KR101742029B1 - Composition for preventing skin anti-aging and skin anti-wrinkle comprising a resveratrol-germinated brown rice - Google Patents
Composition for preventing skin anti-aging and skin anti-wrinkle comprising a resveratrol-germinated brown rice Download PDFInfo
- Publication number
- KR101742029B1 KR101742029B1 KR1020150158412A KR20150158412A KR101742029B1 KR 101742029 B1 KR101742029 B1 KR 101742029B1 KR 1020150158412 A KR1020150158412 A KR 1020150158412A KR 20150158412 A KR20150158412 A KR 20150158412A KR 101742029 B1 KR101742029 B1 KR 101742029B1
- Authority
- KR
- South Korea
- Prior art keywords
- brown rice
- resveratrol
- germinated brown
- composition
- skin
- Prior art date
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A23V2200/00—Function of food ingredients
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Abstract
The present invention relates to a composition comprising resveratrol biosynthesis germinated brown rice as an active ingredient. Particularly, the present invention relates to a method for inhibiting the activity of MMP-1, an enzyme involved in collagen degradation, and a method for treating skin aging comprising resveratrol biosynthetic germinated brown rice, which has an effect of increasing the activity of type I procollagen, And to a pharmaceutical composition for improving or treating skin wrinkles. The composition comprising the resveratrol biosynthesis germinated brown rice of the present invention as an active ingredient can be usefully used in various products including pharmaceutical composition, food composition, quasi-drug composition and skin whitening cosmetic composition for preventing or treating skin aging and skin wrinkles
Description
The present invention relates to a composition comprising resveratrol biosynthetic germinated brown rice extract or a fraction thereof as an active ingredient. The present invention relates to a pharmaceutical composition, a food composition, a quasi-drug composition and a skin whitening cosmetic composition for skin aging and skin wrinkle improvement or treatment comprising resveratrol germinated brown rice extract or fraction thereof as an active ingredient.
Collagen, a major constituent of the extracellular matrix, is a major substrate protein produced by fibroblasts in the skin. It is present in extracellular epilepsy. It is an important protein that occupies about 30% of the total weight of bioproteins, and has a solid triple helix structure. Especially, it is high in bone and skin (dermis), and it exists as a fibrous inclusion body in most other body structures. The main functions of collagen are known to be mechanical durability of the skin, resistance of connective tissue and binding force of tissue, support of cell adhesion, induction of cell division and differentiation. Such collagen is reduced by aging and photoaging by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin.
Conventionally, in order to utilize the wrinkle-reducing effect of collagen, products containing collagen in a composition for external application for skin such as cosmetics or ointment have been marketed. However, these products apply collagen itself to the skin surface, It was not able to exhibit the essential wrinkle-improving effect. To solve this problem, there has been a growing interest in collagen synthesis promoting substances. Examples of known collagen synthesis promoting substances include vitamin C, retinoic acid, transforming growth factor (TGF), animal placenta-derived proteins -231370), betulinic acid (JP8-208424), chlorella extract (JP9-40523, JP10-36283, fibroblast proliferation promoting action) and the like. However, there is a problem in that the use amount of the substance is limited or its effect is insufficient due to safety problems such as irritation and redness in skin application, so that a wrinkle reduction effect can not be expected substantially. Therefore, it is urgently required to develop a novel wrinkle-improving composition which is safer to the living body than the conventional wrinkle-improving composition and has a high wrinkle-reducing effect.
As an example of a plant containing a large amount of functional ingredients, rice is widely distributed throughout the world as a one-year-old plant belonging to rice and rice. Rice constitutes 70 to 85% of carbohydrates, 6.5 to 8.0% of protein, 1.0 to 2.0% of fat, and about half of the world's population is used as a stock. Rice is a very important crop grown in about 120 countries, accounting for about 21 of the calories consumed by humans. In the case of brown rice, protein, lipid, fiber, ash and so on are distributed in the chaff and colophony. It is known that the content of minerals and vitamins is 2 to 3 times as high as that of nutrition.
Under these circumstances, the inventors of the present invention conducted studies to investigate the effect of resveratrol biosynthesis germinated brown rice extract on skin aging and skin wrinkle improvement. As a result, resveratrol biosynthesis germinated brown rice extract according to the present invention showed MMP-1 (Procollagen) involved in collagen synthesis, thereby inhibiting photoaging induced by ultraviolet light (UVB), and thus it can be effectively used as an effective ingredient for skin aging and skin wrinkle improvement Thereby completing the present invention.
It is an object of the present invention to provide a pharmaceutical composition for improving skin aging and skin wrinkles, which comprises resveratrol biosynthetic germinated brown rice extract or a fraction thereof as an active ingredient.
Another object of the present invention is to provide a cosmetic composition for improving skin aging and skin wrinkles comprising an extract of germinated brown rice, or a fraction thereof, as an active ingredient.
It is still another object of the present invention to provide a food composition for skin aging and skin wrinkles containing resveratrol germinated brown rice extract or fractions thereof as an active ingredient.
