KR101613693B1 - Composition for Prevention or Treatment of Skin Disease Comprising an Extract of Sargassum Horneri and Method of Preparing The Same - Google Patents

Abstract

The present invention relates to a composition for preventing or treating an atopic skin disease comprising an extract of Horns mellifera as an active ingredient. Since the extract of the present invention can effectively inhibit the production of IgE and the production of IL-4 that induces the production of IgE from B cells, the composition of the present invention comprising the extract of Aspergillus oryzae Can be usefully used for the treatment or prevention of allergic skin diseases and atopic dermatitis.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating an atopic skin disease comprising an extract of Hornsby's mash mash as an active ingredient and a method for preparing the same.

The present invention relates to a composition for preventing or treating an atopic skin disease comprising an extract of Horns mellifera as an active ingredient, and a method for producing the same.

The skin of the human body is physically and chemically an indispensable body for maintaining the life from the outside world while maintaining the biochemical functions necessary for the metabolism of the whole body. All abnormalities appearing in human skin are called skin diseases, namely skin diseases. Since the skin covers the surface of the body, there are many opportunities for direct contact with stimuli and various pathogens from the outside world, and it is strongly influenced by the body. Furthermore, slight changes in the skin can be seen by the eyes and can be touched by hand, and it is easy to take a part of the lesion and to examine it histopathologically, to make a test such as searching of microorganisms, and to make diagnosis of individual diseases clear , The number of species compared to other organs is quite large, and the pathology is complex. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, rash, psoriasis, herpes simplex, shingles, dry skin, housekeeping eczema and acne.

The term " atopy " of atopic dermatitis is a term having the meaning of " strange " or " inappropriate " Atopic dermatitis is a relatively common chronic inflammatory skin disease that occurs in infancy and childhood and recurs and resolves. It can be diagnosed by three characteristics of atopic individual or family history, severe itching, and eczema. Infection, mental stress, It can be aggravated by seasonal and climate change, irritation and allergies.

Although the pathogenesis of atopic dermatitis has not yet been clarified, studies on T lymphocytes have shown that high immunogenicity due to the increase of immunoglobulin E (IgE) antibody or decreased cell-mediated immune function Functional deficiency due to fracture differentiation, and blocking of adrenergic receptors present in the skin are presumed to be the cause of the onset.

In order to treat and manage atopic dermatitis, it is common to prescribe a steroid hormone, ie, a topical corticosteroid preparation, at the same time in order to improve the inflammatory reaction, together with a moisturizing agent that maintains moisture on the skin surface in most dermatologic treatments. However, the use of topical corticosteroids for a long period of time causes various skin side effects such as skin atrophy, vasodilation, pigment desalination, and swelling ancestry.

Therefore, many attempts have been made to find therapeutic agents with high therapeutic efficacy and low side effects from natural products proved to be safe. So far, known anti-atopy related substances have been mainly on land plants and herbal medicines such as Danshen, Dahlia, Jasuri, Saururus, Zawungo,

On the other hand, the oceans, which make up about 70% of the earth, have diverse marine life with different metabolism and biological defense system from land animals. Among them, seaweed contains not only a large amount of minerals and vitamins, but also high polysaccharide contents, so it has the effect of adsorbing and discharging heavy metals in the body. It has various phenolic compounds such as antioxidant effect and antimicrobial effect through high phenol content and free radical scavenging ability It is becoming increasingly important as a natural resource having active activity. In particular, brown algae are known to have various physiological activities such as anticancer, anti-blood coagulation and blood cholesterol lowering effect by containing a high concentration of viscous polysaccharides such as fucoidan, alginic acid and laminarin.

Sargassum Horneri ) is a brown algae belonging to the mating family, and lives in the coastal waters of the south of Korea and Japan. It is mainly used as feed. To date, studies have shown that hornblende extracts have the ability to prevent osteoporosis (Yakugaku Zasshi 2006, 126, 1117-1137) and have been shown to prevent blood clotting (Bioresour. Technol., 2007, 98, 1711-1716). In addition, it has been reported that the hydrothermal extract of Hornsia melliferae has an inhibitory effect on herpes simplex virus type 1 (Chem. Pharm. Bull., 2001, 49, 484-485., Biol. Pharm. , 730-734). However, it has not yet been reported to be effective in the treatment of skin diseases such as atopic dermatitis.

