WO2019103571A1 - Composition for treating dermatitis, containing extract and fraction of aerial parts of gypsophila oldhamiana miq, and use thereof - Google Patents

Abstract

The present invention relates to: a composition for treating dermatitis, containing, as an active ingredient, an extract of aerial parts of Gypsophila oldhamiana Miq. or a fraction thereof; and a preparation method therefor.

Description

Compositions for treating dermatitis including fungus over-ground extracts and fractions and uses thereof

To a composition for preventing or treating dermatitis, including dermatan topoisomerase extract, or fraction thereof, and to its use.

The definition of atopic dermatitis (AD) in the medical dictionary is chronic and recurrent inflammatory skin disease, which is characterized by dry and keratinous scaly skin, inflammation, increased skin permeability and susceptibility to common and harmless factors And is vulnerable to surface infections (Ann. Dermatol. 22 (2): 125-137). Typical features include severe redness and itching.

The cause of atopic dermatitis is difficult to explain clearly but it is known that not only genetic factors that cause immune abnormalities but also environmental factors as well as various inducing factors are complicated to develop lesions of atopic dermatitis. Thus, it is clear that atopic dermatitis is a multinational disease caused by the involvement of multiple factors. Especially, it is known that genetic factors are important for the development of atopic dermatitis.

Atopic dermatitis is a clinical manifestation of atopic dermatitis, including allergic diseases such as atopic dermatitis, asthma, allergic rhinitis, and allergic conjunctivitis. The term 'atopic dermatitis' is short and is called 'atopy', but the definitions of the two terms are clearly different medically. Atopic dermatoses tend to be expressed at the same time or at the same time as they interact with each other, so they are described as 'allergy march'. Allergic streaks are the symptoms of atopic dermatitis, food allergies, asthma, and allergic rhinitis in the order of young infants, and are associated with progression of sensitization to allergens. According to domestic studies, the incidence of atopic dermatitis due to genetic factors was 41.7% in parents with atopic disease and 14.7% in parents without allergic disease, indicating a link between atopic dermatitis and parental allergic disease have.

Atopic dermatitis is recognized as the first disease of allergy march and it is used as an index to predict the occurrence of atopic disease. If the immune response is not controlled during the period of atopic dermatitis, it is estimated that the allergic march accelerates, so atopic dermatitis control is necessary to prevent asthma and rhinitis. The pathogenesis of atopic dermatitis is largely divided into an outside-in model associated with skin barrier and an inside-out model associated with abnormal immune response, and an integrated consideration of genetic and environmental factors is required.

At present, the therapeutic agent for atopic disease is focused on steroids and antihistamines which have the effect of inhibiting the secretion of histamine, which is the final product of allergic reaction, or suppressing the inflammatory reaction, which is one of the allergic symptoms, and some immunomodulators and phototherapy are used. Conventional medicines are effective in alleviating allergic symptoms, but they are not a fundamental treatment method, and there is a risk of reducing drug efficacy and various side effects in long-term use. Side effects of steroids include obesity, diabetes, hypertension, and depression. Side effects of antihistamines are depression, concentration problems, lethargy, drowsiness, sexual dysfunction, and side effects of immunosuppressive drugs include local irritation, hypertension, have. In this respect, it is necessary to develop a new concept treatment material which can substitute steroid, antihistamine, immunosuppressant, or use of existing therapeutic agent and thus has less side effects in long-term use.

Interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL-13), which induce T cell hyperactivation, play an important role in the pathogenesis of atopic dermatitis. It is known to be secreted by Th2 cells and plays an important role in the initiation of atopic dermatitis by controlling the synthesis of immunoglobulin E (IgE).

In irritable mice, 2,4, dinitrochlorobenzene (DNCB) and acetone were used to stimulate the back skin, resulting in a delayed allergic reaction in the stimulated local area, resulting in abnormal immune function Skin barrier can be ruined. Thus, a model similar to atopic dermatitis can be created using DNCB to create an in vivo model related to skin barrier function and immune function of the individual. DNCB induces atopic skin dermatitis (TEWL) and pH in mice with increased hydration and decreased hydration.

If DNCB is used to induce lesions such as atopic dermatitis, then skin barrier function, ear thickness, and immunological function can be a very desirable candidate for treating or preventing atopic dermatitis.

