KR101814389B1 - Cosmetic composition containing new zealand spinach extract - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a first extract, more specifically, a first extract comprising a first extract, wherein the first extract comprises 0.1 to 10% by weight based on the total weight of the composition, Inhibit the production of nitric oxide, tumor necrosis factor alpha (TNF-a), interleukin-6 (IL-6), prostaglandin E2 and thymus and activation-regulated chemokines Which has antioxidative, antiinflammatory and atopic induction-suppressing properties.

Description

[0001] The present invention relates to a cosmetic composition containing a new zealand spinach extract,

The present invention relates to a cosmetic composition containing a first extract, more specifically, a first extract comprising a first extract, wherein the first extract comprises 0.1 to 10% by weight based on the total weight of the composition, Inhibit the production of nitric oxide, tumor necrosis factor alpha (TNF-a), interleukin-6 (IL-6), prostaglandin E2 and thymus and activation-regulated chemokines Which has antioxidative, antiinflammatory and atopic induction-suppressing properties.

Skin is a covering covering the body, which physically and chemically protects the body from the outside world, and performs the biochemical function to metabolism of the whole body. When contaminants such as ultraviolet rays, tobacco smoke, and soot are added to the skin, active oxygen is produced. If the active oxygen is not effectively removed by various protective devices in the body, a series of inflammatory reactions are caused and skin damage is caused . Inflammation reaction reduces the water content of the stratum corneum and makes it dry, causing the elastic fibers of the dermal layer to denature, promoting skin aging. Accordingly, various cosmetic compositions have been developed as shown in the following patent documents in order to alleviate the inflammatory reaction and minimize skin damage.

<Patent Literature>

Patent Publication No. 10-1077824 (Announcement of Oct. 28, 2011) "Cosmetic composition for improving atopy containing sunlight extract and preparation method thereof"

However, conventional compositions using ilite, which is a mica-group mineral belonging to the monoclinic system, do not provide any guarantee of stability by using minerals or the like, and compositions using natural materials extracted from various plants (for example, chlorella Japanese Patent JP 10-36283 using extracts, etc.), there is a problem that the effect of alleviating the inflammatory reaction is deteriorated by placing the point of cell proliferation.

SUMMARY OF THE INVENTION The present invention has been made to solve the above problems,

It is an object of the present invention to provide a cosmetic composition containing an antioxidant activity, anti-inflammation and antioxidant-inhibiting antioxidant extract.

It is also an object of the present invention to provide a method for early-stage cultivation which can artificially cultivate fertilizer having antioxidative activity, anti-inflammatory activity and atopy induction inhibitory effect.

In order to achieve the above object, the present invention is implemented by the following embodiments.

According to one embodiment of the present invention, the cosmetic composition containing the first extract of the present invention comprises the first extract.

According to another embodiment of the present invention, the cosmetic composition containing the first extract according to the present invention is characterized in that the first extract comprises 0.1 to 10% by weight based on the total weight of the cosmetic composition.

According to another embodiment of the present invention, in the cosmetic composition containing the first extract according to the present invention, the first extract is pulverized and extracted with 80% ethanol at room temperature for 48 hours to extract the extract, Concentrated under reduced pressure and lyophilized.

According to another embodiment of the present invention, the cosmetic composition of the present invention contains the Nitrogen monosaccharide extract, TNF-alpha, IL-6, prostaglandin, E2 (Prostaglandin E2), and thymus and activation-regulated chemokine.

According to another embodiment of the present invention, in the cosmetic composition containing the first extract of the present invention according to the present invention, the first herb may be a natural or cultivated first herb.

According to still another embodiment of the present invention, in the cosmetic composition containing the first extract according to the present invention, the cultivated first step may be a step of fermenting a livestock, A salty fermentation broth produced by fermenting the produced liquid, anchovy and menthol, and a nutrient supplemented with rice wine diluted in water.

According to another embodiment of the present invention, a method of growing a seedling for use in a cosmetic composition according to the present invention comprises a seed pretreatment step wherein seeds are seeded in water for a predetermined period of time; A seeding step of seeding the seed treated in the seed pretreatment step into a greenhouse having a soil mixed with sand; And a nutrient supply step of supplying liquid fertilizer, a salt fermentation liquid, a nutrient supplement, and water at regular intervals after the sprouting, after the seeding step.