Therefore, the present inventors have tried to find out the efficacy of the extract of Algerianthus sinensis extract in seaweed for the treatment of atopic dermatitis. As a result, it was found that the extract of horseshoe crab mushroom extract was found to be effective in the experiments using atopic dermatitis-induced mice and in various in vivo experiments using immunoglobulin-E (hereinafter simply referred to as 'IgE') and interleukin- -4 ") secretion inhibitory activity, the present inventors have completed the present invention by confirming that they exhibit very excellent atopic dermatitis treatment or prevention efficacy.

It is an object of the present invention to provide a composition for preventing or treating atopic skin diseases, which comprises a substance capable of preventing or treating atopic skin disease from algae natural biological resources, and a composition for preventing or treating atopic skin disease.

It is another object of the present invention to provide a method for extracting a substance capable of preventing or treating an atopic skin disease from natural biological resources.

The present invention

The present invention provides a composition for preventing or treating an atopic skin disease, which comprises an extract of Horns Bay mash as an active ingredient.

The present invention also

(a) powdering the hoe hoe mackerel;

(b) mixing water with the horseshoe crab powder and stirring at room temperature for 20 to 30 hours; And

(c) centrifuging the extract to obtain an upper liquid.

Since the extract of the present invention can effectively inhibit the production of IgE and the production of IL-4 that induces the production of IgE from B cells, the composition of the present invention comprising the extract of Aspergillus oryzae Can be usefully used for the treatment or prevention of allergic skin diseases and atopic dermatitis.

FIG. 1 is a schematic view showing a process for preparing a hornblende mothballs extract by adding ultrapure water to a hornblende powder.
FIG. 2 is a graph showing the proliferation capacity of spleen cells according to the addition of the extract of Horns mellifera to evaluate the cytotoxicity of the extract of Horns sinensis extract of the present invention.
FIG. 3 is a graph showing the amounts of IFN-γ and IL-4 secreted in vitro according to the addition of the concentration of the extract of the present invention.
FIG. 4 is a graph showing the total amount of IgE secreted in serum according to the addition of the extract of Horns mellifera extract according to the present invention in vitro.
FIG. 5 is a photograph showing changes in the external appearance of the skin tissues such as mouse at 1 week and 2 weeks after the application of the extract of the present invention to the atopic dermatitis-like mouse model (negative control: normal mouse, positive control: DNCB (2,4-dinitro chlorobenzene) -containing atopic dermatitis-like mouse, Hornsia mellifera extract-treated group: mice having a mouse model of atopic dermatitis-like mouse model with Hornsia mellifera extract applied thereto)
FIG. 6 is a graph showing the amount of IFN-γ and IL-4 secreted in spleen cells after application of the extract of Horns Cyprinus mellifera of the present invention to atopic dermatitis-like mouse model (negative control, normal mouse, positive control: DNCB (2,4-dinitro chlorobenzene), a group of atopic dermatitis-like mice, and a group of mice treated with a hornblende extract in a mouse model of atopic dermatitis.
FIG. 7 is a graph showing total IgE content in mouse serum after application of the extract of the present invention to the atopic dermatitis-like mouse model (negative control: normal mouse, positive control: DNCB (2,4-dinitro chlorobenzene), a group of atopic dermatitis-like mice, and a group of mice treated with a hornblende extract in a mouse model of atopic dermatitis.
FIG. 8 is a graph showing the proliferation ability of spleen cells after application of the extract of the present invention to the atopic dermatitis-like mouse model (negative control: normal mouse, positive control: DNCB (2,4-dinitro chlorobenzene ), A group of atopic dermatitis-like mice, a group of mice treated with a horn-stem cell extract, in an atopic dermatitis-like mouse model).

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for preventing or treating skin diseases, which comprises an extract of Horns mellifera as an active ingredient.

In the production of the extract of Horns Cygnus sinensis, the dried Horns sinensis can be used. For the efficiency of the extract, the horns can be used in powder form.

Also, an extract obtained by extracting with water as an extracting solvent is preferable for obtaining an extract of Horns mellifera for effective prevention or treatment of skin diseases.