Disease is a scientific name Gypsophila Oldhamiana Miq. is a perennial plant belonging to the genus Caryophyllaceae. The height of the plant is 50-100cm, and the stem has branches on the upper part and no hair on the whole. Root leaves are lanceolate, with three leaf veins, sharp ends, no petiole, but narrowed like a petiole. The edges are flat and the leaves in the middle are 6cm long and 5 ~ 10cm wide. Flowers bloom from June to July, white, with many flowers hanging from the tip of the branch and the end of the main stem. Fruit is capsule shaped like a round, and divided into 4 pieces. It is said that the root is used for rooting and the roots are planted for planting.

However, there is no study on the fungal growth of the fungus, and it is not known that the extract of the fungal growth factor inhibits the expression of IL-4 and is effective for atopic dermatitis.

One aspect is that the Gypsophila oldhamiana Miq . ) Composition for preventing or treating dermatitis comprising an extract of the above-ground part or a fraction thereof as an active ingredient.

Another aspect provides a method of making a fungal upper aerial part extract comprising contacting a fungal upper part with water, a C1 to C6 alcohol or a mixture thereof.

Yet another aspect provides a method of preventing or treating dermatitis in an individual comprising administering the composition to an individual in an amount effective to prevent or treat dermatitis.

Yet another aspect provides a method of creaming a skin for moisturizing an individual comprising applying the composition to an individual's skin in an amount effective to ameliorate dermatitis.

The first aspect is that Gypsophila oldhamiana Miq.) as an active ingredient, or a composition for preventing or treating dermatitis comprising an extract of the above-ground part, or a fraction thereof, as an active ingredient.

The above fungus body part may be a leaf, a flower, a stem, a bud or a combination thereof, and may be a dry extract.

The extract may be extracted with a hydrophilic solvent. The hydrophilic solvent may be water, a C1 to C10 alcohol, or a mixture thereof. The alcohol may be a compound having at least one -OH group of C1 to C10, C1 to C6 or C1 to C4. The alcohol may be methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol or a mixture thereof.

The fungal upper layer extract may be crude extract, fraction, or a combination thereof. The crude extract is obtained by bringing the above fungal spore part into contact with an extraction solvent, which does not contain a specific component and is not separated. Said fractions are those obtained by separating a substance containing a specific component from the above-mentioned congealed water. The separation can be carried out by separation using an organic solvent, chromatography or filtration. The chromatography may be ion exchange chromatography, affinity chromatography, size exclusion chromatography, HPLC, or a combination thereof.

The extraction may be a step of incubating the fungal spore part in the extraction solvent. The above fungal spore part may be in the form of fine particles which have been pulverized or pulverized. The fungal spore part may be dried or washed. The incubation may be performed at room temperature to reflux temperature without stirring or stirring. The incubation temperature may be appropriately selected depending on the solvent selected. For example, from room temperature to reflux temperature, from 30 ° C to reflux temperature, from 40 ° C to reflux temperature, from 50 ° C to reflux temperature, from 60 ° C to reflux temperature, or from 65 ° C to reflux temperature. The extraction time can be appropriately selected in accordance with the extraction condition. The extraction time may be 2 to 4 hours, for example 2.5 to 3.5 hours. The extraction solvent may be 5 to 15 times, for example, 5 to 13 times, 5 to 11 times, 8 to 15 times, 8 to 13 times, 8 to 11 times, or 9 to 11 times the fungal upper part. The extraction may be one or more times, for example, 1 to 5 times, 1 to 4 times, 1 to 3 times, 2 to 5 times, or 2 to 4 times. The crude extract obtained can be filtered, concentrated and dried. The drying may be reduced pressure drying.

In one embodiment, the extract may be one extracted under ultrasonic irradiation. The ultrasonic irradiation may be performed at room temperature, 10 to 25 占 폚, 10 to 30 占 폚, 10 to 40 占 폚, 10 to 45 占 폚, 10 to 50 占 폚, 10 to 55 占 폚, 10 to 60 占 폚, Lt; 0 > C. The irradiation time is 10 to 30 minutes. 10 minutes to 1 hour, 10 minutes to 2 hours, 10 minutes to 3 hours, 10 minutes to 4 hours, 10 minutes to 5 hours, 10 minutes to 6 hours, 10 minutes to 9 hours, 10 minutes to 12 hours, 10 minutes To < / RTI > 24 hours. The ultrasonic irradiation may be performed in an airtight container.

