KR101747499B1 - Composition for preventing skin aging, treating wrinkled skin, preventing and treating dermatitis comprising lindera sericea extract - Google Patents

Abstract

The present invention relates to a composition for prevention of skin aging, improvement of wrinkles and prevention and treatment of skin diseases, which comprises extracting Lindera sericea . More particularly, it relates to a composition for inducing the production of collagen, Which is capable of reducing the secretion of immunoglobulin and histamine, in skin aging prevention, wrinkle improvement, pharmaceutical preparation for skin disease prevention and treatment, and cosmetic and functional food for therapeutic use.

Description

FIELD OF THE INVENTION The present invention relates to a composition for preventing skin aging, improving wrinkles, and preventing and treating skin diseases, comprising a hair extract, a wood extract, a skin extract,

The present invention relates to a composition for prevention of skin aging, improvement of wrinkles, prevention and treatment of skin diseases, which comprises a hair extract, and more particularly to a composition for controlling inflammation and immune diseases, The present invention relates to a novel use of a ginseng extract as a natural material for preventing aging of skin, improving wrinkles, preventing and treating skin diseases, cosmetics and functional foods.

Collagen, ceramide, etc. are known to be factors affecting skin aging. Collagen is the main component of cell epilepsy, and it decreases with age, and when it is 80, collagen decreases to 65% of 20s. The reduction of collagen and ceramide is the main cause of skin wrinkles due to aging, and collagen and proteoglycan cure by intermolecular cross-linking or side chain modification, resulting in changes in cell-extracellular matrix interactions, Is known to occur. It is also known that accumulation of abnormal protein due to intracellular stress of the embryo, reduction of cell hyperplasia, and induction of skin rash caused by decrease of epidermal cell turnover (usually 28 days) are also involved in skin aging. It has been reported that when estrogen is administered to improve the wrinkles of the skin, the increase of skin collagen and the synthesis of hyaluronic acid are improved and the moisturizing property is improved (EL-Domyati M et al .: Exp Dermatol 11, 2002).

Oxidative stress is an important cause of aging. Examples of such stress inducers include reactive oxygen species (ROS) including peroxides, hydroxy radicals, hydrogen peroxide, and the like. In addition, reactive nitrogen species (RNS) such as nitric oxide, peroxynitrite, and lipid peroxides are known as oxidative stress inducers. These oxides are known to cause skin aging and cancer by binding to the DNA base guanine to induce mutation. In addition, these oxides are involved in the arachidonic acid cascade, which may cause vasoconstriction and platelet aggregation, and may develop as a risk factor for arteriosclerosis or Alzheimer's disease.

As active substances capable of inhibiting oxidative damage, water-soluble components such as vitamin C and vitamin B12, liposoluble natural components such as lycopene and beta-carotene are well known. However, there have been few studies on aging regulators using natural materials.

Skin diseases, including atopy, are recurrent chronic dermatitis with severe itching, occurring in about 10 to 15% of children, of which 90% are reported to heal spontaneously within 5 years. Such skin diseases generally improve in adulthood, and about 30 to 40% of patients with childhood illnesses do not recur dermatitis on appearance, while the rest are adult skin irritation, skin irritation by irritant substances, dermatitis such as housewife eczema Is known to suffer difficulties. In addition, in the case of sensitive skin, there is a high possibility that the skin becomes atopic skin because of low water retention and recovery ability, skin keratinization and itching.

Although the exact pathophysiology of skin disease has not yet been fully understood, it is believed that immunologic and nonphysiological mechanisms are involved with genetic predisposition. Most cases of extrinsic skin disease, which accounts for the majority of skin diseases, are caused by immunologic mechanisms associated with immunoglobulin IgE, including delayed immune responses due to abnormalities of T cells rather than immediate immune responses to specific allergens. Recently, Th2-related cytokines such as IL-4, which induce IgE production from B cells, have been reported to cause skin diseases (JS Kang et al., IHhibition of atopic dermatitis by topical application of silymarin in NC / Nga mice. intl immunopharm. (2008) 8 1475-1480).

