KR20220090815A - Composition for preventing or improving skin cell damage caused by ultraviolet rays containing Symplocos chinensis f. pilosa (Nakai) Ohwi fruits extracts - Google Patents

Abstract

The present invention relates to a composition for preventing or improving skin cell damage caused by ultraviolet rays or an antioxidant composition comprising the Norinjae tree extract, and the present invention relates to a Norinjae tree extract, which can effectively restore skin cells damaged by ultraviolet rays, and effective the same It is possible to provide a pharmaceutical composition, cosmetic composition or food composition comprising as a component.

Description

Composition for preventing or improving skin cell damage caused by ultraviolet rays containing Symplocos chinensis f. pilosa (Nakai) Ohwi fruits extracts}

The present invention relates to a composition for preventing or improving skin cell damage caused by ultraviolet rays, and more particularly, to a composition for preventing or improving skin cell damage caused by ultraviolet rays, comprising an extract from the fruit of Norinjae tree.

Skin aging can be divided into intrinsic aging and extrinsic aging according to factors. It is known that intrinsic aging is caused by changes in the physiological functions of the skin epidermis and dermis according to age, and extrinsic aging is caused by changes in the physiological functions of the skin caused by environmental factors such as air pollution, UV exposure, and stress. Among these mechanisms of aging, oxidative stress induced by UV increases free radicals in the body, increases the activity of MMP-1 that decomposes collagen, and reduces hyaluronic acid. Decomposition causes damage to the epidermis and dermis of the skin due to increased activity of hyaluronidase.

In our daily life, we are always exposed to sunlight, and in particular, cell damage and skin cancer caused by UV-B exposure are unavoidable factors in our daily life.

As a result, studies on the mechanism of suppression of skin cell damage caused by UV exposure have been actively conducted, and studies on cosmetics, health functional foods, and pharmaceuticals showing biochemical efficacy and effects are being actively conducted. Materials with practical efficacy and effects are the main research subjects due to the development of the research level and the increase in technology.

Symplocos chinensis f. pilosa (Nakai) Ohwi is a Symplocos chinensis f. pilosa (Nakai) Ohwi, distributed in China, Japan, and India. The branches are hairy. Leaves are alternate, ovate or long oval-ovate, with sharp ends, usually with small saw teeth on the edge, but sometimes indistinctly and alternately. The fruit is 8mm long, oval, and ripens in a wall color in September. The panicle is 4-8cm long, hangs at the tip of a new branch, has hairs on the peduncle, and the petals are long oval and open to the side. Flowers are 8-10mm in diameter and bloom white in May. The petals are long oval, with many stamens, and can be enjoyed for about a fortnight and have a subtle fragrance.

It is known for the treatment of diabetes, diabetic complications, and nervous system analgesia, but the effect of preventing or ameliorating the skin cell damage caused by UV rays is not known. It was intended to be developed as a material for

Unexamined Patent Publication No. 10-2020-0086207 Registered Patent Publication No. 10-1132040

In order to solve the above problems, the present invention is to provide an extract of Norinjae tree effective for preventing or improving skin cell damage.

In addition, it is intended to provide a composition for preventing or improving skin cell damage comprising the Norinjae tree extract.

In addition, it is intended to provide an antioxidant composition comprising the Norinjae tree extract.

In order to achieve the above object, the present invention provides a Norinjae tree extract.

In addition, it provides a composition for preventing or improving skin cell damage comprising the Norinjae tree extract.

In addition, there is provided an antioxidant composition comprising the Norinjae tree extract.

In the present invention, there is provided a composition for preventing and improving human skin cell damage due to ultraviolet light, characterized in that the damage to human skin cells by ultraviolet rays is damage to human epidermal cells and dermal cells caused by exposure to ultraviolet rays.

Specifically, it not only repairs cellular damage caused by UV rays in human epidermal cells and dermal cells, but also reduces the amount of MMP-1, MMP-2, MMP-3, MMP-9 or pro-collagen (procollagen) that is reduced by UV rays. The expression level can be restored to the level of the normal group.