According to another embodiment of the present invention, in the first step of cultivating the composition for use in the cosmetic composition according to the present invention, the liquid fertilizer produced by fermenting the citrus flower and the immature fruit is sprayed at a cycle of 15 days, The liquid fermentation broth produced by fermenting the wax is sprayed every 15 days, the salty fermentation broth formed by fermenting the anchovy and menthol is sprayed every 7 days, and the nutrient solution diluted in water is sprayed at intervals of 15 days .

According to the present invention, the following effects can be obtained by this embodiment.

The present invention contains an antiprotozoal extract and has an antioxidative activity, anti-inflammatory effect and atopy-inducing inhibitory effect.

In addition, the present invention has an effect of artificially cultivating Echinochloa crus-galli which has antioxidative activity, anti-inflammatory activity and atopy induction inhibitory effect.

1 is a table showing the antioxidant activity efficacy evaluation according to Example 2. Fig.
FIG. 2 is a graph showing the evaluation of the inhibitory effect of nitric oxide formation according to Example 3. FIG.
3 is a graph showing the evaluation of the inhibitory effect of TNF-α production according to Example 5. Fig.
4 is a graph showing the evaluation of the inhibitory effect of IL-6 production according to Example 5. Fig.
5 is a graph showing the evaluation of the inhibitory effect of Prostaglandin E2 production according to Example 6. Fig.
FIG. 6 is a graph showing the evaluation of the inhibitory effect on chemokine production according to Example 7. FIG.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the cosmetic composition containing the first extract of the present invention and the cultivation method of the first preparations will be described in detail with reference to the accompanying drawings. Unless defined otherwise, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs and, if conflict with the meaning of the terms used herein, It follows the definition used in the specification. Further, the detailed description of known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted. Throughout the specification, when an element is referred to as "including " an element, it is understood that the element may include other elements as well, without departing from the other elements unless specifically stated otherwise.

The cosmetic composition according to one embodiment of the present invention includes a pre-emergence extract. Demidovia tetragonoides PALL, which is used in the present invention, is a perennial plant in the sandy area of the beach. It grows up to a height of about 50 cm with a small dot on the whole body, but shoots a few branches. Are rhombic close to the egg shape, pointed at the tip of the egg, have no sawtooth on the edge of the leaf, are flat, and one or two of the flowers are attached to the leaf axil without any peduncle, The bell-shaped calyx is like a petal, and the diameter of the flower blooming in yellow is around 6mm. It is known to grow in the sandy beaches and rock slopes of the beach. The cosmetic composition according to the present invention contains an antioxidant activity, an anti-inflammatory activity and an anti-atopy effect. In the first step, cultivated naturally grown and / or artificially cultivated cultivars may be used, and the cultivation method of first crops will be described in detail below.

On the other hand, the first step extract according to the present invention can be prepared as follows. The dried prey is pulverized by a grinding machine and immersed in 80% ethanol for 48 hours. The extract is concentrated under reduced pressure and lyophilized to prepare a preliminary extract.

It is preferable that the progeny early extract is contained in an amount of 0.1 to 10% by weight based on the total weight of the cosmetic composition.

The progeny extract according to the present invention is used for inhibiting the production of antioxidant activity, anti-inflammation and atopy inducing factors. This fact will be described in detail in the following examples.

The cosmetic composition of the present invention may further comprise a composition selected from the group consisting of water-soluble vitamins, oiliness vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

As the water-soluble vitamin, any of vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C and vitamin H can be used, And salts thereof (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbic acid-2-phosphate, magnesium ascorbic acid-2-phosphate etc.) can also be used as water-soluble vitamins . The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzymatic method, or a chemical synthesis method.

As the above-mentioned useful vitamins, any vitamins A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) , And derivatives thereof (such as palmitic acid ascorbin, stearic acid ascorbic acid, dipalmitic acid ascorbic acid, dl-alpha tocopheryl acetate, dl-alpha tocopherol nicotinic acid vitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, Ethyl ether, etc.) and the like are also included in the usable vitamins used in the present invention. The usable vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of a microorganism, an enzyme or a chemical synthesis method.