In order to treat such a skin disease, conventional steroids such as ceramide, linoleic acid, vegetable oil or mineral oil, steroid preparations such as hydrocortisone, or substances enhancing antimicrobial and anti-inflammatory functions, Synthetic inhibitors, cell proliferation inhibitors, inflammation and itching inhibitors have been proposed. However, the above steroid preparations may cause adverse effects such as inhibition of growth of the epidermis or side effects, urea peroxide and the like may cause excessive skin irritation, antibiotics such as antihistamines may cause resistance to bacteria and photosensitizers And the like. In addition, the above steroid preparations may cause serious side effects such as capillary vasodilation and increase in the thickness or expansion of the stratum corneum when applied to the skin for a long period of time. In recent years, gamma-linoleic acid -0046633) is easily oxidized to have poor stability and skin irritation is relatively strong, which makes it difficult to use it for sensitive skin.

Recently, the focus has been on the search for natural substances having an effect on skin diseases. Representative examples of the extracts of mugwort (Korean Patent No. 10-0377262), Ganoderma lucidum, Yugaekipi, Wooly bamboo tree, Baekbongyeong, Baekjima and Baekjyoncho extracts (Korean Patent No. 10-0517465), evening primrose oil, aloe, (Korean Patent No. 10-0451444), celadon lobules (Korean Patent No. 10-0451444), celestial lobules, green tea leaves, green tea leaves, green tea leaves, (Korean Registered Patent No. 10-048439), lavender, eucalyptus and tea tree oil (Korean Patent No. 10-0454752), Kimchi, Uiin, Omija, Korean Patent No. 10-0597997). In addition, it is known that there is an effect of the above-mentioned life, myrrh, genus, and malt. However, there is a disadvantage in that the use of the natural substances is limited because of the effect of the natural substances or the allergic hypersensitivity reaction in the sensitive skin even though they are effective.

Therefore, it is demanded to develop a substance which is more effective and safe than conventional wrinkle improvement and skin disease prevention and treatment agents.

Lindera sericea is a deciduous broad-leaved shrub that grows only in Mt. Mudeung and Jojoe in Chonnam province. Its stem color is light green and straight. It grows well in moist and fertile soil. The leaves are long elliptical, with slanting, with fine hairs on both sides and grayish white on the back. It is dioecious in April, with yellow flowers gathering beneath the freshly bloomed leaves, and the fruit is round and ripens in black in October. Leaves and branches are boiled for tea and used as spices.

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KR 1020000046633 A KR 100377262 B KR 100517465 B KR 100451444 B KR 100454752 B KR 100483539 B KR 100597997 B

It is an object of the present invention to provide a composition for preventing skin aging and improving wrinkles, which comprises a hair follicle extract exhibiting collagen production effect.

It is another object of the present invention to provide a cosmetic for preventing skin aging and wrinkles and a functional food comprising an extract of Fir tree as an active ingredient.

It is another object of the present invention to provide a composition for preventing and treating skin diseases, which comprises an extract of Fusarium oxysporum exhibiting an anti-inflammatory effect and a cytokine secretion-inhibiting effect.

It is another object of the present invention to provide a pharmaceutical preparation and a functional food for preventing and treating skin diseases comprising an extract of Fusarium oxysporum as an active ingredient.

According to one aspect of the present invention, there is provided a composition for preventing skin aging and wrinkles, which comprises extract of Lindera sericea .

In addition, the hair promoting tree may be at least one selected from the group consisting of leaves, flowers, branches, fruits and roots of hair promoting trees.

The hair extract may be an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms.

According to another aspect of the present invention, there is provided a cosmetic for preventing skin aging and wrinkles, which comprises extract of Lindera sericea as an active ingredient.

The cosmetic may also be in the form of a nanoliposome.

According to another aspect of the present invention, there is provided a functional food for preventing skin aging and improving wrinkles, which comprises an extract of Lindera sericea as an active ingredient.

According to another aspect of the present invention, there is provided a composition for preventing and treating skin diseases, which comprises extract of Lindera sericea .

In addition, the hair promoting tree may be at least one selected from the group consisting of leaves, flowers, branches, fruits and roots of hair promoting trees.

The hair extract may be an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms.

According to another aspect of the present invention, there is provided a pharmaceutical preparation for preventing and treating skin diseases, which comprises extract of Lindera sericea .

In addition, the pharmaceutical preparation may include an external preparation for skin.

In addition, the external preparation for skin may be a nanoliposome formulation.