It is still another object of the present invention to provide a quasi-drug composition for skin aging and skin wrinkle improvement comprising an extract of germinated brown rice, or a fraction thereof, as an active ingredient.
In one aspect of the present invention, the present invention provides a composition comprising resveratrol biosynthetic germinated brown rice extract or a fraction thereof as an active ingredient.
The term "resveratrol" used in the present invention is a polyphenolic substance having a strong antioxidative property which is produced when a plant is exposed to a bad environment such as mold or pest, and is known to be contained in excessive amounts in grape skin, grape seed, have. There have been many studies on the effects of resveratrol on humans, and the effects of anticancer, antiviral, neuroprotection, anti-aging, anti-inflammation, and prolongation of life have been known. Resveratrol is there is a cis form (cis) and trans (trans) form has been supposed that the only effect is trans-resveratrol.
As used herein, the term "rice" refers to a plant that is cultivated as an edible crop, particularly in Asia, as a major food resource of more than half of the world's population. Among the rice plants, it means that they are cultivated as grains. There are 20 to 30 kinds of wild species in rice, while the two cultivars are Oryza sativa and Oryza It is known that only two species of glabemina . In the present invention, rice may be a natural type rice or a transformed rice. Specifically, it may be a transformed rice of Dongjinbyeon or Dongjinbyeon which is developed and supplied in Korea. Also, rice seeds containing resveratrol developed in Korean Patent No. 10-0894148 or Korean Patent Laid-open No. 10-2014-0088492 can be used.
The term "rice seeds" used in the present invention includes all states, whether or not rice seeds, rice seeds, and rice bran are included. However, in order to produce germinated brown rice, it may preferably be in the form of brown rice.
The term "resveratrol biosynthesis germinated brown rice" used in the present invention means a form in which rice seeds biosynthesizing resveratrol are ground in a brown rice state and then germinated. Natural rice paddies or transgenic rice seeds, but are not limited to, those which contain resveratrol and are rice seeds in germinated brown rice.
In the present invention, resveratrol biosynthesis sprouting comprises: a) immersing seeds of rice which are biosynthesizing resveratrol in water; And b) germinating the rice seeds immersed in the water by repeatedly treating the rice seeds under the light condition and the dark condition, but the present invention is not limited thereto.
The term "transformation" used in the present invention means all actions in which a specific trait occurs, disappears or is controlled through mutation such as insertion, deletion or substitution on the DNA base sequence. In particular, Which means that the genetic properties of the organism are changed. The term "transgenic rice" used in the present invention means a rice in which DNA base sequences are mutated as compared with natural type rice produced by transformation, and the insertion, modification, or activity of a specific gene And a genetic recombinant produced by mutation of the genetic recombinant.
The term "Dong Jinbin," used in the present invention refers to a breed produced in 1975 by three-hybrid crossing of Nakdongbyeo and Sado Minori, which are resistant to gold dust and disease in Korea in 1975 with the aim of fostering high quality, In 1981, it was decided to be an encouraging variety in Korea.
In the present invention, the resveratrol biosynthetic germinated brown rice extract includes those extracted with water, a lower alcohol having 1 to 4 carbon atoms and a mixed solvent thereof, specifically, methanol, ethanol or butanol. In addition, in the present invention, the extract includes any one of the extract obtained by the extraction treatment, the diluted solution of the extract, the concentrate, the dried product obtained by drying the extract, or the adjusted product or the purified product. For example, the solvent may be 80% methanol, but is not now limited.
In the present invention, resveratrol biosynthesis germinated brown rice extract comprises i) finishing resveratrol biosynthesis germinated brown rice; And ii) adding the ground sprouted brown rice flour to a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms and a mixed solvent thereof, and ii) The step may be repeated two or more times, but is not limited thereto.
In a specific embodiment of the present invention, resveratrol biosynthesis germinated brown rice was ground using a grinding machine in order to prepare the resveratrol biosynthesis germinated brown rice extract of the present invention, and 100 g of the ground germinated brown rice powder was quantified and added to 1 L of 80% And extracted for 30 minutes. After extraction, the supernatant was filtered using a filter paper, and 1 L of 80% methanol was again added thereto, followed by ultrasonic extraction for 30 minutes. The supernatant was filtered again using a filter paper, and the filtrate was collected and concentrated by using a vacuum concentrator. The concentrate was lyophilized to obtain a germinated brown rice extract sample. As a control, germinated brown rice of Dongjin rice was prepared in the same manner as above, and extract samples were prepared by the same method.
In the present invention, a composition containing resveratrol biosynthesis germinated brown rice extract as an active ingredient is characterized in that the biosynthesis germinated brown rice extract is prepared by: (a) immersing the resveratrol biosynthesized rice seed in water; B) germinating the rice seeds immersed in the water by repeatedly treating the rice seeds under light and dark conditions; C) grinding the germinated rice seeds; And d) adding the ground germinated brown rice flour to a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms and a mixed solvent thereof, and extracting the same, Do not.
The term "fraction " as used in the present invention means a product obtained by performing fractionation to separate a specific component or a specific component group from a mixture containing various various components.