In the present invention, "damage" of human skin cells includes apoptosis of human skin cells by ultraviolet rays, DNA damage of skin cells, increase in reactive oxygen species, increase in lipid peroxidation, and the like, and the symptoms include erythema, sunburn, pigmentation. deposition, photoaging, skin cancer, and the like. In addition, “prevention” of damage refers to any action that inhibits or delays the damage to skin cells by the ultraviolet rays. In addition, "improvement" of damage refers to any action of alleviating the damage state of the skin cells caused by the ultraviolet rays or reducing the degree of symptoms.

The ultraviolet rays include, but are not limited to, UV-A and UV-B.

The antioxidant means the action of inhibiting oxidation, and although prooxidant and antioxidant are in balance in the human body, this balance is unbalanced by various factors and leads to oxidation promotion. When tilted, oxidative stress is induced, leading to potential cell damage and pathological diseases. Reactive oxygen species (ROS), the direct cause of oxidative stress, are unstable and highly reactive, so they easily react with various biomaterials and attack the body's polymers to cause irreversible damage to cells and tissues, mutations, or mutations. It causes cytotoxicity and carcinogenesis.

Reactive nitrogen species (RNS) such as NO, HNO2, and ONOO are produced in large amounts due to the immune response of macrophages, neutrophils and other immune cells during the inflammatory response, and ROS is also generated at this time. The active oxygen as described above oxidizes and destroys cells in the body, and thus is exposed to various diseases. In addition, the antioxidant refers to the function of inhibiting oxidation of cells by highly reactive free radicals or reactive oxygen species (ROS) depending on intracellular metabolism or oxidative stress caused by the influence of ultraviolet rays. and removing free radicals or reactive oxygen species, thereby reducing cell damage.

In addition, the antioxidant effect in the present invention includes the ability to scavenging free radicals such as ABTS and DPPH, and the ability to inhibit or remove active oxygen such as ORAC and ROS.

The Norinjae extract is obtained by extracting the whole plant, or a part of the plant (root, leaf, flower, stem fruit, etc.), such as the above-ground part of the plant (leaf, flower, stem, fruit, etc.), preferably the stem, leaf, It may be obtained by extracting the fruit, and more preferably may be a Norinjae fruit extract, but is not limited thereto.

The Norinjae tree extract may be extracted with any solvent conventionally used to extract natural plants, and preferably may be extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. As the alcohol, any one or more selected from the group consisting of ethanol, methanol, isopropanol, and butanol may be used, and ethanol may be preferably used.

The ethanol is not limited, but 30 to 90% (v/v) ethanol, preferably 60 to 80% (v/v), more preferably 70% (v/v) ethanol may be used.

The composition may be any one selected from a food composition, a cosmetic composition, and a pharmaceutical composition.

When the Norinjae tree extract is included in a food composition, a cosmetic composition, or a pharmaceutical composition, the concentration is not limited, but preferably more than 0 to 500 µg/ml, more preferably 0.1 to 300 µg/ml, more preferably 0.1 to 100 μg/m may be included. When it exceeds the above range, the prevention or improvement effect of skin damage is not significantly improved.

The pharmaceutical composition may be prepared in a form generally prepared when preparing a pharmaceutical composition in the art, and may be prepared in the form of tablets, granules, pills, capsules, liquids, and the like. In addition, the pharmaceutical composition may further include additives, excipients, carriers, etc. commonly used in the art for preparing a pharmaceutical composition.

The cosmetic composition may be prepared in a form that is generally prepared when manufacturing a cosmetic composition in the art, and examples include lotion, cream, lotion, and the like. In addition, the cosmetic composition may further include additives, emulsifiers, surfactants, humectants, preservatives, and the like, which are conventionally added when preparing a cosmetic composition in the art.

The food composition may be prepared in a form generally prepared when preparing a food composition in the art, and may further include additives, pigments, fragrances, etc. that are commonly added when preparing a cosmetic composition in the art .