In one embodiment, the extract can be further extracted using an organic solvent. The organic solvent may be ethyl acetate, hexane, dichloromethane, butanol, acetone, acetonitrile or a combination thereof. Extraction with the organic solvent may include adding the extract to the organic solvent and incubating. The extraction conditions may be appropriately selected according to the solvent to be selected, and the incubation may be performed at room temperature to reflux temperature without stirring or stirring. The incubation temperature may be appropriately selected depending on the solvent selected and may be selected from the range of, for example, room temperature to reflux temperature, 30 占 폚 to reflux temperature, 40 占 폚 to reflux temperature, 50 占 폚 to reflux temperature, Deg.] C to reflux temperature. The extraction time can be appropriately selected in accordance with the extraction condition. The extraction time may be 2 to 4 hours, for example 2.5 to 3.5 hours. The extraction solvent may be 5 to 15 times, for example, 5 to 13 times, 5 to 11 times, 8 to 15 times, 8 to 13 times, 8 to 11 times, or 9 to 11 times the fungal upper part. The extraction may be one or more times, for example, 1 to 5 times, 1 to 4 times, 1 to 3 times, 2 to 5 times, or 2 to 4 times. The resulting extract can be filtered, concentrated and dried. The drying may be reduced pressure drying.

The extraction may further comprise HPLC of the crude extract or its fractions. The HPLC may be a semi-preparative HPLC.

In one embodiment, the composition may be one comprising as an active ingredient a fraction obtained by fractionating the fungal upper aerial part extract. The fraction is obtained by suspending the above fungal extract in water and extracting it with methylene chloride (CH ₂ Cl ₂ ), ethyl acetate (EtOAc) or butanol ( n- BuOH) as a fractionating solvent, Depending on the fractionating solvent, it may be methylene chloride fraction, ethyl acetate fraction or butanol fraction.

In one embodiment, the above Fusarium aerial part extract or fraction thereof may prevent or prevent dermatitis by inhibiting hyperactivation of immune cells to stabilize or restore immune function or inhibit the expression of interleukin-4 (IL-4) Improvement, or treatment.

The dermatitis may be selected from the group consisting of skin pruritus, seborrheic dermatitis, contact dermatitis, atopic dermatitis, diabetic dermatitis, folliculitis, acne, allergic dermatitis and neurogenic dermatitis.

Thus, the above fungus aerial part extract or fraction thereof in the composition may be contained in an amount effective to inhibit the expression of IL-4 in cells, to improve dermatitis, or particularly to improve atopic dermatitis. The effective amount may be appropriately selected depending on the cell or individual selected by a person skilled in the art. The effective amount is in the range of 0.1 mg to 1,000 mg, such as 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg 5 mg to 5 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg . The composition may be one for inhibiting the expression of IL-4 in cells in vitro or in an individual. Cells in an individual may be located, for example, at a site where dermatitis progresses due to increased expression of IL-4, or at sites where inflammation is excessively advanced by deposition of IL-4 and externally manifests dermatitis symptoms. The dermatitis may be atopic dermatitis. In addition, the dermatitis may be a skin infectious disease caused by weakening of skin barrier function in a patient with atopic dermatitis.

The composition may further comprise a cosmetically, pharmaceutically, or pharmaceutically acceptable excipient or carrier. The composition may be a pharmaceutical, food or cosmetic composition.

The food composition may be used alone or in combination with other food or food ingredients, and may be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the pharmaceutical composition of the present invention may be added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or drink is prepared. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, and includes foods in a conventional sense.

The beverage may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

The food composition may also be used for nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , Or a combination thereof. The food composition may also contain natural fruit juice, fruit juice drinks, flesh for the manufacture of vegetable drinks, or combinations thereof. Such additives may be selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition.

