KR101902911B1 - Composition for improving human skin cell injury by UV-B comprising the extract of Hydrangea serrata - Google Patents

Abstract

The present invention relates to a composition for improving human epidermal cell and dermal cell injury caused by ultraviolet rays, which contains an extract of mountain syrup as an active ingredient.
According to the present invention, the composition of the present invention not only restores UV-B-induced cell damage in human epidermal cells and dermis cells, but also increases the secretion of hyaluronic acid and inhibits the secretion of MMP-1, May indicate improved cell damage. Therefore, the safflower extract as a safe material can be usefully used in the fields of food composition or cosmetics.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for improving human skin cell damage by ultraviolet rays,

More particularly, the present invention relates to a composition for improving human epidermal and dermal cell damage caused by UV-B exposure of skin containing an extract of Scr. .

Skin aging can be classified into intrinsic aging and extrinsic aging depending on factors. Endogenous aging is caused by changes in physiological functions of skin epidermis and dermis according to age. Exogenous aging is known to be caused by changes in physiological functions of skin caused by environmental pollution, exposure to ultraviolet rays, and stress. Oxidative stress induced by UV among these mechanisms of aging increases free radicals in the body and increases the activity of MMP-1 which degrades collagen and hyaluronic acid Decomposition causes skin epidermal and dermal damage by increased activity of hyaluronidase.

In everyday life, we are always exposed to sunlight, especially cell damage and skin cancer caused by UV-B exposure is an inevitable factor in everyday life.

Therefore, researches on the inhibition of skin cell damage by UV exposure have been actively carried out, and researches on cosmetics, health functional foods and medicines showing biochemical efficacy and efficacy have been actively conducted. The main research subjects are materials with practical effects and effects due to the development of research level and the increase of technology.

Plant-derived materials have been used for a long time because of their excellent safety, and in particular, functional materials based on plant and herbal ingredients used in the private sector or used in oriental medicine have been actively developed in the country.

Hydrangea serrata is a broad-leaved tall tree of the submandibular gland, and its leaf parts are indicated as edible parts in food raw materials notified in the place of shipment. It is used as a treatment for chronic bronchitis, warts, genomes, and anti-inflammatory drugs. Korean Patent Laid-Open Publication No. 2004-0063874 discloses a composition containing an extract of Saccharomyces cerevisiae having hyperglycemia and hyperlipidemia inhibitory activity.

However, the direct mechanism of inhibition of human epidermal and dermal cell damages by ultraviolet rays of mountainous plants has not been studied yet. Therefore, the inventors of the present invention have conducted studies on the antioxidant enzyme activity by ultraviolet rays of the extracts and the direct basic effects on the inhibition of human epidermal cell and dermal cell damages.

Thus, the present inventors have earnestly studied in order to overcome the problems of the prior art sought results, arithmetic station ROS removed through human skin cells and antioxidant activity in dermal cells containing an extract of (Hydrangea serrata) as an active ingredient and UV- B of the present invention can exert a skin improving effect by eliminating skin damage, and the present invention has been completed.

KR 10-2004-0063874 A KR 10-2005-0056569 A

 WOO, M. S., et al. Comparison of skin elasticity test results from the Ballistometer® and Cutometer®Skin Research and Technology, 2014, 20.4: 422-428.  IMOKAWA, Genji. Mechanism of UVB-induced wrinkling of the skin: paracrine cytokine linkage between keratinocytes and fibroblasts leading to the stimulation of elastase. In: Journal of Investigative Dermatology Symposium Proceedings. Nature Publishing Group, 2009. p. 36-43.

Accordingly, it is a main object of the present invention to provide a composition for improving human epidermal cell and dermal cell injury by ultraviolet rays, which comprises an extract of Scr.

According to one aspect of the present invention, there is provided a composition for preventing and ameliorating damage to human skin cells by ultraviolet rays containing an extract of hydrangea serrata as an active ingredient.

The present invention provides a composition for preventing and / or ameliorating human skin cell damage by ultraviolet light, wherein the human skin cell damage by ultraviolet rays is human epidermal cell and dermal cell damage caused by UV-B exposure. The experimental example of the present invention not only restores the cell damage caused by UV-B in human epidermal cells and dermis cells, but also increases the secretion of hyaluronic acid and suppresses the secretion of MMP-1, .