The fractionation method for obtaining the fraction in the present invention is not particularly limited and may be carried out according to a method commonly used in the art. A solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a constant molecular weight and a cut-off value, and various chromatography (methods for separating by size, charge, hydrophobicity or affinity , And a combination thereof, and the like. In one embodiment of the present invention, a method of obtaining a fraction from the extract by treating the extract obtained by extracting resveratrol germinated brown rice with a predetermined solvent was used. The kind of the solvent used for obtaining the fraction in the present invention is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water and alcohol: nonpolar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. When alcohol is used in the fraction solvent, C 1 to C 4 alcohols can be specifically used.
For example, the fraction may be prepared by fractionating a resveratrol germinated brown rice extract with a solvent selected from the group consisting of water, an alcohol having 1 to 4 carbon atoms, hexane, ethyl acetate, and a mixed solvent thereof. In another example, the fraction may be prepared by fractionating resveratrol biosynthesis germinated brown rice extract with water, hexane, ethyl acetate or butanol.
Another aspect of the present invention provides a pharmaceutical composition for improving skin aging and skin wrinkles containing the resveratrol germinated brown rice extract or fraction thereof as an active ingredient of the present invention.
In the present invention, resveratrol, resveratrol biosynthesis germinated brown rice and the like are the same as described above.
The pharmaceutical composition of the present invention is a composition having at least one of the following features i), ii) and iii);
i) inhibition of MMP-1 activity,
ii) increased activity of type I procollagen,
iii) increase the thickness of the collagen layer, and may be to improve or treat skin aging and skin wrinkles upon UV (UVB) irradiation.
The active ingredient contained in the pharmaceutical composition of the present invention includes all kinds of substances extracted from the rice such as processed leaves, extracts, fractions and the like to activate active ingredients in resveratrol biosynthetic germinated brown rice.
The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.
As used herein, the term "prevention" refers to any act that inhibits or delays skin aging and skin wrinkles using the composition of the present invention. As used herein, the term "treatment" or "improvement" refers to any action that improves or alters the skin aging and wrinkles of the skin using the composition of the present invention.
The pharmaceutical compositions of the present invention may be formulated into pharmaceutical formulations using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. In the preparation of the formulations, it is preferred that the active ingredient is mixed with or diluted with the carrier, or enclosed in a carrier in the form of a container.
Therefore, the pharmaceutical composition of the present invention can be formulated in the form of an oral preparation, an external preparation, a suppository and a sterile injection solution in the form of powders, granules, tablets, syrups, suspensions, emulsions, syrups and aerosols And may further comprise suitable carriers, formers and diluents conventionally used in the preparation of the composition.
For example, carriers that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.
Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch, calcium carbonate, sucrose, sucrose), lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
Examples of the liquid preparation for oral use include suspensions, solutions, emulsions and syrups, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used have.
Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
On the other hand, ultraviolet B (UVB, ultraviolet-B) has the greatest effect on the skin, mainly penetrating the epidermis and affecting the skin cells, thereby reducing collagen. It is believed that this is a visible reaction of the body by light damage caused by ultraviolet rays.
In the present invention, resveratrol biosynthetic germinated brown rice extract inhibits the activity of MMP-1 involved in collagen degradation, increases the activity of type I prcollagen involved in collagen synthesis, and promotes skin aging and skin wrinkles induced by ultraviolet rays There is an excellent effect of improving.
The term "skin aging" in the present invention means symptoms such as reduction in elasticity of skin, reduction in gloss, generation of wrinkles, weakening of regenerative power, or severe drying, and may be caused by time or external environment.
In the present invention, "preventing or improving skin aging" refers to inhibiting or inhibiting the aging symptom, or alleviating the symptoms of aging already shown.
In the present invention, the term "skin wrinkle " means a skin formed by decay of skin, which may be caused by a cause of a gene, a decrease in collagen existing in the skin dermis, or an external environment.
In the present invention, "prevention or improvement of skin wrinkles" refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating already formed wrinkles.
The major function of dermal fibroblasts is extracellular matrix synthesis and regeneration of injured skin tissue through proliferation. The factors of extrinsic aging such as photoaging, accumulation of damage by free oxygen, and endogenous aging by cell aging due to telomere breakage Reduce the physiological activity of dermal fibroblasts in the dermal layer. Type I collagen, which accounts for more than 95% of the extracellular matrix and plays a major role in the physiological activity of fibroblasts through interactions and signal interactions with adhesion molecules and cytoskeletons of cells, Is reduced, the amount of collagen in the skin tissue is reduced, whereby the surface tension is reduced, so that the skin elasticity is lowered and the skin wrinkles are formed, and the physiological activity including cell proliferation and regeneration is decreased This causes a vicious circle. Therefore, collagen of skin fibroblasts may be directly related to restoration or regeneration of tissue during skin regeneration, skin elasticity, wrinkle formation and skin damage.
That is, when the synthesis of collagen or procollagen of dermal fibroblast is promoted, improvement of skin elasticity, skin regeneration, improvement of skin wrinkles, wound healing, restoration and regeneration of damaged skin tissue, And prevention of skin aging can be obtained.