In the following examples of the present invention, a 10-fold amount of ultrapure water was added to powdery hoe saengmyeongbyeol, and the mixture was stirred at room temperature for 24 hours. The mixture was centrifuged to obtain a supernatant, and then the supernatant was concentrated.

Since the extract of Horns Cyprinus mellifera can effectively inhibit the production of IgE and the production of interleukin-4 that induces IgE production from B cells, the composition of the present invention comprising the same as an effective ingredient is useful for preventing or treating skin diseases .

In particular, the skin disease may be a skin disease associated with a high immunoglobulin-E (IgE) antibody level in the serum, preferably, but not exclusively, allergic dermatitis or atopic dermatitis.

The composition of the present invention can be used not only as an effective ingredient, horseshoe extract, but also as a conventionally known preventive or therapeutic agent for skin diseases.

 The composition for preventing or treating skin diseases according to the present invention may be a pharmaceutical composition or a food composition.

The pharmaceutical composition comprising the extract of the present invention as an active ingredient may be prepared using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned effective ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners , A binder, a coating agent, a swelling agent, a lubricant, a lubricant or a flavoring agent.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meats, chocolates, foods, confectionery, pizza, ramen noodles, gums, ice cream, alcoholic beverages, vitamin complexes, .

The food composition of the present invention may contain, as an additional component, various flavors or natural carbohydrates as well as conventional food compositions, in addition to containing the extract of Horns mellifera as an active ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

In addition, the food composition may contain various additives such as flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

The extract of Hornsey mash mash, which is an effective ingredient of the present invention, is a natural substance and has almost no toxicity and side effects. Therefore, it can be safely used for long-term use for skin disease prevention.

The present invention also provides a health functional food for preventing or treating an atopic skin disease comprising an extract of Horns Cygnus sinensis as an active ingredient.

The skin disease of the present invention may be a skin disease associated with a high level of immunoglobulin-E (IgE) antibody in serum, preferably, but not exclusively, allergic dermatitis or atopic dermatitis.

The health functional food for prevention or treatment of skin diseases may be beverage, meat, chocolate, food, confectionery, pizza, ramen, other noodles, gum, ice cream, alcoholic beverage, vitamin complex or health supplement.

The present invention also provides the use of a composition comprising the above-mentioned Horn larvae extract as an active ingredient for the manufacture of a medicament or food for preventing or treating skin diseases. The composition of the present invention comprising the above extract of Horns Cygnus sinensis as an active ingredient can be used for the preparation of medicines or foods for the prevention or treatment of diseases related to the skin.

The present invention also provides a method for the prevention or treatment of an atopic skin disease comprising administering to a mammal a therapeutically effective amount of a hornbill plant extract.

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

As used herein, the term "therapeutically effective amount " refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, This includes an amount that induces relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the treatment method of the present invention, in the case of an adult, it is preferable to administer the extract of the present invention from a dose of 1 mg / kg to 250 mg / kg once or several times a day.

In the treatment method of the present invention, the composition comprising the extract of the present invention as an active ingredient may be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, ≪ / RTI > can be administered in a conventional manner.

The present invention also

(a) powdering the hoe hoe mackerel;

(b) mixing water with the horseshoe crab powder and stirring at room temperature for 20 to 30 hours;

(c) centrifuging the extract to obtain an upper liquid; And

(d) concentrating the upper liquid.

In the step (a), powdering can be carried out after the hoe saengmabjang is washed with fresh water, lyophilized, and then powdered.

In the step (b), water is used as an extraction solvent, and ultrapure water is preferably used, but not limited thereto.

In the step (b), the mixing ratio of the hoe saengmabak powder and water may be 1: 8-12 (W / V). In the embodiment of the present invention, the mixing ratio is 1:10.

In the step (c), centrifugation is preferably performed at 3,000 to 4,000 rpm for 10 to 20 minutes, but is not limited thereto.

The extract prepared by the above-mentioned method can be used after being dried at 30 to 40 < 0 > C and dried.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments and experiments described below in detail. It should be understood, however, that the invention is not limited to the disclosed embodiments and examples but is to be embodied in different forms and should not be construed as limited to the embodiments set forth herein, It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

[ Example ]

<Statistics Processing>

Statistical analysis of all experimental results was performed by SAS software (SAS Institute Inc., Cary, NC, USA) and the results were analyzed by Duncan's multiple range test at p <0.05 Were examined.