According to another aspect of the present invention, there is provided a functional food for skin disease prevention and improvement, which comprises extract of Lindera sericea as an active ingredient.

A composition comprising a hair fry extract according to one aspect of the present invention may induce collagen production. Thus, it is possible to exert an excellent effect in preventing skin aging and improving wrinkles.

In addition, a composition comprising a hair fry extract according to one aspect of the present invention can inhibit nitric oxide species and cytokines. Thus, it is possible to exert an excellent effect in prevention and treatment of skin diseases.

The composition comprising the hair extract of the present invention according to one aspect of the present invention is excellent in safety to human body and thus can be used for safe cosmetics, medicines or foods.

FIG. 1 shows the results of measurement of cytotoxicity against HDFn cells of the extract of Fusarium oxysporum of the present invention,
FIG. 2 shows the results of measuring the cytotoxicity of RAW 264.7 cells of the hair extract of the present invention,
FIG. 3 shows the results of measuring the collagen synthesis activity of the hair extract of the present invention,
FIG. 4 shows the results of measurement of the nitrogen oxides secretion inhibitory activity of the hair extract of the present invention,
FIG. 5 shows the results of measurement of the IgE secretion inhibiting activity of the extract of Fusarium oxysporum of the present invention,
Fig. 6 shows the results of measurement of the inhibitory activity against histamine secretion of the extract of the fir tree of the present invention.

The present invention relates to a composition for preventing skin aging, improving wrinkles, preventing and treating skin diseases, a cosmetic preparation, a pharmaceutical preparation and a health functional food characterized by containing a extract of Lindera sericea .

Hereinafter, the present invention will be described in more detail.

According to one aspect of the present invention, there is provided a composition comprising a hair fry extract.

The hair promoting wood used in the present invention may be at least one member selected from the group consisting of leaves, flowers, branches, fruits and roots of hair promoting wood, preferably at least one member selected from the group consisting of leaves, Is selected. This can be cut, dried and pulverized by a conventional method.

The method for separating the hair extract from wood is not particularly limited, but a method commonly used in the art can be used for producing the extract. For example, a method using an extraction device such as hot water extraction, immersion extraction, reflux cooling extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction and ultrasonic extraction, or a method using an adsorption resin containing XAD and HP-20 Method, but the present invention is not limited thereto. Preferably, it can be obtained by treatment with an extraction solvent, followed by concentration under reduced pressure.

As the extraction solvent, a polar solvent may be used. The polar solvent may be at least one selected from the group consisting of water, alcohol, acetic acid, dimethyl-formamide (DMFO) and dimethyl sulfoxide (DMSO), but is not limited thereto.

In the present invention, it is preferable that the hair extract is an extract soluble in a polar solvent containing a lower alcohol having 1 to 4 carbon atoms. More preferably, the hair extract of the present invention is an extract which is soluble in a polar solvent containing a lower alcohol having 1 to 2 carbon atoms. The lower alcohol having 1 to 2 carbon atoms may be methanol or ethanol, more preferably ethanol.

The extraction solvent may be added in an amount of 5 to 20 times the total dry weight of the hairless wood. Preferably, it is added at a factor of 20 times the total dry weight of the hairpin.

The temperature at which the extractant is separated by treating the extraction solvent may be 10 to 120 ° C. Preferably, it may be 40 to 100 ° C. The extract may be extracted at the above temperature for 1 to 7 hours to isolate the hair extract of the present invention. Preferably 3 to 5 hours, more preferably 3 hours, to isolate the hair extract of the present invention.

The composition comprising the hair extract of the present invention can produce collagen. Through this, it is possible to prevent aging of the skin and improve the skin wrinkles.

In addition, the composition comprising the hair extract of the present invention can inhibit the release of nitric oxide from the cells. Through this, skin aging can be prevented and an anti-inflammatory effect can be exhibited.

In addition, the composition comprising the hair extract of the present invention can inhibit the secretion of cytokines including immunoglobulin IgE and histamine. Through this, it is possible to exhibit the preventive and therapeutic effect of skin diseases by reducing the immune hypersensitivity reaction. Examples of the skin diseases include, but are not limited to, atopic dermatitis and the like.

Since the hair extract of the present invention does not show toxicity to cells, it can be used as safe medicines and food additives.