Accordingly, the composition comprising the resveratrol biosynthetic germinated brown rice of the present invention inhibits collagen degradation and facilitates collagen synthesis, thereby increasing skin elasticity, regenerating skin, improving skin wrinkles, healing wounds, restoring and regenerating damaged skin tissue, And prevention of skin aging and the like. Specifically, prevention of skin wrinkles or skin aging; Or an effect of improvement.
In one embodiment of the present invention, the treatment of resveratrol biosynthesis germinated brown rice extract in photoaging cells induced by ultraviolet irradiation reduces the expression amount of MMP-1 protein involved in collagen degradation and protein expression of type I procollagne involved in collagen synthesis (Fig. 3). It was confirmed that decomposition of the collagen layer was prevented by treatment with resveratrol biosynthesis germinated brown rice extract in artificial skin irradiated with ultraviolet rays (Fig. 4). In addition, the thickness of collagen layer of the mouse treated with resveratrol biosynthesis germinated brown rice for 3 months was 327.8 ± 13.4 袖 m, and the collagen layer of the mouse group fed with Dongjin germinated brown rice for 3 months was 203.3 賊 7.5 袖 m and the mice fed with resveratrol for 3 months The thickness of the collagen layer increased by 1.6 times or more as compared with the collagen layer of 204.4 ± 10.2 μm. In addition, the thickness of the collagen layer of the mouse treated with resveratrol biosynthesis germinated brown rice for 6 months was 444.4 ± 34.1 μm, It was confirmed that the thickness of the collagen layer was increased by 1.9 times or more as compared with the collagen layer 227.8 ± 13.4 μm of the mice fed 6 months and 222.4 ± 11.1 μm of the collagen layer of mice given 6 months long term feeding of resveratrol. As a result, it was confirmed that the resveratrol biosynthetic germination of the present invention not only increases the collagen layer but also suppresses collagen degradation by a light source such as ultraviolet light, thereby preventing skin aging and skin wrinkles.
In another aspect, the present invention provides a cosmetic composition for improving skin aging and skin wrinkles, which comprises an extract of germinated brown rice or a fraction thereof of resveratrol biosynthesis as an active ingredient.
In the present invention, resveratrol, resveratrol biosynthesis germinated brown rice and the like are the same as those described above.
The cosmetic composition according to the present invention can be used as a cosmetic composition in the form of a solution, an ointment for external use, a cream, a foam, a nutritional lotion, a softening water, a pack, a soft water, an emulsion, a makeup base, But are not limited to, emulsions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays no.
In addition, the cosmetic composition of the present invention may further comprise at least one cosmetically acceptable carrier mixed with a cosmetic composition for general skin. Examples of the cosmetic composition include ordinary ingredients such as oil, water, a surfactant, a moisturizer, , A thickener, a chelating agent, a coloring agent, an antiseptic, an incense burner, and the like, but is not limited thereto.
The cosmetically acceptable carrier to be contained in the cosmetic composition of the present invention varies depending on the formulations. When the formulation of the present invention is an ointment, a paste, a cream or a gel, the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide Mixtures of these may be used.
When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as the carrier component, Propellants such as chlorofluorohydrocarbons, propane / butane or dimethyl ether.
When the formulation of the present invention is a solution or emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, and in particular fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.
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 suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
When the formulation of the present invention is a soap, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, a glycerol, have.
The composition of the present invention may further comprise at least one substance having a skin aging function and a skin wrinkle improving function.
Another aspect of the present invention provides a food composition for preventing or improving skin aging and skin wrinkles comprising the resveratrol germinated brown rice extract or fraction thereof of the present invention as an active ingredient.
In the present invention, resveratrol, resveratrol biosynthesis germinated brown rice and the like are the same as described above.
Foods containing the resveratrol biosynthesis germinated brown rice extract of the present invention include, for example, various foods, beverages, gums, tea, vitamin complex, and the like, but are not limited thereto.
The skin aging and skin wrinkle preventing and improving food of the present invention include forms such as tablets, capsules, pills, and liquid preparations.
The beverage composition for skin aging and skin wrinkle prevention and improvement according to the present invention is not particularly limited to the other ingredients except that it contains the resveratrol biosynthetic germinated brown rice extract as an essential ingredient and may contain various flavors or natural carbohydrates And may be contained as an additional component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides, such as maltose, sucrose, etc .; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. (Taurine, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) are advantageously used as flavorings other than the above- . The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 mL of the composition of the present invention.
In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
In another aspect, the present invention provides a quasi-drug composition for prevention or improvement of skin aging and skin wrinkles comprising resveratrol germinated brown rice extract or fraction thereof as an active ingredient.
In the present invention, resveratrol, resveratrol biosynthesis germinated brown rice and the like are as described above.