Example  One. Hornsey  Mushroom extract manufacturing

Hoe used in this experiment life is Sargassum (Sargassum horneri ) was collected from Busan Metropolitan City and washed with fresh water, lyophilized, powdered, vacuum packed and stored at -20 ℃.

10 times as much ultrapure water was added to the powdered Hornsia sauces and extracted with a shaker (shaker: H-0802, Dongwon science co., Busan, Korea) for 24 hours at room temperature. After centrifugation at 3,000 rpm for 10 minutes using a centrifuge (UNION 32R, Hanil, Incheon, Korea), the supernatant was taken and the residue was extracted twice with 10 times of ultrapure water by the same method. The extracted supernatant was filtered with a filter paper, concentrated using a rotary evaporator (RE200, Yamato Co., Tokyo, Japan) and dried at 37 ° C. The dried samples were stored at -20 ° C and used for the experiments.

Example  2. Mouse preparation and allergen induction

Six-week-old female BALB / c mice were purchased from Orient Co. (Seongnam, Korea) and used in antiallergic experiments. Male BALB / c mice at 4 weeks of age were used in an anti-atopic dermatitis test. The mice were pre-fed for 1 week in an animal room maintained at a temperature of 20 ± 2 ° C, a humidity of 50 ± 10%, and a 12-hour light-dark cycle.

To induce allergy to mice, sensitization was performed by injecting 20 쨉 g / ml ovalbumin (OVA) and 2.6 mg / ml aluminum hydroxide gel into the abdominal cavity of mice and intraperitoneally injecting the same antigen one week later to induce allergy .

Example  3. From allergy-inducing mice Splenocyte  Isolation and Culture

Mice that induced allergy were sacrificed by cervical dislocation, and the spleen was aseptically excised. The spleen was washed with RPMI 1640 medium and homogenized with a tissue grinder to liberate the cells. The cell suspension was centrifuged at 1,800 rpm for 5 minutes at 4 ° C and then allowed to stand for 10 minutes in RPC lysis buffer (Tris-buffered ammonium chloride, 0.87% NH4CL, pH 7.2) to remove red blood cells. Thereafter, PBS, ovalbumin (100 μg / ml) and horseshoe crab (1, 10, 100 μg / ml) were added to the spleen cell suspension diluted to a concentration of 2 × 10 ⁶ cells / ml by adding 10% FBS- / Ml) was added, and the cells were cultured in a CO ₂ incubator (MCO-15AC, Sanyo, Osaka, Japan) at 37 ° C for 72 hours.

Example  4. Hornsey  Evaluation of cytotoxicity of mungbean extracts

The spleen is the main secondary lymphoid organs where an immune response to blood-derived antigens occurs, divided into red and white. The red lobe is composed of red blood cells, dendritic cells and macrophages, and the white lobe consists of T lymphocytes that are responsible for cell mediated immunity and B lymphocytes that are responsible for humoral immunity. In this experiment, we evaluated the cytotoxicity of the extract of Hornbill moth, Cyperus alba, by the effect of the extract of hornbill moth, Cytochrome c, on splenocyte proliferation.

The MTT assay was performed using the spleen cells cultured in Example 3 in order to examine the effect of the extract of the present invention on the proliferation of spleen cells.

As a result, as shown in FIG. 1, at a concentration of 10 μg / ml, there was no significant difference compared to those of the ovalbumin supplement, but the splenocyte proliferative potency was increased by about 230% at a concentration of 100 μg / ml. These results indicate that the extracts of Hornsby's mongolica extract do not have cytotoxicity and that the secretion of IL-4 and IgE is not decreased by apoptosis. In addition, it was found that the extract of hornbill 's yeast extract stimulates splenocyte proliferation at a high concentration of 100 μg / ml.

Experimental Example  One- Hornsey  Inhibitory effect of mungbean extract on allergies

The effect of the extract of the present invention prepared in Example 1 on interferon-gamma (hereinafter abbreviated as 'IFN-γ'), IL-4 and total IgE secretion of allergen-induced mouse spleen cells Respectively.