The composition may be formulated into oral or parenteral dosage forms. Oral administration formulations may be granules, powders, solutions, tablets, capsules, dry syrups, or combinations thereof. The oral dosage form may be an injection or an external preparation for skin. The external skin preparation may be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal patch, a drug-containing bandage, a lotion, or a combination thereof.

The external preparation for skin is a composition for external use for skin such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorbent, a whitening agent, an antiseptic, an antioxidant, a surfactant, , Various skin nutrients, and the like can be appropriately compounded as needed.

The external preparation for skin may be a metal blocker such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate or gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridine, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, fructose, fructose and other herbal medicines, various herbal medicines, tocopherol acetate, glycyrrhizic acid, Sugars such as trehalose and the like can also be appropriately compounded.

The composition may comprise a pharmaceutically acceptable diluent or carrier. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low substituted hydroxy cellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, calcium monohydrogen phosphate anhydrous, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or combinations thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The composition may be a cosmetic composition for preventing or treating dermatitis. The cosmetic composition may be used for skin moisturizing. The cosmetic composition may be topically applied to areas without skin inflammation. The cosmetic composition may be used separately from the pharmaceutical composition for treating dermatitis.

The composition may be a food composition for preventing or treating dermatitis.

The second aspect provides a method of making a fungal upper aerobic part extract comprising contacting the fungal upper aerobic part with water, a C1 to C6 alcohol or a mixture thereof.

The contacting may be carried out under ultrasonic irradiation or under reflux. The method may further comprise the step of suspending the extract in water, followed by fractionation with methylene chloride, ethyl acetate, butanol, or a combination thereof.

The third aspect provides a method of preventing or treating dermatitis of an individual comprising administering the composition to a subject.

Administration can be by any method known in the art. Administration may be by any means directly administered to a subject, such as by intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration, . The administration can be systemically or locally administered. The administration may be local administration to a site where dermatitis is present or a site where the skin is dry or prone to dryness. One example of a local administration may be to apply to the skin.

The composition may be administered with other drugs such as steroids, anti-inflammatory agents, antihistamines, antibiotics, and antifungal agents to efficiently improve the dermatitis of an individual. The administration may be sequentially, concurrently or separately administered to a subject.

The subject may be a mammal, such as a person, a cow, a horse, a pig, a dog, a sheep, a goat, or a cat. The subject may be an individual in need of improvement of dermatitis, such as atopic dermatitis.

Such administration may comprise administering the fungal upper aerial part extract or fraction thereof in an amount of from 0.1 mg to 1000 mg, such as from 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1000 mg, 5 mg to 5 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg to 25 mg, 10 mg to 1000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 100 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 50 mg, or 10 mg to 25 mg per day.

The fourth aspect provides a method for creaming a skin for moisturizing an individual comprising applying the composition to an individual's skin in an amount effective to improve dermatitis.

According to a composition for preventing or treating dermatitis in an individual according to one aspect, it can be used for preventing or treating dermatitis such as atopic dermatitis.

According to another aspect of the present invention, it is possible to efficiently produce an extract or a fraction thereof on the ground surface of the fungal spore.

According to another aspect of the method for preventing or treating dermatitis of an individual, the disease of the individual can be effectively prevented or treated.

According to another aspect of the present invention, there is provided a cosmetic method for skin moisturization of an individual, which can efficiently maintain skin moisturization of an individual.

FIG. 1 is a graph showing the degree of inhibition of IL-4 by a ratio of PI-treated group to the effect of IL-4 on the expression of IL-4 on the undifferentiated part and the underground part of extract.

FIG. 2 is a graph showing the results of measurement of transdermal water loss (TEWL) according to time elapsed after treatment with the extracts of the root and upper part of the blightless mouse.

FIG. 3 is a graph showing the results of measurement of moisture content after treatment of shoots and shoots in the hairless mouse. FIG.

FIG. 4 is a graph showing the results of measurement of the amount of IgE protein in serum after treatment with the extracts of the ground and bottom part of the fungus in the hairless mouse.

FIG. 5 is a graph showing the results of measurement of the amount of IL-4 protein in serum after treatment with the extracts of the ground and bottom part of the fungus in the hairless mouse.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Above and below ground extract and his Fraction  Identification of manufacturing and its effects

In this example, the extracts and fractions thereof were prepared, and the effects of the extracts and fractions thereof on the improvement of atopic dermatitis were confirmed.