The composition comprising the hair extract of the present invention can be prepared into a pharmaceutical preparation. The pharmaceutical preparations of the present invention can be prepared by using at least one selected from the group consisting of pharmaceutically acceptable carriers, diluents and excipients according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by formulating it into a unit dose form or by inserting it into a multi-dose container. The formulations may be oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, or in the form of external preparations, suppositories, sterile injectable solutions, and the like. The formulation may further comprise a dispersant or stabilizer.

If desired, the pharmaceutical preparation may further comprise a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are those conventionally used in the field of the art and include, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium But are not limited to, phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like no.

If desired, the pharmaceutical preparations of the present invention may further contain additives known in the art in addition to the above components. For example, the preparation of the present invention can be administered in various formulations, oral or parenteral, at the time of actual clinical administration, and it can be prepared by further including diluents and excipients in the case of formulation. Examples of the diluent include, but are not limited to, fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. In the case of solid formulation of the present invention, it is preferable to add an excipient such as starch, calcium carbonate, sucrose, lactose, gelatin and the like to the hair extract of the present invention Can be further added. In the case of solid preparation of the hair extract of the present invention, a lubricant such as magnesium stearate, talc and the like may be further added in addition to the excipient.

Liquid form preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. When the hair extract of the present invention is formulated into liquid form, a diluent such as water or liquid paraffin is further added to the hair extract of the present invention, and in addition to the diluent, an excipient such as a wetting agent, a sweetening agent, , A preservative, and the like.

The preparation for parenteral administration may be prepared by adding a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried preparation, a suppository, or the like. Examples of the non-aqueous solvent and suspension include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. For example, witepsol, macrogol, tween 61, cacao paper, laurin, glyceroglatin and the like may be used as the left-handed zero.

The pharmaceutical preparation containing the hair extract of the present invention can be administered orally or parenterally, and in the case of parenteral administration, it can be administered by systemic administration or topical administration. For example, in the case of parenteral administration, it can be administered by a method such as skin application, intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., and preferably, transdermal administration. The appropriate dosage of the pharmaceutical composition comprising the hair extract of the present invention may vary depending on factors such as the formulation method, administration method, condition of the patient, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness Factors, and the ordinarily skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention may be 0.0001 to 1 g / kg, preferably 0.001 to 200 mg / kg. The administration may be carried out once a day or several times in divided doses.

The composition comprising the hair extract of the present invention may be made of a material that is applied to the skin. For example, it can be manufactured as a skin external preparation or cosmetic, which is a pharmaceutical preparation.

The external preparation for skin or cosmetics containing the hair extract of the present invention is preferably a nanoliposome formulation, but is not limited thereto. The nanoliposomes can be mutual spontaneous binding and aligned colloids of particles composed of amphiphilic molecules with water-soluble hair and insoluble tail. The nanoliposome has a bilayer structure similar to a cell membrane, and a composition containing the nanoliposome can exhibit an effect of restoring damaged skin barrier, strengthening skin barrier, and alleviating skin irritation.

The nanoliposomes may comprise saturated or unsaturated fatty acids of alkyl chains having from 12 to 22 carbon atoms, and intercellular lipids. The saturated or unsaturated fatty acid may include, but is not limited to, at least one selected from the group consisting of lauric acid, myristic acid, salicylic acid, stearic acid, oleic acid, linoleic acid and palmitoleic acid. Preferably, it may include at least one selected from the group consisting of stearic acid and palmitoleic acid.

The nano-liposome may have an average particle diameter of 1 to 100 nm, preferably 30 to 70 nm, but is not limited thereto.

The nanoliposome may be contained in an amount of 0.05 to 5.0 wt% based on 100 wt% of the total amount of the external preparation for skin or cosmetics, but is not limited thereto. Preferably, it may be contained in an amount of 0.1 to 4.0 wt%.

The content of the hair extract of the present invention contained in the nanoliposome may be 1.0 to 20.0 wt% based on 100 wt% of the total nanoliposome, but is not limited thereto. Preferably 3.0 to 12.0 wt%.

If desired, the external preparation for skin or cosmetic containing the hair extract of the present invention may further contain additives conventionally used in the art. Examples of the emulsifier include fatty substances, organic solvents, solubilizers, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, perfumes, surfactants, water, ionic emulsifiers, But are not limited to, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic active agents, lipophilic active agents, lipid vesicles and the like. The additive may be included in an amount usually added in the art.