Experimental Example 1 -1: From allergy-inducing mice Splenocyte  Isolation and Culture

Mice were purchased from Orient Co. (Seongnam, Korea) at a temperature of 20 ± 2 ° C, a humidity of 50 ± 10%, and 1 hour in an animal room where a 12-hour light-dark cycle was maintained After preliminary breeding for week, they were used for the experiment. To induce allergy to mice, sensitization was performed by injecting 20 쨉 g / ml ovalbumin (OVA) and 2.6 mg / ml aluminum hydroxide gel into the abdominal cavity of mice, and intraperitoneal injection of the same antigen a week later induces allergy . The allergen-induced mice were sacrificed by cervical dislocation, and the spleen was aseptically extracted. The spleen was washed with RPMI 1640 medium and homogenized with a tissue grinder to liberate the cells. The cell suspension was centrifuged at 1,800 rpm for 5 minutes at 4 ° C and then allowed to stand for 10 minutes in RPC lysis buffer (Tris-buffered ammonium chloride, 0.87% NH4CL, pH 7.2) to remove red blood cells. Thereafter, PBS, ovalbumin (100 μg / ml) and horseshoe crab (1, 10, 100 μg / ml) were added to the spleen cell suspension diluted to a concentration of 2 × 10 ⁶ cells / ml by adding 10% FBS- / Ml) was added, and the cells were cultured in a CO ₂ incubator (MCO-15AC, Sanyo, Osaka, Japan) at 37 ° C for 72 hours.

Experimental Example 1 -2: Splenocyte Splenocyte IFN -γ, IL -4 measurement of secretion

The supernatant was separated by centrifugation at 10,000 rpm for 3 minutes (Micro 17 RT, Hanil Co., Incheon, Korea), and then ELISA kit (Mouse ELISA set, BD Bioscience, San Diego, USA) And the amount of secretion of INF-y and IL-4 was measured. First, ELISA microplates were coated with anti-mouse IFN-y and IL-4 antibody by aliquot and blocked with 10% FBS solution. (Streptavidin-conjugated streptavidin-horseradish peroxidase conjugated with biotinylated anti-mouse INF-γ, IL-4 antibody, and biotin-conjugated biotinylated anti- Streptavidin horseradish peroxidase conjugate was added. After the OPD solution was added to the reaction mixture, the absorbance was measured at 490 nm using a microplate reader (Model 550, Bio-Rad, Richmond, USA).

As a result, as shown in Fig. 2, IFN-y secretion level was significantly higher than that of the ovalbumin-supplemented group at all the addition concentrations of the extracts supplemented with the extract of hornblende extract. In the case of IL-4 secretion, And it decreased in concentration - dependent manner.

Experimental Example 1 -3: Splenocyte  all ( total ) IgE  Secretion measurement

The total amount of IgE secretion in the splenocyte culture fluid was measured by the same method as in the measurement of cytokine secretion amount of the splenocytes using a total IgE ELISA kit (Mouse ELISA set, BD Bioscience, San Diego, USA).

As a result, as shown in Fig. 3, the total IgE secretion amount of the spleen cell cultures was significantly lower than that of the ovalbumin supplemented group at all the addition concentrations of the extracts of hornblende extract, and the values were similar to those of control.

In conclusion, it was found that the extract of the present invention increases the type 1 cytokine, IFN-y, and effectively reduces the secretion of IL-4 and IgE, the type 2 cytokines. Therefore, the extract of the present invention of the present invention was thought to be effective for inhibiting allergies through the control of cytokine type 1 and type 2.

Experimental Example  2- Hornsey  Inhibitory effect of mung bean extract on atopic dermatitis

Experimental Example 2 -1: Effect of the extract of the present invention on symptoms of atopic dermatitis

A 2,4-dinitrochlorobenzene (DNCB), a hapten forming substance that artificially causes dermatitis, was applied to the back region of BALB / c mice to produce an atopic dermatitis-like mouse model. The effects of hoe saimyung mackerel extract on atopic dermatitis symptoms were investigated.

Male BALB / c mice at 4 weeks of age were used in an atopic dermatitis experiment. The mice were pre-fed for 1 week in an animal room maintained at a temperature of 20 ± 2 ° C, a humidity of 50 ± 10%, and a 12-hour light-dark cycle.

The symptoms of atopic dermatitis were observed immediately after epilating and after 1 week and 2 weeks of application of the extract of hornblende extract of the present invention, appearance changes (erythema, itching and dry skin, edema, erythema and folliculitis) were observed in mouse skin.