The term "quasi-drug" used in the present invention means a fiber, a rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a human or an animal, Not belonging to the same category as or similar to those of the machinery or the machine, or as a preparation for sterilization, insecticidal and similar uses for the prevention of infectious diseases, as a diagnostic, treatment, relief, Means goods, machinery or equipment other than those which are not machinery, machinery or equipment of a commodity used for the purpose of prevention and which are used for the purpose of giving pharmacological effects to the structure and function of humans or animals;
The quasi-drug composition may be a disinfectant cleaner, a shower foam, a gagrin, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment or a filter filler. In addition, the quasi-drug composition may include external preparations for skin and personal hygiene products. The external preparation for skin may be prepared and used preferably in the form of an ointment, a lotion, a spray, a patch, a cream, a powder, a suspension, a gel or a gel, But are not limited to, soap, cosmetics, wet tissues, tissue paper, shampoo, skin creams, face creams, toothpaste, lipstick, perfume, make-up, foundation, ball touch, mascara, eye shadow, sunscreen lotion, , Air freshner gel, cleaning gel, and the like.
The composition comprising the resveratrol biosynthetic germinated brown rice extract according to the present invention inhibits the activity of MMP-1, an enzyme involved in collagen degradation, and increases the activity of type I procollagen, an enzyme involved in collagen synthesis, And can be usefully used in pharmaceutical compositions, food compositions, quasi-drug compositions for preventing and treating wrinkles, and cosmetic compositions for improving skin wrinkles.
FIG. 1A shows normal human dermal fibroblast (NHDF) irradiated with 144 mJ / cm 2 of UVB at 10 ug / mL and 100 ug / L of germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol, (NC), UV irradiation group, NR group, UV irradiation group, RR group, RR, UV irradiation-resveratrol biosynthesis group, UV irradiation group, and UV irradiation group. Germinated brown rice treated group, R: UV irradiation - commercial resveratrol reagent treated group).
FIG. 1B is a graph showing the results of a comparison between the NHDF cell group irradiated with 144 mJ / cm 2 of ultraviolet (UVB) at a concentration of 10 ug / mL and 100 ug / L treated with resveratrol biosynthesis germinated brown rice and resveratrol, (NR: treated with Dongjin germinated brown rice, RR: treated with resveratrol biosynthetic germinated brown rice, Resv: commercially available resveratrol reagent group).
FIG. 2A is a graph showing the secretion amount of Matrix metalloproteinase-1 (MMP-1) in UV-irradiated -NHDF cells treated with Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol (NR; RR; resveratrol biosynthetic germinated brown rice group, Resv; commercially available resveratrol reagent group).
2B is a graph showing the secretion amount of type I procollagen in UV-irradiated -NHDF cells treated with Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol (NR; Dongjin germinated brown rice treated group, RR: resveratrol biosynthetic germinated brown rice Treated group, Resv; commercially available resveratrol reagent group).
FIG. 3 is a graph showing the amount of protein expression of MMP-1 and type I procollagen in UV-irradiated -NHDF cells treated with Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol (NR; Dongjin germinated brown rice treated group, RR; resveratrol biosynthetic germinated brown rice group, Resv; commercially available resveratrol reagent group).
4 is a photograph showing the results of ultraviolet irradiation after treating the artificial skin with germinated brown rice germinated and resveratrol biosynthesis germinated brown rice (UVB-; UV-irradiated group, UVB + group, UVB + with NR; UVB + with RR after irradiation with germinated brown rice, UVB + with RR; UV irradiation group after resveratrol biosynthesis germinated brown rice, HE; staining with hematoxylin-eosin (H & E) masson's trichrome) staining).
FIG. 5 is a photograph showing immunostaining of protein expression levels of MMP-1 and type I procollagen after treatment of artificial skin with germinated brown rice and resveratrol biosynthesis germinated brown rice (UVB-; UVB + with NR, UVB + with NR, UVB + with RR, UVB + after UV treatment + UVB + with NRB, UVB + with NR, hematoxylin-eosin, H & E staining, MT, masson's trichrome staining).
FIG. 6 is a photograph showing the analysis of mouse skin tissue collagen layer fed with resveratrol biosynthetic germinated brown rice for 3 months and 6 months (Ctrl: control group (corn feed), DJ; Dongjin germinated brown rice group; Resveratrol group, RR; resveratrol biosynthetic germinated group).
7 is a photograph showing tissue analysis of resveratrol biosynthetic germinated brown rice long-term feeding mice (Ctrl: control group (maize feed), DJ; Dongjin germinated brown rice group, R: resveratrol treated group, RR: resveratrol biosynthetic germinated brown rice group ).
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the embodiments described below. The embodiments of the present invention are provided by way of example to facilitate a specific understanding of the present invention.
Example One: Resveratrol Biosynthetic Germinated brown rice And an extract preparation method
Example 1-1: Resveratrol Biosynthesis Germinated brown rice Produce
The resveratrol biosynthetic rice seeds of the present invention were germinated through the method of Korean Patent Laid-Open No. 10-2011-0136093 to prepare germinated brown rice of the present invention.
Specifically, the filter paper is placed in a Petri dish, water is sufficiently impregnated, resveratrol biosynthesized rice seeds are seeded, and the seeds are cultured in a tissue culture room for 21 to 27, 14 to 18 hours, Germinated for 1 to 7 days. Water was replaced daily to prevent sprouting seeds from rotting.