The cosmetic material containing the hair extract of the present invention may be prepared in any form conventionally produced in the art, and examples thereof include a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, , Surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, sprays, and the like. More specifically, the present invention relates to a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizer, a nutrition lotion, a massage cream, a nutritive cream, a moisturizing cream, a hand cream, A cleansing lotion, a cleansing cream, a body lotion, a body cleanser, an emulsion, a press powder, a loose powder, an eye shadow, and the like.

The composition comprising the hair extract of the present invention can be produced as a functional food. The functional food refers to a food prepared by adding a hair extract to a food material, or a food prepared by extracting a hair extract from a capsule, a powder, or a suspension, and has a usual meaning Of healthy foods. Examples of the food include drink, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snack, confection, pizza, noodle, gum, ice cream, soup, beverage, tea, vitamin complex But is not limited thereto.

In the functional food, the content of the hair extract is not particularly limited and may be adjusted to an appropriate range depending on the duration of the ingestion, the purpose of ingestion, and the like. For example, 0.1 to 90 parts by weight, but is not limited thereto.

If necessary, the functional food of the present invention may further comprise at least one selected from the group consisting of carbohydrates and flavors.

The carbohydrate may further include monosaccharides, disaccharides, polysaccharides, sugars and sugar alcohols. But is not limited to, glucose, fructose, maltose, sucrose, dextrin, cyclodextrin, xylitol, sorbitol, erythritol and the like. The content of the carbohydrate may be 1 to 20 parts by weight per 100 parts by weight of the extract of the present invention, but is not limited thereto. Preferably, the content of the carbohydrate may be 5 to 12 parts by weight per 100 parts by weight of the extract of the present invention.

The flavoring agent may further include tau martin, stevia extract, saccharin and aspartame, but is not limited thereto.

If necessary, the functional food of the present invention may further comprise components that are ordinarily added in food production. For example, there may be mentioned nutrients, vitamins, minerals (electrolytes), flavorings, colorants, thickeners, pectic acids, pectate salts, alginic acid, alginates, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, , Carbonating agent, pulp, and the like, but is not limited thereto. The content of the above-mentioned commonly added components is not particularly limited and may be, for example, 0.1 to 20 parts by weight per 100 parts by weight of the extract of Fusarium oxysporum, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to the following examples and comparative examples in order to facilitate understanding of the present invention. It will be apparent to those skilled in the art that these embodiments are illustrative of the present invention and are not intended to limit the scope of the appended claims and that various changes and modifications may be made to the embodiments within the scope and spirit of the invention , And it is natural that such variations and modifications fall within the scope of the appended claims.

Example

≪ Preparation of hair extract and tree extract >

The branches of the hair promoting tree were selected and adjusted according to the degree of pharmacopoeia. Then, 100 g of 70% (v / v) ethanol was added in 2 L of 20 times of the hair growth promoter, After that, the extract was concentrated and dried to obtain a hair extract.

<Cytotoxicity measurement>

Prior to confirming the effect of the extract of the present invention, the cytotoxicity test was carried out to determine a suitable concentration which does not exhibit toxicity to the cells and which can produce an optimal effect without inducing apoptosis. Cytotoxicity of the extracts of hairy root extracts was measured in human dermal fibroblast neonatal (HDFn, Gibco invitrogen cell culture) and rat macrophage RAW 264.7.

HDFn cells are cultivated in a culture medium in _{37 ℃, 5% CO 2 10} % FBS serum, penicillin (100 units / ㎖) , streptomycin (100 ㎍ / ㎖) and 2% is added M106 LSGS (Low SErum Growth Supplement) , And subculture was performed when the cell density of the cell culture container was 80% or more. At the time of subculture, the cells were washed once with SFM (Serum Free Media), treated with trypsin / EDTA (0.05% trypsin, 0.53 mM EDTA) and desorbed at 37 ° C. The cells were recovered by centrifugation (1,300 rpm, 4 ° C) The cells were cultured in the same manner as in Example ¹ , except that the medium was changed at intervals of 2 to 3 days.