As a result, as shown in FIG. 4, there was no difference from the negative control immediately after epilating, and it was confirmed that the symptoms of erythema, scab and dryness were reduced in the group treated with hoe saengmyung mackerel extract for one week as compared with the positive control group I could. In addition, it was confirmed that the erythema symptoms were significantly reduced when the horns were applied for 2 weeks. It is considered that the moisturizing effect of the polysaccharide present in the extract of the present invention reduces the symptoms such as dryness and erythema. The above results show that the extract of hoe saengmyung mackerel reduces the symptoms of atopic dermatitis, It was judged to be available.

Experimental Example 2 -2: Splenocyte IFN -γ, IL -4 measurement of secretion

In order to investigate the inhibition of atopic dermatitis by the Hornsey mash mash extract, the extract of the present invention was applied to mice having atopic dermatitis, and splenocytes were isolated and cultured to obtain IFN-γ The IL-4 content was measured.

Methods for isolating and culturing spleen cells from mice that caused atopy, and methods for measuring secretion amounts of IFN-y and IL-4 in the cultured splenocyte culture fluid were used in the same manner as in Experimental Example 1.

As shown in FIG. 5, the amount of IFN-y secretion was 265.3 pg / mL, which was significantly lower than that of the control group of 804.4 pg / mL. In addition, the amount of IL-4 secretion was 40.67 pg / mL, which was significantly lower than that of the positive control with 78.59 pg / mL. Thus, it was confirmed that the extract of the present invention is effective for inhibiting acute phase and chronic period atopic dermatitis.

Experimental Example 2 -3: total ( total ) IgE  Content measurement

The total IgE content of the serum was measured to examine the effect of the extract of the present invention on the secretion of IgE.

Total IgE content in the serum was measured using an ELISA kit (Mouse ELISA set, BD Bioscience, San Diego, USA). First, ELISA microplates were coated with anti-mouse IgE and blocked with BSA. Biotinylated anti-mouse IgE and biotinylated streptavidin-conjugated streptavidin-horseradish peroxidase conjugated with biotin were added to the diluted serum at a constant concentration and reacted. Streptavidin horseradish peroxidase conjugate was added. The absorbance was measured at 490 nm using a microplate reader (Model 550, Bio-rad, Richmond, USA) after the addition of ortho-phenylenediamine (OPD) solution.

As a result, as shown in FIG. 6, it was found that the total IgE secretion amount of the extract of the present invention was significantly reduced as compared with that of the positive control. This suggests that the secretion of IgE was suppressed by the decrease of IL-4, and the significant decrease of IgE could be suppressed by suppressing the secretion of various inflammatory substances from mast cells and basophils by hornblende extract. This is the result.

Experimental Example 2 -4: in vivo  On Splenocyte Proliferative ability

After spraying the extract of the present invention with the extract of the present invention for 2 weeks on mice induced by atopic dermatitis with DNCB, splenocytes were isolated and cultured for MTT assay.

As a result, as shown in FIG. 7, the splenocyte proliferative capacity of the extract of the present invention was similar to that of the negative control, and about 55% in the positive control. The results showed that the application of DNCB increased the splenocyte proliferative activity to activate the immune cells, whereas the application of the extract of the present invention did not significantly affect splenocyte proliferative activity.

In conclusion, Experimental Example 2 showed that 1) the secretion amount of IFN-γ was increased at a concentration of 1 to 100 μg / ml of the extract of hornbill mushroom of the present invention, and the secretion amount of IL-4 and IgE was significantly reduced, . 2) When the extract of the present invention was applied to a mouse causing atopic dermatitis, symptoms such as erythema and dryness were alleviated. In addition, secretion of IFN-γ decreased by about 60% and secretion of IL-4 decreased by about 50% in spleen cell cultures. The amount of total IgE secreted in the serum after the application of the extract of the present invention was reduced by about 80%. From the above results, it was concluded that the extract of the present invention is effective for the prevention of acute and chronic atopic dermatitis.

Claims (3)

A pharmaceutical composition for the prevention or treatment of atopic dermatitis, which comprises an extract of Hornsby matsuba as an active ingredient.
delete A food composition for preventing or ameliorating atopic dermatitis, which comprises an extract of Hornsby's safflower as an active ingredient.

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