1. Preparation of fenugreek extract and its fractions

(1) Preparation of fenugreek extract

The ground and underground parts of the fennel used in this example were collected directly from Daejeon-dong, Gangneung, Gangwon-do, Korea, and dried.

(1.1) Preparation of ethanol extract of above ground plant part

500 g of the dorsal root part was cut into a diameter of 0.5 cm or less, soaked in 1.5 L of ethanol in a glass container, and extracted with ultrasonic extraction for 3 hours. The thus-obtained extract was dried under reduced pressure and concentrated to obtain 22.2 g of a concentrate (hereinafter referred to as " fennel ground-ethanol extract ").

(1.2) Preparation of fractions of ethanol extract

(CH ₂ Cl ₂ ), ethyl acetate (EtOAc) and butanol (n-BuOH) were suspended in 50 ml of distilled water according to the systematic fractionation method, and 20.0 g of the above- ), And the mixture was fractionated and concentrated under reduced pressure to obtain 5.6 g of a methylene chloride layer, 3.2 g of an ethyl acetate layer, 3.6 g of a butanol layer, and 3.2 g of a water layer, and the methylene chloride (CH ₂ Cl ₂ ) fraction , Ethyl acetate (EtOAc), butanol (n-BuOH) and water fractions.

(1.3) Preparation of bamboo underground-ethanol extract

The underground part of the dwarf sprout was extracted with 12 kg of 100% ethanol and 1.2 kg of the seaweed root was extracted with ultrasound using an ultrasonic device (ENDELIN SONOREX) for 3 hours. The extract obtained by repeating this three times was dried under reduced pressure and concentrated to obtain 34.7 g of a concentrate (hereinafter referred to as "a sprout-bottom part-ethanol extract").

(1.4) Fruit of the bottom part - ethanol extract Fraction  Produce

34.7 g of the ethanol extract from the bottom part of the dana plant prepared in (1.3) was suspended in 500 ml of distilled water and extracted three times with the same amount of hexane, dichloromethane, ethyl acetate and butanol ( n- BuOH) After fractionation, the mixture was concentrated under reduced pressure to obtain 1.98 g of a hexane layer, 0.77 g of a dichloromethane layer, 0.45 g of an ethyl acetate layer and 4.93 g of a butanol layer to obtain a fraction of methylene chloride (CH ₂ Cl ₂ ), ethyl acetate (EtOAc) (n-BuOH) and water fractions.

2. Fenugreek extract and his The fraction  Effect on IL-4 Expression

The rat basophilic leukemia cells (RBL-2H3 cells) used in this experiment were purchased from Korea Cell Line Bank (Seoul, Korea).

RBL-2H3 cells were inoculated into each well of a 6-well plate at a concentration of ⁵ × 10 ⁵ cells / well and cultured in DMEM containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Dulecco 'Modified Eagle Medium) medium at 37 ° C and 5% CO _2. After 16 hours, 1 μM cyclosporine A and a sample were added at a concentration of 10 μg / ml and cultured for 1 hour Pretreatment. The sample is the fungal upper body extract prepared in Section 1 or its fraction. Next, 50 ng / ml of phorbol 12-myristate 13-acetate (PMA) and 1 μM ionomycin were added and cultured for 9 hours under the same conditions. PMA and ionomycin are used to stimulate intracellular production of cytokines such as interferon, perforin, IL-2, and IL-4.

Cells were harvested from the QIAsymphony or QIAQube (Qiagen) instrument using RNeasy mini kit (Qiagen) to extract RNA. The extracted RNA was checked for integrity using an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, Calif., USA). cDNA was synthesized from cDNA of 1 의 of RNA using ImProm-II Reverse Transcription System (Promega, Madison, Wis., USA). PCR was performed using GeneAmp PCR System 9700 (Applied Biosystems, Foster City, Calif., USA).

The PCR reaction was carried out in the same manner as in Example 1 except that 0.02 μl Ex taq polymerase (TAKARA, Otsu, Shiga, Japan), 2 μl Ex taq polymerase buffer, 1.6 μl dNTP (10 mM), 2 μl forward primer (20 μM), 2 μl reverse primer , 11.38 [mu] l of water and 1 [mu] l of synthesized first strand cDNA were mixed well to perform PCR on the reaction mixture in a total volume of 20 [mu] l.