The polymeric peptide may be any compound as long as it can be compounded in a cosmetic, and preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like. The polymeric peptide can be obtained by a conventional method such as purification from a culture broth of a microorganism, an enzymatic method, or a chemical synthesis method, or purified from natural products such as ducks such as pigs and cows and silk fiber of silkworms .

The polymeric polysaccharide may be any compound as long as it can be compounded in cosmetics, and examples thereof include hydroxyethylcellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.). For example, chondroitin sulfate or a salt thereof can be usually purified from mammals or fish.

The sphingogly lipid may be any as long as it can be compounded in cosmetics, and preferable examples thereof include ceramides, phytosphingosine and sphingoglycolipids. The sphingogly lipids can be purified from ordinary mammals, fish, shellfish, yeast or plants by a conventional method or can be obtained by chemical synthesis.

The seaweed extract may be any of those which can be compounded in cosmetics, but preferably include algae extract, red tiger extract, green algae extract and the like, and furthermore, coloring guanine, arginic acid, sodium alginate , Potassium alginate, and the like are also included in the seaweed extract used in the present invention. The seaweed extract can be obtained from seaweed by a conventional method.

The cosmetic composition of the present invention may contain other ingredients usually added to the cosmetic composition. For example, it is possible to use various additives such as a preservative, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a plant extract, a pH adjuster, Cold agents, antiperspirants, purified water, and the like.

Examples of the above-mentioned fat-soluble ingredients include ester-based fats, hydrocarbon-based fats, silicon-based fats, fluorine-based fats, animal fats and plant fats and the like. Examples of the ester-based oil include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearyl isostearate, But are not limited to, ethylbutyl, butylbutyl, ethylbutylbutyl, butylbutyl, isobutylbutyl, isobutylbutyl, isobutylisobutyl, isobutylisobutyl, isobutylisobutyl, Trimethylol propane, triisostearic acid trimethylol propane, tetra-2-ethylhexanoic acid pentaerythritol tri (2-ethylhexanoic acid) But are not limited to, tallow, caprylic estersyl, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, myristine sasyl, stearyl stearate, decyl oleate, Examples of the isocyanates include isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, isophorone diisostearate, Ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol , Propylene glycol dicaprylylate, dicapric acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated acid glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyl Dodecylsuccinic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, Polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, octyldecyl lactate, octyldecyl isostearate, polyglycerin oleic acid ester, But are not limited to, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, Propyl stearate, propyl stearate, propyl stearate, stearyl stearate, stearyl stearate, propyl stearate, stearyl stearate, Stearyl hydroxystearate, stearyl 12-stearoyl hydroxystearate, 12-stearoyl hydroxystearic acid, Allo one hydroxy stearic acid and the like esters such as cetearyl source. Examples of the hydrocarbonaceous oil include hydrocarbons such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, floating isoparaffin, polybutene, microcrystalline wax and vaseline. Examples of the silicone-based oil include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethyl polysiloxane, decamethyl polysiloxane, dodecamethyl cyclosiloxane, dimethyl siloxane methyl cetyloxysiloxane copolymer, dimethyl siloxane methyl stearoxysiloxane copolymer, Alkyl-modified silicone oil, and amino-modified silicone oil. Examples of the fluorine-based oil include perfluoropolyether and the like. Examples of the animal or plant oil include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, The present invention relates to a process for producing a starch-containing starch, which is selected from the group consisting of seed oil, cottonseed oil, palm oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, Animal or vegetable fats such as oil, can-pick wax, carnauba wax, liquid lanolin, and hardened castor oil.

Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and oil-soluble polymers. Examples of the water-soluble low molecular weight moisturizing agent include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B Propylene glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt and the like. Examples of the lipid-soluble low-molecular moisturizing agent include cholesterol, cholesterol ester and the like. Examples of the water-soluble polymer include carboxyvinyl polymer, polyaspartic acid salt, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, . Examples of the oil-soluble polymer include polyvinylpyrrolidone / eicosene copolymer, polyvinylpyrrolidone / hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