In the present invention, " damage " of human skin cells includes death of human skin cells due to ultraviolet rays, damage of skin cell DNA, increase of reactive oxygen species, increase of lipid peroxidation, and the symptoms include erythema, Deposition, photoaging, skin cancer, and the like. In addition, " prevention " of damage refers to any action that inhibits or delays skin cell damage by the ultraviolet light. In addition, " improvement " of damage refers to any action that alleviates damage to the skin cells caused by the ultraviolet rays or reduces the degree of symptoms.

In the present invention, the extract can be extracted with any solvent conventionally used for extracting natural plants, and is preferably extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof . The alcohol may be at least one selected from the group consisting of ethanol, methanol, isopropanol, and butanol, preferably ethanol, more preferably a fermented alcohol.

In the present invention, the extract may be contained in an amount of 0.001 to 80.0% by weight, preferably 5 to 20% by weight, based on the total weight of the composition. In addition, it was confirmed that the above conditions show a high yield and excellent recovery from UV-B.

In one embodiment of the present invention, the UV light may be UV-B.

According to the experimental examples of the present invention, it can be seen that ethanol extract of mountain water plant can improve the damage caused by UV-B exposure of the skin (see Experimental Examples 1 and 2).

In the present invention, the composition has at least one form selected from the group consisting of tablets, granules, pills, capsules, liquids, jellies or gums, and is characterized in that it is for oral consumption.

In the present invention, the composition has at least one formulation selected from the group consisting of a softening agent, a nutritional lotion, a nutritional cream, a water cream, a spot, a gel lotion or an ointment.

The composition of the present invention comprising the extract of Scutellaria baicalensis as an active ingredient can be used for various purposes, for example, as a cosmetic composition, a pharmaceutical composition, a health functional food composition and the like.

The cosmetic composition according to 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 oil, water, a surfactant, a moisturizer, a lower alcohol, , A chelating agent, a coloring agent, a preservative, a perfume, and the like may be appropriately compounded, but the present invention is not limited thereto.

The pharmaceutical composition or health functional food composition according to the present invention may further comprise an appropriate carrier, excipient, diluent or the like conventionally used in the production of a pharmaceutical composition. The pharmaceutically acceptable carrier, excipient or diluent is selected from the group consisting of lactose, dextrose, sucrose, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, Benzoate, talc, magnesium stearate, and mineral oil. However, the present invention is not limited thereto.

The pharmaceutically effective amount and the effective dose of the pharmaceutical composition according to the present invention may vary depending on the formulation method, administration method, administration time and / or administration route of the pharmaceutical composition. The type and degree of the reaction to be achieved by the administration of the pharmaceutical composition, the kind of the subject to be administered, age, weight, general health condition, symptom or degree of disease, sex, diet, excretion, The components of the drug or other composition used in conjunction therewith, and the like, and similar factors well known in the medical arts. One of ordinary skill in the art can readily determine and prescribe dosages effective for the desired treatment. Administration of the pharmaceutical composition according to the present invention can be administered once a day, or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention. A preferred dose of the pharmaceutical composition according to the present invention may be from 1 mg / kg to 1, OOO mg / kg per day.

As described above, the composition containing the extract of the present invention of the present invention promotes the activity of SOD (superoxide dismutase), inhibits ROS caused by UV in skin cells, It is suitable as a composition for skin care by restoring the damage. In addition, the composition containing the extract of the present invention of the present invention increases secretion of hyaluronic acid and inhibits secretion of MMP-1, thereby preventing skin aging caused by ultraviolet rays and maintaining skin elasticity. This is a safe material and can be usefully used in food composition field or cosmetic field.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Preparation of an extract using ethanol

The extract of Scutellariae Radix contained in the composition of the present invention is prepared by the following procedure. Firstly, a 1 year cultivating station, which is a native species in Jeju Island, was purchased, washed thoroughly with distilled water, and completely dried in a cool, sunny place until there was no change in weight. Thereafter, the completely dried dried material was pulverized by using a homogenizer to obtain pulverized products, and 400 g of an extraction solvent was prepared by mixing ethanol: distilled water at a weight ratio of 7: 3, and then 50 g of the pulverized product was added. At this time, the mixing ratio of the pulverized material to the extraction solvent was 1: 8 by weight. Next, the extraction solvent mixed with the pulverized product was put into a shaking constant-temperature water bath and stirred at 50 DEG C for 2 hours. The material obtained by extraction was insoluble matter removed at room temperature using Wattman (No. 2) extraction solvent filter paper. Thereafter, the solvent was completely removed while concentrating under reduced pressure in a distillation apparatus equipped with a cooling condenser. After extracting the extracts, the extraction yield was 20%.