The PCR conditions for IL-4 were 94 ° C for 4 minutes, 94 ° C for 30 seconds, 60 ° C for 30 seconds, 72 ° C for 30 seconds (25 times) and 72 ° C for 5 minutes (1 time) Beta-actin conditions were 94 ° C for 4 minutes (1 time), 94 ° C for 30 seconds, 55 ° C for 30 seconds, 72 ° C for 30 seconds (20 times) and 72 ° C for 5 minutes (1 time). RT-PCR results were measured using QIAxcel advanced (Qiagen) instruments. The efficacy was determined by comparison with the ratio of [PMA + ionomycin] (PI) treated group to 100. The primers used for RT-PCR are shown in Table 1.

	Forward primer	Reverse primer
IL-4	SEQ ID NO: 1	SEQ ID NO: 2
beta -actin	SEQ ID NO: 3	SEQ ID NO: 4

Table 2 and FIG. 1 show the effect of IL-4 on the expression of IL-4 in the PI treated group as 100, and the inhibition of IL-4 by the ratio thereof.

sample	IL-4 expression (%)
Untreated group	13.51
PI	100
PI + CsA	29.5
PI + ground ethanol extract	31.4
PI + overhead CH 2 Cl 2 fraction	18.9
PI + ground EtOAc fraction	16.2
PI + ground n-BuOH fraction	21.6
PI + underground ethanol extract	35.5
PI + bottom CH 2 Cl 2 fraction	21.4
PI + bottom EtOAc fraction	20.5
PI + underground n-BuOH fraction	15.4

In Figure 1, CsA represents cyclosporin A. As shown in Table 2 and Fig. 1, the extracts and fractions of each extract of fenugreek inhibited the expression of IL-4 in RBL-2H3 cells. Among them, the extracts and fractions of the fungal extracts and fractions significantly inhibited the expression of IL-4 as compared with the underground part. This result shows that the extracts and fractions of the above-ground fruit juices can help alleviate atopic symptoms.

3. Fungal growth of the ground-ethanol extract On DNCB  Effects of atopic dermatitis on skin barrier function in hairless mice

Hairless mice (purchased from Orient Bio) at 6 weeks of age (female, 23-27g) were separated into four groups: untreated group, 2,4-dinitrochlorobenzene (DNCB) induced Vehicle treated group, DNCB induced dendritic surface part, and underground part treated group. DNCB was prepared by mixing 3: 1 by volume of 99% acetone in water with olive oil as a solvent. The vehicle was a mixture of propylene glycol and ethanol in a volume ratio of 7: 3. One (w / v /)% DNCB solution was applied once a day for 200 days to mice and the like for 7 days after the group separation. A total of 7 times 0.1% (w / v)% DNCB solution was applied at intervals of 2 days after one week without treatment for one week after the last application. Apply 1% (w / v)% DNCB solution and 1% (w / v)% solution of ground and bottom extracts with vehicle as solvent for 14 days and vehicle divided into morning and afternoon. Respectively.

The skin moisture content (TEWL, Trans Epidermal Water Loss) was measured using aquaflux (Biox, London, UK) and the skin hydration and pH were measured using Skin-O-Mat COSMOMEP, Berlin, Germany). First, DNCB was measured 7 days, 14 days and 21 days after induction of atopic dermatitis.

The results are shown in Fig. 2 and Fig. FIG. 2 shows the results of measurement of transdermal water loss (TEWL) according to time elapsed after treatment with the extracts of the root and upper part of the shoots. FIG. 3 shows the results of measurement of moisture content after treatment of shoots and shoots in the hairless mouse.

As shown in Fig. 2, it can be seen that the amount of transdermal water loss in the untreated mice was lower than that in the control group, as compared with the untreated group. Therefore, it can be seen that the skin barrier function of the dandelion root extract is significantly enhanced in the uninhabited mice as compared with the vehicle.

In addition, as shown in Fig. 3, it was confirmed that each extract of the pupae increased hydration compared to the control vehicle in the uninhabited mouse. Among them, the water extracts from the above ground water extracts showed higher water content than those from the underground water extracts. Therefore, the extracts of Abelmoschus taurus enhance the skin barrier function in uninjured mice compared to the vehicle and the underground part.