Examples of the molient agent include long chain acyl glutamic acid cholesteryl ester, hydroxystearic acid cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid and lanolin fatty acid cholesteryl ester.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants. Examples of the nonionic surfactant include self emulsifying monostearic acid glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene), sorbitan fatty acid ester, POE sorbit fatty acid ester POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POEPOP (polyoxyethylene polyoxypropylene) copolymer, POEPOP alkyl ether, polyether modified silicone, lauric acid alkanolamide , Alkylamine oxides, hydrogenated soybean phospholipids, and the like. Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylallylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl ginseng salt, alkyl Amide phosphate, alkyloyl taurine salt, N-acyl amino acid salt, POE alkyl ether carboxylate, alkylsulfosuccinate, sodium alkylsulfoacetate, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester and the like . Examples of the cationic surfactant include alkyl trimethyl ammonium chloride, stearyl trimethyl ammonium chloride, stearyl trimethyl ammonium bromide, cetostearyl trimethyl ammonium chloride, distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl ammonium chloride, behenyl trimethyl ammonium bromide, Benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, quaternary ammonium salts of lanolin derivatives, and the like. Examples of the amphoteric surfactant include carboxybetaine, amidebetaine, sulfobetaine, hydroxysulfobetaine, amidosulfobetaine, phosphobetaine, aminocarboxylate, imidazoline derivative, amideamine, etc. , And the like.

Examples of the organic and inorganic pigments include silicic acid, anhydrous silicic acid, magnesium silicate, talc, sericite, mica, kaolin, spinach, clay, bentonite, titanium coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, Inorganic pigments such as aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, chromium oxide, chromium oxide, chromium oxide, But are not limited to, polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluororesin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, Silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and composite pigments of inorganic pigments and organic pigments thereof.

Examples of the organic powder include metal soaps such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; Such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylidene, N-alpha-paratyylnitine, N-alpha-lauroyl arginine, Acyl basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid, alpha-aminoaurauric acid, and the like; Polyethylene, polypropylene, nylon, polymethylmethacrylate, polystyrene, divinylbenzene-styrene copolymer, ethylene tetrafluoride, and the like.

Examples of the ultraviolet absorber include ultraviolet absorbers such as p-aminobenzoic acid, ethyl p-aminobenzoate, amyl paranobenzoate, octyl p-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, benzyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, Benzyl, 2-ethoxyethyl methoxycinnamate, octyl methoxycinnamate, mono-2-ethylhexane glyceryl dipyrromethoxycinnamate, isopropyl methoxycinnamate, diisopropyl-diisopropyl cinnamate ester mixture, Dihydroxymethoxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid, dihydroxybenzophenone sulfonic acid and its salt, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenone disulfonate, dihydroxybenzo Phenanone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianylino-p- (carbo-2'-ethylhexyl- 1,3,5-triazine, 2- (2- And the like can be de-hydroxy-5-methylphenyl) benzotriazole.

Examples of the bactericide include antibiotics such as hinokitiol, trichloroacid, trichlorohydroxydiphenyl ether, crohexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, No. 301, mononitro and eicoll sodium, undecylenic acid, and the like.

Examples of the antioxidant include butylhydroxyanisole, gallic acid propyl, and eicosorbic acid.

Examples of the pH adjuster include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogenphosphate and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, any of the above components may be blended within the range not impairing the object and effect of the present invention, but is preferably 0.01 to 5% by weight based on the total weight, More preferably 0.01 to 3% by weight.

The cosmetic composition of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients conventionally used in cosmetic compositions in addition to the above-mentioned compounds, for example, conventional additives such as stabilizers, solubilizers, vitamins, And a carrier.

The cosmetic composition of the present invention can be prepared in any formulations conventionally produced in the art, and examples thereof include emulsions, creams, foundations, lotions, essences, and hair cosmetic compositions. Specifically, the cosmetic composition of the present invention can be used as a skin lotion, a skin softener, a moisturizing lotion, a nutritional lotion, a massage cream, a nutrition cream, a hand cream, a moisturizing cream, an essence, a pack, a soap, a lotion, a milk lotion, Cleansing foams, body cleansers, astringents, and spray formulations. When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component . When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether. In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters. When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used. When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

Another embodiment of the present invention includes a first step cultivation method. The seedling pretreatment step wherein the first seedling is dipped in water for 24 to 48 hours; A seeding step of seeding the seed treated in the seed pretreatment step into a greenhouse having a soil mixed with sand; A nutrient supply step of supplying liquid fertilizer, salt fermentation broth, nutrient supplement, and water at regular intervals after the first seeding step after the seeding step; And a collecting step of collecting the first step of growing at least three or four months after the nutrient supplying step.