Example 1-2: Resveratrol Biosynthesis Germinated brown rice Extract preparation
The resveratrol biosynthesis germinated brown rice extract of the present invention was obtained by grinding the resveratrol biosynthesis germinated brown rice of Example 1-1 using a grinder, measuring the germinated brown rice flour at 100 g, adding it to 1 liter of 80% methanol, and extracting it for 30 minutes. After the extraction, the supernatant was filtered using a filter paper, and 1 L of 80% methanol was again added thereto, followed by ultrasonic extraction for 30 minutes. The supernatant was filtered again using a filter paper, and the filtrate was collected and concentrated using a vacuum concentrator. The concentrate was lyophilized to obtain a germinated brown rice extract sample. The amount of resveratrol biosynthesis germinated brown rice used in the present invention represents the amount of resveratrol biosynthesis germinated brown rice extract and the resveratrol content thereof is 0.246 ug / mg as determined by HPLC analysis (Taek Hwan Lee, Biomol Ther 22 (5), 431-437 (2014), Resveratrol-Enriched Rice Down-Regulates Melanin Synthesis in UVB-Induced Guinea Pigs Epidermal Skin Tissue).
As a control group, seeds of Dongjin rice, which is a rice seed to be transformed, were germinated in the same manner as above to prepare germinated brown rice of the control group. Through the same method as the method of preparing the extract samples, a control extract group, Dongjin germinated brown rice extract .
Example 2: ultraviolet ( UVB ) Survey-induced NHDF Assessment of cellular toxicity
Example 2-1: Cell culture
Normal human dermal fibroblast (NHDF) was obtained from a skin biopsy of a healthy male donor (MCTT Core, Inc., Seoul, Korea) and inoculated into a 100 nm tissue culture dish And cultured in DMEM medium containing 10% Fetal Bovine Serum (FBS) and 1% penicillin-streptomycin at 37 ° C and 5% CO 2 . Cells were used between 6 and 10 passages. When cell density reached 80%, seeds were seeded at 1.2 × 10 5 cells / well in a 60 mm culture dish.
Example 2-2: Ultraviolet ( UVB ) Investigation and sample treatment
When the NHDF cells seeded in the 60 mm culture dish reached 80% cell density, the cells were washed twice with PBS (phosphate buffered saline), and then irradiated with ultraviolet light (Bio-Link BLX-312; Vilber Lourmat GmbH, France ) Was irradiated with ultraviolet light of 144 mJ / cm 2 . After UV irradiation, the cells were washed three times with PBS and replaced with a medium containing no serum. Then, the cells were treated with 10 g / mL and 100 g / mL, respectively, of ginseng germinated brown rice, resveratrol biosynthetic germinated brown rice extract and resveratrol .
Example 2-3: Evaluation of cytotoxicity
In order to evaluate the cytotoxicity of the above-mentioned Dongjin germinated brown rice, resveratrol biosynthesis germinated brown rice extract and resveratrol, cell viability was analyzed by MTT analysis and the shape of the pouch was observed through a microscope (FIG. For MTT analysis, the cells were incubated for 72 hours, treated with 100 μL of 1 mg / mL MTT reagent and absorbance at OD 570 nm was measured using an ELISA reader (Molecular Devices E09090; San Francisco, CA, USA) . As shown in FIGS. 1 and 2A, when treated with 10 g / mL and 100 g / mL of the germinated brown rice and resveratrol biosynthetic germinated brown rice, NHDF cells irradiated with ultraviolet light did not show cytotoxicity, It did not change. Resveratrol alone did not show cytotoxicity at the concentration of 10 ug / mL, but showed cytotoxicity at the concentration of 100 ug / mL, and the morphology of the cells changed.
Example 3: Analysis of secretion of enzyme related to skin aging and skin wrinkles
Example 3-1: MMP -1 secretion analysis
Regarding skin wrinkles, MMP-1, an enzyme involved in collagen degradation, and type I procollagen, which is involved in collagen synthesis, are known. NHDF cells were cultured for 72 hours in order to evaluate the ability to regulate MMP-1 and type I procollagen of the above-mentioned Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol, and the conditioned medium was recovered from each well and subjected to ELISA MMP-1 and type I procollagen were measured using a Procollagen Type I C-Peptide EIA Kit (Takara, Shiga, Japan) The secretion level was analyzed. The experiment was repeated three times in the same manner (Figs. 2A and 2B). As shown in FIGS. 2A and 2B, when NHDF cells were irradiated with ultraviolet light, the amount of secretion of 526% MMP-1 was increased compared to cells not irradiated with ultraviolet light. Therefore, it was confirmed that the secretion amount of MMP-1 was decreased in Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol treated group, and compared with the treated rice germinated brown rice group, the amount of MMP-1 secreted in the resveratrol- . Since the resveratrol contained in the resveratrol biosynthesis germinated
Example 3-2: type I procollagen's Secretion analysis
In the secretion control of type I procollagen, it was confirmed that the secretion amount of type I procollagen was reduced by ultraviolet irradiation. Therefore, it was confirmed that the secretion amount of type I procollagen was increased in a dose dependent manner in Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol treated group. In addition, it was confirmed that the resveratrol biosynthesis germinated brown rice had a better effect of increasing the secretion amount of type I procollagen even though the resveratrol content of the brown rice was 0.245 ug / mL corresponding to 1/40 (Fig. 2b).
