflammatory effect in human keratinocytes

(1) Human keratinocyte culture

Human keratinocytes (HaCaT) were cultured in DMEM medium supplemented with heat-inactivated 10% Fetal Bovine Serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin at 37°C, 5 It was cultured in % CO ₂ environment.

(2) Cytotoxicity test

A CCK-8 (Cell Counting Kit-8) assay was performed to confirm the cytotoxicity of the extract of Dorsalt japonica. In a 96-well plate, HaCaT cells were treated with 100 and 200 μg/ml of the extract of Stork radish, and cultured for 24 hours. After 24 hours, CCK-8 was added to the medium at a concentration of 10%, followed by reaction at 37°C for 2 hours, and absorbance at 450 nm using a microplate spectrophotometer (Biorad, Hercules, CA, USA). was measured, and relative cell viability (%) was calculated through comparison with the control group.

As a result, as shown in FIG. 1A, it was found that the radish radish extract had almost no cytotoxicity.

Meanwhile, as shown in FIG. 1B, HaCaT cells were treated with 10 ng/ml of TNF-α + 10 ng/ml of IFN-γ simultaneously with 10, 20, and 50 μg/ml of radish extract, and cultured for 24 hours. As a result of confirming the cell viability, it was found that there was almost no cytotoxicity.

(3) Inhibitory efficacy test of inflammatory chemokine secretion

Human keratinocyte HaCaT (human keratinocyte cell, ATCC, USA) cells were cultured in DMEM (Dulbecco's modified eagle's medium) medium supplemented with FBS (fetal bovine serum) and PS (penicillin-streptomycin), and 5 × 10 ⁴ per well. After culturing the dog cells in a 96-well plate in DMEM medium containing 10% for 18 hours, the medium was removed and replaced with serum-free DMEM, and 10 ng/ml of TNF-α+10 ng/ml of IFN-γ was added. treatment, or 100 and 200 μg/ml of Dorado radish extract; and 10 ng/ml of TNF-α+10 ng/ml of IFN-γ; and cultured for 24 hours. As a positive control, 12 μg/ml of silymarin (SLY) was treated. The RANTES secretion amount was measured according to the manufacturer's method using an ELISA kit (R&D systems, USA) to measure the amount of RANTES secretion present in the cell supernatant. Statistical differences were assessed for significance using Student's t-test.

As a result, as shown in Figure 2, in the group treated with 10 ng / ml of TNF-α + 10 ng / ml of IFN-γ, the RANTES content increased statistically significantly compared to the control group (Control). In contrast, 10, 20, 50, 100, and 200 μg/ml of Stork radish extract; and 10 ng/ml of TNF-α + 10 ng/ml of IFN-γ; in the group simultaneously treated, the RANTES content was significantly decreased statistically.

Example 3. Confirmation of skin barrier improvement and moisturizing effect in human keratinocytes

HaCaT cells were cultured in DMEM (Dulbecco's modified eagle's medium) supplemented with FBS (fetal bovine serum) and PS (penicillin-streptomycin), and 1 × 10 ⁵ cells per well were placed in a 24-well plate in DMEM medium containing 10%. After culturing for 18 hours, the medium was removed, the medium was replaced with serum-free DMEM, and 200 μg/ml of Dorsalt horseradish extract was treated. In addition, 200 μg/ml of Dorado radish extract and 10 ng/ml of TNF-α+10 ng/ml of IFN-γ were simultaneously treated and cultured for 24 hours. The positive control group was treated with 1.2mM CaCl ₂ . Protein was extracted from the cells using a digestion buffer, and the amount of filaggrin expressed was measured using a filaggrin ELISA kit, and the expression level of filaggrin was calculated by quantifying the total protein. Statistical differences were assessed for significance using Student's t-test.

As a result, as shown in FIG. 3A, the expression level of filaggrin in the group treated with the extract of the present invention was statistically significantly increased, and as shown in FIG. 3B, the expression level of filaggrin was 10 ng/ml of TNF-α. +10ng/ml of IFN-γ treatment, and statistically significant increase by simultaneous treatment of 200μg/ml of Stork radish extract and 10ng/ml of TNF-α + 10ng/ml of IFN-γ.

Example 4. Antihistamine effect in MC/9 cells

(1) Cell culture

Mouse mast cell MC/9 cells were purchased from ATCC (American Type Culture Collection, Rockville, MD, USA) and supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) and 100 units/ml of penicillin. , 100 μg/ml of streptomycin was added to DMEM medium at 37° C. and cultured in a 5% CO ₂ environment.