In the seed pretreatment step, the seeds are immersed in the water at room temperature for 24 to 48 hours. In the seeding step, the seeds are planted in a plastic house having soil containing sand for interception with the outside, The juice produced by the fermentation of the citrus flower and the immature fruit was sprayed every 15 days, the juice produced by the fermentation of the wind and the mist was sprayed every 15 days, and the fermented saline produced by fermenting the anchovy and the persimmon was sprayed every 7 days And the nutrient solution diluted in water is sprayed every 15 days. In the cultivation method, conventional liquid cultivation methods used for artificial cultivation of plants can be additionally used. In the first cultivation method, except for the liquid fertilizer, the salt fermentation liquid, the nutrient supplement, and the application period thereof, It is usually followed by the cultivation environment used for the cultivation of plants in vinyl houses. The method of cultivation of the first genera may include a seedling step in which seeds are sprayed on the gravestone and the seeds are transferred into a greenhouse after seeding instead of the seeding step.

In the case of Korea, there are many limitations on the growth environment, and in Korea, it is grown only in certain regions of Jeju Island and the Daedo Sea at a certain time, and it is difficult to cultivate artificially, and artificially cultivated first- There is a difficult economic situation. However, the cultivation method of the present invention allows the cultivation method to obtain the antioxidant activity, the anti-inflammatory activity and the inhibitory effect on the production of the atopy-inducing factor over the whole year regardless of the geographical environment, do. The antioxidative activity, antiinflammation and inhibition of the production of atopy inducing factors of artificially cultivated cultivated plants are described in detail in the following Examples.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these are only for the purpose of illustrating the present invention in more detail, but the scope of the present invention is not limited thereto.

In the following examples, the antioxidative, antiinflammatory, and atopy-inducing factor-producing inhibitory effects of cultivated cultivars cultivated in the first and second cultivation methods were examined. In order to confirm the antioxidative activity, DPPH radical scavenging activity (TNF-α and IL-6), inhibition of prostaglandin E2 (PGE2) production, inhibition of Nitric oxide formation, evaluation of cytotoxicity (LDH assay), proinflammatory cytokines To evaluate the efficacy evaluation and to confirm the inhibitory effect of atopy inducing factor formation, an inhibitory effect of chemokine production (chemokine) production and an assay for cytotoxicity (LDH assay) are performed. The RAW 264.7 cell, which is a murine macrophage cell line used in the following examples, was obtained from the ATCC (American Type Culture Collection, VA, USA) and the HaCaT cell, which is a human keratinocyte cell line, It was used from the department of medicine and pharmacology.

<Example 1> Preparation of a sample of the first extract

The extracts were concentrated under reduced pressure and lyophilized to give a crude extract (sample 1 (sample 1)). The extracts were dried in a hot air drier and then pulverized with a grinder at 80 ° C for 48 hours. ) And cultivated first herb extract (sample 2 (sample 2)). The first step is to use the first step of picking on the shore near the Ole 8 course in Haeyeong-dong, Jeju Special Self-Governing Province, Jeju Special Self-Governing Province in late May.

<Example 2> Evaluation of antioxidative activity by DPPH radical scavenging activity

1) Antioxidant activity was measured by the Blois method, which measures the radical scavenging effect of 1,1-diphenyl-2-picrylhydrazyl (DPPH).

2) Dissolve about 2 mg of DPPH in 15 mL of ethanol to prepare a DPPH solution. DMSO (6.25 mL) is added to 12 mL of this solution, and UV-Vis of the control group is observed at 517 nm. It was diluted with ethanol to an absorbance of 0.94-0.97 and shaken for 10 seconds. Then, 1 mg of each of Samples 1 and 2 was mixed with 1 ml of the solvent, and the resulting solution was sufficiently dissolved. Then, 50 μL of the sample solution was added to 450 μL of the prepared DPPH, and the mixture was allowed to stand at room temperature for 10 minutes and absorbance was measured at 517 nm. The concentration (IC50) of the sample when the absorbance of DPPH decreased by 50% was indicated. Each sample was repeated three times to obtain an average value, and the results are shown in Table 1.