As a result, it was confirmed that the resveratrol biosynthetic germinated brown rice extract significantly reduced the secretion amount of MMP-1 and increased the secretion amount of type I procollagen in photoaging cells induced by ultraviolet irradiation.
Example 4: Analysis of Expression of Enzyme Proteins Related to Skin Aging and Skin Wrinkle
In order to analyze the protein expression levels of MMP-1 and type I procollagen in UV-irradiated NHDF cells by resveratrol, resveratrol biosynthetic germinated brown rice and resveratrol, Western blotting method was used to analyze the protein content of MMP-1 and type I procollagen The expression level was examined. As shown in FIG. 3, it was confirmed that the amount of protein expression of MMP-1 was reduced in Dongjin germinated brown rice, resveratrol biosynthesis germinated brown rice and resveratrol treated group, and resveratrol in the resveratrol biosynthetic germinated brown rice had a resveratrol content of 0.246 ug / It was confirmed that the effect of inhibiting the amount of MMP-1 protein expression was superior to that of the treatment group. The protein expression of type I procollagen increased the amount of protein I procollagen in the resveratrol biosynthetic germinated brown rice group.
As a result, it was confirmed that the resveratrol biosynthesis germinated brown rice extract effectively inhibited the expression amount of MMP-1 protein in photoaging cells induced by ultraviolet irradiation and increased the protein expression amount of type I procollagen (FIG. 3 ).
Example 5: Confirmation of wrinkle improvement effect using artificial skin
Example 5-1: Photoaging Establishment of induction artificial skin model
Human reconstructed skin, which is composed of keratinocyte and fibroblast (Keraskin TM FT) was purchased from Modern Cell & Tissue Technologies Inc. In order to measure the effect of UVA-induced photo-aging on the artificial cells, the effects of 1% (1: 1) (dissolved in PBS containing 10% propylene glycol) were measured in order to measure the effects of Dongjin germinated brown rice, resveratrol biosynthetic germinated brown rice and resveratrol. w / v) germinated brown rice, resveratrol biosynthetic germinated brown rice and 20 uL resveratrol were applied to artificial skin and irradiated with ultraviolet rays (UVB) of 100 mJ / cm 2 after 24 hours. After 24 hours, immunohistochemical staining was performed with 10% formalin fixed and hematoxylin-eosin (H & E) staining and masson's trichrome staining. The samples were incubated in PBS containing 0.1% protease for antigen retrieval and then incubated in PBS containing 3% H 2 O 2 for 10-15 minutes. The samples were then incubated in MMP-1 (Abcam, Cambridge, MA, USA) and type I procollagen (Santa Cruz Biotechnology Inc., USA) for 1 hour in PBS containing 2% normal horse serum, CA) to complete photoaging induced artificial skin model for Dongjin germinated brown rice, resveratrol biosynthesis germinated brown rice and resveratrol.
Example 5-2: Photoaging Induction artificial skin Measurement of effect on collagen layer
In order to evaluate the efficacy of the germinated brown rice, resveratrol biosynthesis germinated brown rice and resveratrol on the photoaged skin induced by ultraviolet rays, the artificial skin model prepared in Example 5-1 was subjected to hematoxylin-eosin staining and mason trichrome Histological analysis was performed using staining (Fig. 4).
The morphology of artificial cells was observed from the result of hematoxylin - eosin (HE) staining. As shown in Fig. 4, in the case of the ultraviolet (UVB) irradiation group to which the sample was not administered, collapse of the artificial skin epidermal layer was observed. A slight decay was observed in the epithelial layer in the Dongjin germinated brown rice group, but no collapse of the epithelial layer was observed in the resveratrol biosynthetic germinated brown rice group. In addition, the morphology of the collagen layer was observed from the results of mason trichrome (MT) staining. In the ultraviolet (UVB) irradiation group without the sample, collagen was decomposed and the collagen layer was found to be thinner. Collagen degradation was observed in the Dongjin germinated brown rice group, but it was confirmed that the amount and density of the collagen layer in the resveratrol biosynthesis germinated brown rice treated group were higher than those in the Dongjin germinated brown rice treated group (Fig. 4).
From these results, it was confirmed that the resveratrol biosynthesis germinated brown rice extract protects the epithelial cells from collapse of artificial cell epithelial layer due to ultraviolet irradiation, prevents collagen degradation, and prevents skin aging and skin wrinkles.