Experimental Example 1: Evaluation of cytotoxicity of ethanol extract of mountain water plant

The extract obtained in Example 1 was assayed for cytotoxicity against skin cells. In this experiment, HaCaT keratinocyte was used as keratinocyte and HS68 fibroblast was used as dermal cell. To confirm cytotoxicity, each cell of 1.0 × 10 ⁴ cells was placed on a 96-well plate and allowed to adhere for 24 hours to stabilize the cells at a concentration of 0, 5, 25, 125 ppm (μg / ml) The ethanol extract of mountain water plant was treated. After 24 hours, cell viability was measured by MTT assay. MTT assay is a method for measuring the amount of formazan formed from MTT reduced by mitochondria of living cells. Each cell was treated with 50 mg of 5 mg / ml MTT solution and incubated for 4 hours. The solution was completely removed and the plate dissolved in DMSO was measured at 540 nm absorbance.

extract Cytotoxicity of extract (%) 0 μg / ml 5 μg / ml 25 μg / ml 125 μg / ml Epidermal cell 100.0 108.0 100.3 113.9 Dermal cell 100.0 104.7 102.9 95.6

As shown in the above Table 1, the extract of Scutellaria japonica was treated by concentration and cytotoxicity test was conducted. As a result, it was confirmed that there was no cytotoxicity in dermal and epidermal cells.

EXPERIMENTAL EXAMPLE 2 Evaluation of Cell Damage Recovery by UV-B of Ethanol Extract of Mountain Water

The extracts obtained in Example 1 were measured for damage to skin cells exposed to UV-B. In this experiment, HaCaT keratinocyte was used as keratinocyte and HS68 fibroblast was used as dermal cell. To confirm cell repair efficacy, 1.0 x 10 ⁴ cells of each cell were dispensed on 96-well plates, stuck for 24 h, stabilized and irradiated with UV-B at 15 mJ / cm ² . Subsequently, the cells were treated with ethanolic extracts of Oryzias latipes at concentrations of 0, 5, 25 and 125 ppm (μg / ml) and cell viability was measured by MTT assay. MTT assay is a method for measuring the amount of formazan formed from MTT reduced by mitochondria of living cells. More specifically, 50 쨉 l of 5 mg / ml MTT solution was added to each of the cells and incubated for 4 hours. The solution was completely removed and the plate dissolved in DMSO was measured at 540 nm absorbance.

 extract UV-induced recovery of skin cell damage (%) 0 μg / ml 0 μg / ml 5 μg / ml 25 μg / ml 125 μg / ml UV-B
15 mJ / cm ² - + + + + Epidermal cell 100 80.46 96.45 100.10 110.20 P-value - 0.000 * 0.000 * 0.000 * Dermal cell 100 76.48 104.1 100.53 167.7 P-value - 0.020 * 0.130 0.010 *

P-value: Statistical treatment between group (0 μg / ml) treated with t-test and group treated with extract

As shown in the above Table 2, when the UV-B treatment at 15 mJ / cm ² was performed, the cell damage was 80% and 76% in the epidermal and dermal cells, respectively. As a result of UV-induced skin cell injury repair, it was found that cell damage was restored statistically significantly (P-value <0.05). Considering that the composition of the present invention is an extract, a relatively high recovery effect can be confirmed in a concentration-dependent manner.

Experimental Example 3: Analysis of hyaluronic acid content

The extract obtained in Example 1 was assayed for increasing hyaluronic acid in dermal HS68 fibroblast. To confirm that the hyaluronic acid could be increased by the treatment of the material, 1.0 × 10 ⁵ cells of each of the cells were dispensed on a 24-well plate and stuck for 24 h and then stabilized at 0, 5, 25, 125 ppm (μg / ml) and cultured for 24 hours in a medium supplemented with ethanol extract of mountain water. The degree of secretion of hyaluronic acid was measured using a TECO ^ⓡ Hyaluronic Acid PLUS ELISA Kit (TE 1018-2, TECO Medical Group, US) in the supernatant treated with the substance.

extract Hyaluronic Acid (ng / ml) 0 μg / ml 5 μg / ml 25 μg / ml 125 μg / ml Dermal cell 88.0 83.0 124.6 119.6 P-value 0.300 0.000 * 0.000 *

P-value: Statistical treatment between group (0 μg / ml) treated with t-test and group treated with extract

As shown in Table 4, the hyaluronic acid test was conducted by treating the extracts of Scutellaria japonica with the concentrations of 25% and 125 μg / ml in the dermal cells, and statistically significant (P-value <0.05) Secretion promoting effect was confirmed.