3) Referring to FIG. 1, the IC50 value of the DPPH radical scavenging inhibitory activity of the sample is 450.03 ug / mL in the case of the sample 1 (no pre-fermentation) and 1000 ug / mL or more in the sample 2 And the growth promoting agent of the present invention have an antioxidative effect.

&Lt; Example 3 > Evaluation of anti-inflammatory efficacy (Nitric oxide formation inhibitory effect evaluation)

1) The efficacy of two early extracts, a sample of NO production, which is one of active oxygen and is known to play an important role in inflammation induction, was evaluated.

2) RAW 264.7 cells were adjusted to 1.5 × 10 5 cells / mL using DMEM medium supplemented with 10% FBS and inoculated on a 24-well plate. LPS (1 μg / mL) The amount of produced NO Griess reagent [1% (w / v) sulfanilamide, 0.1% (w / v) naphylethylenediamine in 2.5% (v / v) phosphoric acid] by using the NO ₂ present in the cell culture ^- in the form of Respectively. 100 μL of the cell culture supernatant and 100 μL of the Griess reagent were mixed and reacted on 96-well plates for 10 minutes, and the absorbance was measured at 540 nm. The amount of NO produced was compared with sodium nitrite (NaNO ₂ ) as a standard, and the results are shown in FIG.

3) As shown in FIG. 2, in comparison with the control group, LPS alone treatment, samples 1 and 2 showed a concentration-dependent inhibitory effect on NO production at a concentration of 400 ug / mL or less.

<Example 4> Evaluation of anti-inflammatory efficacy (LDH assay)

1) RAW 264.7 (1.5 × 10 5 cells / mL) was cultured in DMEM medium for 24 hours at the same time as LPS (1 μg / mL) and centrifuged at 3,000 rpm for 5 minutes. LDH (lactate dehydrogenase) assay was performed using a non-radioactive cytotoxicity assay kit (Promega). 50 μl of the culture medium obtained by centrifugation on a 96-well plate and 50 μl of the reconstituted substrate mix were reacted at room temperature for 30 minutes, (Bio-TEK Instruments Inc., Vermont, WI, USA) was used to measure the absorbance at 490 nm. The average absorbance values for each sample group were determined and compared with the absorbance values of the control (LDH control, 1: 5000) to evaluate cytotoxicity.

2) LDH is an enzyme present in the cytoplasm of all cells and catalyzes the reversible conversion between pyruvic acid and lactic acid. When LDH-containing tissue is destroyed, LDH is released into the blood, Both of the two extracts showed no cytotoxicity at concentrations below 400 ug / mL.

<Example 5> Evaluation of anti-inflammatory effect (Evaluation of proinflammatory cytokines (TNF-α and IL-6) inhibitory activity)

1) RAW 264.7 cells (1.5 × 105 cells / mL) were inoculated into 24-well plates using DMEM medium and cultured in a 5% CO2 incubator for 18 hours. Subsequently, the medium was removed and a fresh medium containing 10-fold concentration (1 mg / mL) and a stimulant LPS (1 ug / mL) were simultaneously treated and cultured under the same conditions as the pre-culture. After 24 hours, the culture medium was centrifuged (12,000 rpm, 3 minutes) and the content of pro-inflammatory cytokines produced in the supernatant was measured. The results are shown in FIGS. 3 and 4. All samples were stored at -20 ° C or less until quantification and quantification of pro-inflammatory cytokines was quantitated using a mouse enzyme-linked immunosorbent assay (ELISA) kit (R & D Systems Inc., Minneapolis, MN, USA) The r2 value of the curve was 0.99 or more.

2) Referring to FIGS. 3 and 4, both Sample 1 and Sample 2 showed inhibitory effect on TNF-α and IL-6 production at a concentration of 400 ug / mL or less, and particularly, IL-6 was effectively inhibited.