Example 5-3: Analysis of skin wrinkle-related enzyme protein expression in artificial skin
Protein expression levels of MMP-1 and type I procollagen were examined through immunohistochemical staining of photoaging induced artificial skin (Fig. 5). As shown in FIG. 5, in the expression amount of type I procollagen, when the control group to which no sample is added is irradiated with ultraviolet light (UVB +), the expression amount of type I procollagen decreases. The protein expression of type I procollagen was increased in UV irradiated-resveratrol biosynthetic germinated brown rice treated group compared with the UV irradiated-Dongjin germinated brown rice treated group. In addition, in the expression amount of MMP-1, when the control group to which the sample is not added is irradiated with ultraviolet light (UVB +), the expression amount of MMP-1 is increased (brown). The amount of MMP-1 expression was found to be decreased in the UV irradiation-resveratrol biosynthesis germinated brown rice treated group compared to the UV irradiation-Dongjin germinated brown rice treated group.
From these results, it was confirmed that resveratrol biosynthesis germinated brown rice extract increased protein expression of type I procollagen and decreased the amount of protein expression of MMP-1.
Example 6: Resveratrol Biosynthesis Germinated brown rice Long-term pay mouse skin Collagen layer analysis
The effects of resveratrol biosynthesis germinated brown rice on mouse skin tissue collagen layer were investigated. In the present invention, the amount of resveratrol used as a control group is 0.98 ug / g, and the amount of resveratrol contained in resveratrol biosynthesis germinated brown rice also contains the same amount. The samples were fed to the mice for 3 months and 6 months, and the skin tissues stained with Mason's trichrome staining were observed, and skin tissues were analyzed using Apero ScanScope FL (Leica Biosystems, Nussloch, Germany) ). In addition, the thickness of the collagen layer of the skin tissue was quantitatively analyzed using the Image-J system (NIH, Maryland, USA) program (Table 1 and FIG. 7).
As a result, as shown in Table 1 and Fig. 7, the thickness of the collagen layer of the mice fed with resveratrol biosynthetic germinated brown rice for 3 months was 327.8 ± 13.4 μm, and the collagen layer of the mouse fed with 3 g of Dongjin germinated brown rice was 203.3 ± 7.5 The thickness of the collagen layer was increased by 1.6 times or more as compared with the collagen layer 204.4 ± 10.2 μm of mice fed with um and resveratrol for 3 months. In addition, the thickness of collagen layer of mice that had been fed resveratrol biosynthesis germinated brown rice for 6 months was 444.4 ± 34.1 μm, and the collagen layer of 227.8 ± 13.4 μm of the mouse group fed with Dongjin germinated brown rice for 6 months and the collagen of the mouse fed with resveratrol for 6 months Layer, the thickness of the collagen layer was increased by 1.9 times or more and the collagen layer was increased by 2.9 times or more compared to the collagen layer of the control group without addition of the sample (150.0 ± 7.5 μm).
From these results, it was confirmed that resveratrol biosynthesized germination showed an excellent effect of suppressing skin aging and skin wrinkles by increasing the collagen layer of mice.
From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.
Claims (11)
Said composition having the features of i), ii) and iii) below;
i) inhibition of MMP-1 activity,
ii) increased activity of type I procollagen, and
iii) Increased thickness of the collagen layer.
Wherein the composition further comprises a pharmaceutically acceptable carrier.
The resveratrol biosynthetic germination
i) immersing the resveratrol biosynthetic rice seed in water; And
ii) germinating brown rice seeds immersed in the water by repeatedly germinating seeds under light and dark conditions.
The resveratrol biosynthesis germinated brown rice extract
i) grinding the resveratrol biosynthetic germinated brown rice; And
ii) adding the ground sprouted brown rice flour to a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and a mixed solvent thereof, and then extracting.
Wherein said step ii) is repeated two or more times.
Wherein the solvent is 80% methanol.
i) immersing the resveratrol biosynthetic rice seed in water;
ii) germinating rice seeds immersed in the water by repeatedly treating the rice seeds under light and dark conditions;
iii) grinding the germinated rice seeds; And
iv) adding the ground sprouted brown rice flour to a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms and a mixed solvent thereof, and extracting.
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| KR20210146647A (en) | 2020-05-27 | 2021-12-06 | 주식회사 피토메카 | Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates |
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| KR101986008B1 (en) * | 2017-06-20 | 2019-06-05 | 가천대학교 산학협력단 | Pharmaceutical composition for preventing or treating atopic dermatitis comprising extract of resveratrol-enriched rice or resveratrol enriched rice callus as an active ingredient |
| EP3777872B1 (en) * | 2018-04-10 | 2023-09-27 | Stardust HD, Inc. | Compositions comprising citrus junos seed extract and brown rice extract and uses thereof |
| KR102628991B1 (en) * | 2021-11-16 | 2024-01-25 | 고려대학교 산학협력단 | Cosmetic compositions for antioxidation, skin-whitening and anti-wrinkle comprising radicle extracts or its fractions of brown or colored rices as effective components |
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| Title |
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| Mary Ndiaye et al, ‘The grape antioxidant resveratrol for skin disorders: Promise, prospects, and challenges’, Archives of Biochemistry and Biophysics, 2011, Vol.508, 164-170* |
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| KR20210146647A (en) | 2020-05-27 | 2021-12-06 | 주식회사 피토메카 | Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates |
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