When the skin barrier is broken, the pH value of the skin surface is increased. As a result of measuring the pH value of the skin with time after treatment of each extract of Daesung, The pH value of the hairless mice was not significantly changed compared to the control vehicle. Therefore, it can be seen that the skin barrier function is strengthened by the over-the-ground extract of Daesung.

	0 days	21st
CON	6.475	6.3375
Vehicle	6.5125	6.925
Dandelion	6.525	6.125
Underground	6.55	6.575

The amount of interleukin-4 (IL-4) protein, one of immunoglobulin E (IgE) and cytokine of immune cells, which is one of the most important factors in atopic dermatitis, is used as a marker of atopic dermatitis in blood plasma in blood. . Specifically, after anesthetizing the mouse, blood is collected from the abdominal aorta using a syringe containing 30 μl of 0.18 M EDTA. After centrifugation at 8,000 rpm for 15 minutes at 4 ° C using a centrifuge, plasma and blood cells are separated. A mouse IgE ELISA quantitation kit (Bethy Laboratories, Montgomery, TX, USA) and a mouse IL-4 ELISA quantitation kit (Bethy Laboratories, Montgomery, TX, USA) were used in double plasma.

The results are shown in Fig. 4 and Fig. FIG. 4 shows the results of measurement of the amount of IgE protein in serum after treatment with the extracts of the ground and bottom part of the fungus in the hairless mouse. FIG. 5 shows the results of measuring the amount of IL-4 protein in the serum after treatment with the extracts of the ground and bottom part of the fungus in the hairless mouse.

As shown in Figs. 4 and 5, it was confirmed that the amount of IgE and IL-4 protein in plasma was lower in the fungal extracts than in the control mice in the untreated mice. Among these extracts, the topical extract of Fusarium spp. Significantly reduced the amount of IgE protein in serum compared to other extracts.

Claims (16)

1. Gypsophila oldhamiana Miq.) composition for preventing or treating dermatitis comprising a topical extract, or a fraction thereof, as an active ingredient.
2. The composition according to claim 1, wherein the flesh root portion is dried.
3. The composition of claim 1, wherein the extract is extracted in a hydrophilic solvent.
4. The composition of claim 2, wherein the hydrophilic solvent is water, a C1 to C6 alcohol, or a mixture thereof.
5. 4. The composition of claim 3, wherein the alcohol is ethanol, methanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol or mixtures thereof.
6. The composition of claim 1, wherein the extract is extracted with reflux.
7. The method according to claim 1, wherein the fraction is obtained by suspending the above fungal spore part and underground part in water and extracting it with methylene chloride (CH ₂ Cl ₂ ), ethyl acetate (EtOAc) or butanol ( n- BuOH) Methylene chloride fraction, ethyl acetate fraction, or butanol fraction.
8. The composition according to claim 1, wherein said extract is to stabilize immune function or inhibit IL-4 expression.
9. The composition according to claim 1, wherein the dermatitis is selected from the group consisting of skin pruritus, seborrheic dermatitis, contact dermatitis, atopic dermatitis, diabetic dermatitis, folliculitis, acne, allergic dermatitis and neurogenic dermatitis.
10. The composition of claim 1, further comprising a cosmetically, pharmaceutically, or pharmaceutically acceptable excipient or carrier.
11. The composition according to claim 1, wherein the composition is a pharmaceutical, food or cosmetic composition.
12. The composition of claim 1, wherein the composition is a cosmetic composition for moisturizing the skin.
13. Contacting the persimmon ground part with water, a C1 to C6 alcohol or a mixture thereof under ultrasonic irradiation.
14. 14. The method of claim 13, wherein the method further comprises fractionating the extract in water followed by methylene chloride, ethyl acetate, butanol, or a combination thereof.
15. A method for treating dermatitis of an individual comprising administering to a subject an effective amount of a composition according to any one of claims 1 to 11 for treating dermatitis.
16. A method for creaming the skin of an individual comprising applying the composition of claim 12 to the skin of an individual in an amount effective to improve dermatitis.

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