RAW 264.7 cells were cultured in Dulbecco's modified Eagle's Medicum (DMEM) containing 10% inactivated fetal bovine serum (GIBCO), 1% penicillin and 1% streptomycin at 37 ° C and 5% CO ₂ Respectively.

Cytotoxicity was measured by MTT assay. The MTT assay is a measurement method that utilizes the principle that the tetrazolium salt is converted to a formazan dye by the mitochondrial dehydrogenase in living cells. The tetrazolium salt added to the medium is converted to the formazan pigment by the succinate-tetrazolium-reductase, which exists in the respiratory chain of mitochondria and is active only in the surviving cell. When the number of living cells increases, the total activity of the mitochondrial dehydrogenase The increase in the activity of this enzyme leads to an increase in the production of formazan pigment, so that the number of cells with metabolic activity in the formazan pigment and medium shows a linear correlation. That is, it can be confirmed that the higher the concentration of formazan pigment, the greater the number of living cells.

HDFn cells were cultured in a 96-well plate at a concentration of 1 × 10 ⁵ cells / well at 37 ° C. and 5% CO ₂ for 24 hours. After removing the culture medium and cultivating the hair extracts at 10, 50 and 100 ㎍ / ㎖ for 24 hours, the medium was removed again and washed twice with PBS (phosphate buffered saline). 100 μL of MTT dissolved in PBS at 300 μg / mL was added and cultured at 37 ° C. and 5% CO ₂ for 4 hours. The culture medium was inhaled and removed, treated with 150 μL of DMSO per well, stirred for 30 minutes, and the absorbance at 570 nm was measured with an ELISA reader. The results are shown in FIG.

Referring to FIG. 1, it can be confirmed that the hair extract of Fusarium wilt does not exhibit cytotoxicity against HDFn cells at all concentrations treated.

For the cytotoxicity of RAW264.7 cells, RAW264.7 cells cultured under the above conditions were mixed at a concentration of 1 × 10 ⁴ cells / well in a 96-well plate and cultured for 24 hours. After the culture medium was removed, the extracts were treated with 10, 50 and 100 ㎍ / ㎖ of the extract, and the cells were cultured for 48 hours. Then, the medium was removed and washed once with PBS. 100 占 퐇 of MTT at a concentration of 0.5 mg / ml was added and cultured at 37 占 폚 and 5% CO ₂ for 4 hours. After the culture medium was inhaled and removed, 200 μL of DMSO was added to each well, and the mixture was stirred in a shaker for 10 minutes. The absorbance at 570 nm was measured with an ELISA reader. The results are shown in FIG.

Referring to FIG. 2, it can be seen that the extract of Fusarium wilt did not show cytotoxicity against RAW264.7 cells at all treated concentrations.

&Lt; Collagen &lt;

Collagen aggregation performance of the hair extract was determined by using the C-terminal recognition antibody of collagen precursor (PIP, Procollagen Type 1 C-peptide).

HDFn cells at a concentration of 5 × 10 ⁴ cells were inoculated into a 48-well plate at 0.3 ml and cultured at 37 ° C and 5% CO ₂ for 24 hours. The hair follicle extracts were diluted to 10, 50 and 100 μg / And cultured for 24 hours. Cells were harvested and the amount of C-terminus of the collagen precursor was measured using an enzymes-linked immunoassay kit (Takara). Collagen concentration was 0 ng to 640 ng (10, 20, 40, 80, 160, 320, 640 ng / mL), and the amount of collagen in the medium was calculated. The results are shown in Fig.

Referring to FIG. 3, it was confirmed that collagen synthesis activity was statistically significant (p value = 0.0068) in the HDFn cells when the hair extract was treated at a concentration of 100 μg / ml.

&Lt; Noxious Oxide Secretion Suppression Experiment >

The inhibition of nitric oxide secretion, an inflammatory factor, was performed on Raw 264.7 cells. Raw 264.7 cells were cultured in a 24-well plate at a concentration of 5 × 10 ⁵ cells / well, and the medium was replaced with SFM for the next day, followed by starvation for 8 hours. After the inoculation, 500 μl / well of SFM was added, and the hair extracts were added at the concentrations of 10, 50 and 100 μg / ml, followed by treatment with 2.5 μl / well of LPS stock solution (100 μg / . The culture supernatant (100 μl) was transferred to a 96-well plate. The same amount of Griess reagent was added and incubated for 30 minutes at room temperature. The degree of color development was confirmed by measuring the absorbance at 540 nm using a microplate reader. The results are shown in Fig.