Experimental Example 4: Analysis of MMP-1 content

For the extract obtained in Example 1, it was determined whether MMP-1 could be increased in the epidermal cell HaCaT Keratinocyte. To determine if MMP-1 could be reduced by treatment with the material, 1.0 × 10 ⁵ cells were seeded on 24-well plates for 24 h and stabilized by attachment at 0, 5, 25, 125 ppm μg / ml), and cultured for 24 hours. After that, the secretion level of MMP-1 was measured in the supernatant treated with MMP-1 Human ELISA Kit (ab100603, abcam, US).

extract MMP-1 (pg / mL) 0 μg / ml 5 μg / ml 25 μg / ml 125 μg / ml Epidermal cell 10613 9703 9813 8263 P-value 0.010 * 0.030 * 0.000 *

P-value: Statistical treatment between group (0 μg / ml) treated with t-test and group treated with extract

As shown in the above Table 5, MMP-1 treatment was carried out by treating the extract of Araliae sativum L. with various concentrations, and the inhibitory effect of MMP-1 on the epidermal cells was confirmed statistically significantly (P-value <0.05).

Formulation Example 1: Preparation of tablets

The extracts obtained in Example 1 were mixed with the components shown in Table 5 according to a conventional tablet preparation method and tableted to prepare tablets.

Raw material name Unit weight (mg) The extract of Example 1 50 Corn starch 100 Lactose 100 Stearic acid 2

Formulation Example 2: Preparation of capsules

The ingredients obtained in Example 1 were mixed with the ingredients shown in Table 6 according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Raw material name Unit weight (mg) The extract of Example 1 50 Corn starch 100 Lactose 100 Stearic acid 2

Formulation Example 3: Preparation of liquid agent

The ingredients obtained in Example 1 were mixed with the ingredients listed in Table 7 according to the preferred beverage preparation method, and filled into bottles or pouches to prepare a liquid preparation.

Raw material name Unit weight (g) The extract of Example 1 2.5050 Xanthan gum 0.0075 Fructooligosaccharide solution 0.7500 Coconut flower essence powder 1.0500 Twin condensate 1.5000 Red ginseng 0.0450 Purified water 9.1425

Formulation Example 4: Preparation of jelly

The extracts obtained in Example 1 were mixed with the ingredients shown in Table 8 according to the preferred jelly preparation method, and the jellies were prepared by filling three kinds of jelly.

Raw material name Unit weight (g) The extract of Example 1 2.0000 Food gel 0.3600 Carrageenan 0.0600 Calcium lactate 0.1000 Sodium citrate 0.0600 Composite Golden Extract 0.0200 Enzyme treatment Stevia 0.0440 Fructooligosaccharide solution 5.0000 Red grape concentrate 2.4000 Purified water 13.9560

Formulation Example 5: Preparation of nutritional cream

Nutritive creams were prepared for the extracts obtained in Example 1 according to the usual methods according to the composition shown in Table 9 below.

Raw material content The extract of Example 1 1.0 Sitosterol 4.0 Polyglyceryl 2-olate 3.0 3.0 Setares-4 2.0 cholesterol 3.0 Dicetyl phosphate 0.4 Concentrated glycerin 5.0 Sunflower oil 22.0 Carboxyvinyl polymer 0.5 Triethanolamine 0.5 antiseptic a very small amount Spices a very small amount Purified water Balance

The above composition ratio is generally prepared by mixing suitable components to formulate the formulation. However, the blending ratio and the raw materials may be arbitrarily changed as necessary.

The extract of the present invention was stable in all formulation test conditions and therefore had no problem in the stability of the formulation.

Claims (6)

A composition for preventing and ameliorating damage to human skin cells by ultraviolet rays containing an extract of Hydrangea serrata as an active ingredient.
The composition according to claim 1, wherein the human skin cell damage caused by ultraviolet rays is human epidermal cell or dermal cell damage caused by UV-B exposure.
The composition according to claim 1, wherein the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
The composition for preventing and treating damage to human skin cells according to claim 1, wherein the extract comprises 0.001 to 80% by weight based on the total weight of the composition.
The composition according to claim 1, wherein the composition has at least one formulation selected from the group consisting of tablets, granules, pills, capsules, liquids, jellies or gums, and is for oral ingestion. A composition for preventing and improving damage.
The composition according to claim 1, wherein the composition has at least one formulation selected from the group consisting of softening agent, nutritional lotion, nutritional cream, water cream, spot, gel lotion or ointment, A composition for preventing and improving skin cell damage.