&Lt; Example 6 > Evaluation of anti-inflammatory efficacy (Evaluation of inhibitory effect of Prostaglandin E2 (PGE2) production)

1) RAW 264.7 cells were adjusted to 1.0 × 10 5 cells / mL using DMEM medium and then inoculated into 24-well plates and cultured in a 5% CO2 incubator for 18 hours. Subsequently, the medium was removed and a new medium containing 50 μL of a 10-fold concentration (1 mg / mL) sample and 450 μL of LPS (1 μg / mL) were simultaneously treated and cultured under the same conditions as the pre-culture. After 24 hours, the culture medium was centrifuged (12,000 rpm, 3 min) to obtain prostaglandin E2 (PGE2). The PGE2 assay was quantified using a PGE2 ELISA kit (R & D Systems Inc., Minneapolis, MN, USA). The results are shown in FIG. 5, and the r2 value of the standard curve for the standard was 0.99 or more.

2) Referring to FIG. 5, Sample 1 showed prostaglandin E2 production inhibitory effect at a concentration of 400 ug / mL, and Sample 2 inhibited the production of prostaglandin E2.

Example 7 Evaluation of Inhibitory Effect on Atopy Inducing Factor (Evaluation of Inhibitory Effect on Chemokine Formation)

1) HaCaT cells (3.0 × 105 cells / mL) were inoculated into 24-well plates using DMEM medium and cultured for 18 hours in a 5% CO2 incubator. After the medium was removed, 50 μL of the sample prepared at 10 × concentration (1 mg / mL) and a new medium containing 450 μL of IFN-γ (10 ng / mL) and 450 μL of TNF-α Lt; / RTI &gt; After 24 hours, the culture medium was centrifuged (12,000 rpm, 3 minutes) to obtain a supernatant. Quantitation of chemokine (TARC), an atopy inducer, was quantitated using an enzyme-linked immunosorbent assay (ELISA) kit (R & D Systems Inc., Minneapolis, MN, USA) The results are shown in Fig. 6, and the r2 value of the standard curve for the standard was 0.99 or more.

2) keratinocytes are involved in the immune response of various skin diseases through interaction with T lymphocytes through various cytokines and cell adhesion molecules. These responses are activated by inflammatory cytokines, such as IFN-γ, secreted by T lymphocytes, and recently, IFN-γ has been reported to increase TARC and MDC expression, known as immunoreactivity factors, in keratinocytes. In the modified keratinocyte HaCaT cells, chemokine was induced with IFN-γ (10 μg / mL) and TNF-α (10 ng / mL) sample 2 inhibited TARC production in a concentration dependent manner at 400 ug / mL or less.

&Lt; Example 8 > Evaluation of inhibitory effect on atopy inducing factor formation (cytotoxicity evaluation (LDH assay)) [

1) HaCaT cells (3.0 × 10 5 cells / mL) were co-treated with DMEM medium and IFN-γ (10 μg / mL) and TNF-α And centrifuged at 3,000 rpm for 5 minutes. LDH (lactate dehydrogenase) assay was performed using a non-radioactive cytotoxicity assay kit (Promega). 50 μl of the culture medium obtained by centrifugation on a 96-well plate and 50 μl of the reconstituted substrate mix were reacted at room temperature for 30 minutes, (Bio-TEK Instruments Inc., Vermont, WI, USA) was used to measure the absorbance at 490 nm. The average absorbance values for each sample group were determined and compared with the absorbance values of the control (LDH control, 1: 5000) to evaluate cytotoxicity.

2) In both HaCaT cells, neither of the two early extracts tested showed cytotoxicity at concentrations below 400 ug / mL.

From the results of the above experiment, it can be seen that the prey has the antioxidative activity, anti-inflammation and the effect of inhibiting the production of atopy-inducing substances. In the case of cultivated cultivars grown by the cultivation method of the present invention, Inflammation and atopy-inducing substance production inhibitory effect.

While the present invention has been described in connection with what is presently considered to be practical exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, Should be interpreted as belonging to the scope.

Claims (8)

delete delete delete delete delete delete delete A seed pretreatment step in which the seeds are soaked in water for a predetermined period of time; A seeding step of seeding the seed treated in the seed pretreatment step into a greenhouse having a soil mixed with sand; And a nutrient supply step of supplying liquid fertilizer, salt fermentation broth, nutrient supplement, and water at regular intervals after the first sprouting after the seeding step,
In the nutrient supply step, the juice produced by fermenting the citrus flower and the immature juice is sprayed every 15 days, the juice produced by fermenting the wind and the mist is sprayed every 15 days, the fermented juice Wherein the nutrient replenisher is sprayed every seven days and the nutrient solution diluted in water is sprayed every 15 days.

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