4, when LPS-treated extracts were treated at a concentration of 50 and 100 μg / ml, the LPS-induced nitric oxide (NO) secretion in RAW264.7 cells was significantly (p value = 0.0033 (50 / / ml), 0.0057 (100 / / ml)).

<IgE secretion inhibition experiment>

The IgE secretion inhibitory ability of the hair extract was determined in U266B1 cell line, which is multiple myeloma cells of human. The U266B1 cell line is a B cell line derived from a number of IgE-secreting individuals among various antibody isotypes. When B cells are differentiated into plasma cells, they secrete IgM among various immunoglobulins. It is known that the IgM is converted into various isotypes by a class switching reaction. It is known that various cytokines secreted from T cells plays an important role in the determination. IgM can be converted into various isotypes such as IgA, IgE, and IgE by the cytokines that are present in the environment. Among them, polysaccharide (LPS) / anti-CD40 and interleukin- LPS and IL-4 were added to the culture medium in U266B1 cell line cultures in order to establish the environment in which B cells secrete IgE. Since IL-4 is known to be stimulated by interleukin-4 and IL-4,

U266B1 cells were cultured in RPMI 1640 medium supplemented with 10% FBS, 1 mM sodium pyruvate, 2 mM L-glutamine, 100 U penicillin and 50 mg / ml streptomycin at 37 ° C and 5% CO ₂ . Cells cultured under the above conditions were treated with cells supplemented with 10 μg / ml of LPS and 5 ng / ml of IL-4, and cultured in a 37 ° C. incubator for 72 hours to stimulate the cells. After 72 hours, the cells were treated with a hair extract of Fusarium oxysporum at a concentration of 10, 50 and 100 / / ml, followed by culturing for 3 days. After 3 days, cells were obtained and the concentration of IgE present in the culture solution was measured using an ELISA (BETHYL). The results are shown in Fig.

Referring to FIG. 5, it can be confirmed that the IgE secretion of B cells was markedly reduced (p value = 0.0020) when the extract of Fusarium wilt was treated at a concentration of 100 μg / ml.

<Histamine secretion inhibition experiment>

Histamine secretion inhibition by hairy root extract was measured in HMC-1 cells, a cell line derived from human mast cells. HMC-1 cells were cultured in IMDM medium supplemented with 10% FBS, 1% penicillin and 1% streptomycin at 37 ° C and 5% CO ₂ . The HMC-1 cells were treated with 10, 50 and 100 μg / ml of hair extract, and 20 nM of Phorbol 12-myristate 13-acetate (Sigma) and 2 mM After the ionomycin (Sigma) was treated for 24 hours, the amount of histamine secreted in the culture medium was measured using a histamine assay kit and an ELISA. The results are shown in FIG.

6, histamine release was markedly remarkable (p value = 0.0028 (50 μg / ml), 0.0070 (100 μg / ml)) when the hair extract was treated at a concentration of 50 μg / ml and 100 μg / Can be confirmed.

The composition containing the extract of the present invention can be used for medicines, foods, cosmetics and the like because of its excellent safety against human body. In addition, collagen production is induced, thereby preventing skin aging and improving skin wrinkles. In addition, it is possible to inhibit nitrogen oxides and cytokines, and thereby, it is possible to exert excellent effects in prevention of skin aging, prevention and treatment of skin diseases.

Claims (13)

A composition for improving skin wrinkles, which comprises extracts of Lindera sericea .
[3] The composition for improving skin wrinkles according to claim 1, wherein the hair growth promoter is at least one selected from the group consisting of leaves, flowers, branches, fruits and roots of hair follicles.
[Claim 2] The composition for improving skin wrinkles according to claim 1, wherein the hair extract is an extract soluble in a polar solvent containing an alcohol having 1 to 4 carbon atoms.
A cosmetic for improving skin wrinkles characterized by containing extract of Lindera sericea as an active ingredient.
The cosmetic for improving skin wrinkles according to claim 4, wherein the cosmetic composition is a nanoliposome formulation. delete delete delete delete delete delete delete delete

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