(2) Cytotoxicity test

A CCK-8 (Cell Counting Kit-8) assay was performed to confirm the cytotoxicity of the extract of Dorsalt japonica. In a 96-well plate, MC/9 mast cells were treated with 100 and 200 μg/ml of radish radish extract and cultured. After 24 hours, 10% CCK-8 was added to the medium, followed by reaction at 37° C. for 2 hours, and absorbance was measured at 450 nm using a microplate spectrophotometer (Biorad, Hercules, CA, USA), Relative cell viability (%) was calculated through comparison with the control group.

As a result, as shown in FIG. 4, the storus extract of the present invention showed little cytotoxicity.

(3) Histamine secretion inhibition efficacy test

Cultured MC/9 mouse mast cells were inoculated in a 96-well plate and treated with 25 μg/ml of compound 48/80 (Sigma, USA) and 100 and 200 μg/ml of radish radish extract for 30 minutes. Thereafter, the secretion amount of allergic histamine secreted in the medium was confirmed using an ELISA kit (Oxford Biomedical Research Inc., USA). The 100% production amount was calculated as the difference in histamine secretion between the group not treated with compound 48/80 and the group treated, and the statistical difference was evaluated for significance using Student's t-test.

As a result, as shown in FIG. 5, the allergy-induced group by compound 48/80 showed a statistically significant increase in the amount of histamine secretion, and in contrast, the amount of histamine secretion decreased in the group simultaneously treated with the storus extract of the present invention. .

Example 5. TSLP production analysis

Human keratinocyte HaCaT (human keratinocyte cell, ATCC, USA) cells were cultured in DMEM (Dulbecco's modified eagle's medium) medium supplemented with FBS (fetal bovine serum) and PS (penicillin-streptomycin), and 5 × 10 ⁴ per well. After culturing the dog cells in a 96-well plate in DMEM medium containing 10% for 18 hours, the medium was removed and replaced with serum-free DMEM, and 10 ng/ml of TNF-α+10 ng/ml of IFN-γ was added. process; Alternatively, 10, 20, and 50 μg/ml of Dorado radish extract; and 10 ng/ml of TNF-α+10 ng/ml of IFN-γ; were co-treated and cultured for 24 hours. As a positive control, 12 μg/ml of silymarin (SLY) was treated. The amount of TSLP secretion was measured using an ELISA kit (R&D systems, USA) according to the manufacturer's method, and the amount of TSLP secretion present in the cell supernatant was measured. Statistical differences were assessed for significance using Student's t-test.

As shown in FIG. 6, it was confirmed that the amount of TSLP produced by the treatment with 10 ng/ml of TNF-α + 10 ng/ml of IFN-γ was reduced by the treatment with the Stork radish extract of the present invention.

Claims (13)

A health functional food composition for preventing or improving skin diseases containing an extract of Cardamine flexuosa as an active ingredient. The functional food composition for preventing or improving skin diseases according to claim 1, wherein the skin disease is an allergic or inflammatory skin disease. The method of claim 2, wherein the allergic or inflammatory skin disease is skin inflammation, eczema, contact dermatitis, atopic dermatitis, seborrheic dermatitis, lichen simplex chronicus, intertrigo, exfoliative dermatitis, papular urticaria, psoriasis, psoriatic arthritis, solar dermatitis , Health functional food composition for preventing or improving skin diseases, characterized in that any one selected from sunburn and acne. The health functional food composition for preventing or improving skin diseases according to claim 1, wherein the extract of Stork radish is obtained by extracting the whole plant of Stork radish. The health functional food composition for preventing or improving skin diseases according to claim 1, wherein the extraction solvent of the horseradish extract is water, C ₁ ~ C ₄ lower alcohol, or a mixture thereof. A pharmaceutical composition for the prevention or treatment of skin diseases, containing an extract of radish japonica as an active ingredient. [Claim 7] The pharmaceutical composition for preventing or treating skin diseases according to claim 6, further comprising a pharmaceutically acceptable carrier, excipient, or diluent in addition to the radish radish extract. The pharmaceutical composition for preventing or treating skin diseases according to claim 6, wherein the composition is in the form of an ointment, patch or spray applied to the skin. A quasi-drug for the prevention or improvement of skin diseases, containing an extract of radish japonica as an active ingredient. A cosmetic composition for the prevention or improvement of skin diseases, containing an extract of radish japonica as an active ingredient. A cosmetic composition for moisturizing the skin containing an extract of radish japonica as an active ingredient. A feed additive for prevention or improvement of skin diseases containing sage radish extract as an active ingredient. A veterinary composition for the prevention or treatment of skin diseases, containing an extract of sagebrush as an active ingredient.

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