The present invention relates to a composition for preventing or improving skin cell damage caused by ultraviolet rays and an antioxidant composition comprising the Norinjae tree extract, and the present invention relates to a Norinjae tree extract that can effectively restore skin cells damaged by ultraviolet rays and effective the same A pharmaceutical composition, cosmetic composition or food composition comprising as a component may be provided.

1 shows the results of MMP-1, -3, -9 expression and type-1 procollagen mRNA expression according to UV-B irradiation in skin cells treated with Norinjae tree extract according to the present invention.

Hereinafter, the present invention will be described in detail by way of Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

<Example> Preparation of Norinjae tree extract

As an extract of Hemniferae tree fruit, mature Hemniferae fruit was collected in the fall. The collected fruits were immediately pulverized, dried in the shade at room temperature, or dried in a dryer at 50 ° C. To increase the extraction yield, dried seeds were pulverized with a grinder, and the extraction solvent was prepared by extraction using 70% (v/v) ethanol.

<Experimental Example 1> Intracellular Collagen by UV-B treatment, Matrix metalloprotease (MMP-1, -2, -3, -9), Tissue inhibitor of Metalloproteinases (TIMP-1, -2), Reversion-inducing-cysteine -Activity assay for skin anti-aging factors such as rich protein (RECK)

In order to confirm the prevention or improvement effect of the Norinjae tree extract according to the present invention on the prevention or improvement of skin cell damage caused by UV rays, UV-B in the skin cells was irradiated at 30mJ/cm ² to reduce TIMP-1, -2, RECK and Collagen An experiment was conducted to determine whether the reduction and MMP activity were effectively inhibited.

The experimental group treated with 0, 4, 20, and 100 μg/ml of Norinjae tree extract of Example was irradiated with UV-B at 30 mJ/cm ² to confirm the expression levels of MMP-1, -2, -3, -9, etc. . As a result, it was confirmed that the expression levels of MMP-1, -2, -3, and -9, which were reduced by UV-B, increased significantly, and the decreased expression level of pro-collagen was also significantly increased ( FIG. 1 ).

<Experimental Example 2> Antioxidative activity evaluation of Norinjae tree extract according to the present invention

In order to evaluate the antioxidant capacity of the Norinjae tree extract according to the present invention, the antioxidant activity was evaluated as follows.

2-1. ABTS, DPPH, ORAC assay

ABTS assay: ABTS cation (ABTS ⁺ ) was formed after mixing 7mM ABTS and 2.45 mM potassium persulfate and incubating at room temperature for 16 hours. Thereafter, the test solution was prepared by diluting so that the absorbance value at 734 nm was 1.5 or less. 10 μl of the sample was added to 1 ml of the previously prepared ABTS ⁺ solution, and the absorbance value was measured after 5 minutes.

DPPH assay: 0.2 ml of sample was added to 0.8 ml of 0.4 mM DPPH solution dissolved in 95% ethanol, mixed, and then incubated for 5 minutes in the dark, and absorbance was measured at 517 nm.

ORAC assay: First, 2,2'-azobis (2-methylpropionamidine) dihychloride (AAPH) was used to generate peroxy radicals, and trolox was used as a standard material. The sample was prepared by dissolving it in 75 mM phosphate buffer, and then incubated at 37° C. for 15 minutes. After that, 150 μl of 0.2M fluorescein was added to 25 μl of the sample, and 25 μl of 0.15 nM AAPH was immediately added. After that, measurements were made at a wavelength of 485nm-530nm for 60 minutes (once every 2 minutes). The concentration of trolox used as a standard was measured as 0, 0.625, 1.25, 2.5, 5, 10, and 20 μM, and the inhibitory ability of the sample was expressed as trolox equivalents.

2-2. Measurement of inhibitory effect on excessive production of active oxygen (ROS) in skin cells by UV irradiation

Intracellular ROS generated by UV irradiation on skin cells was measured using DCFH-DA staining dye. DCFH-DA is a type of fluorescent dye that rapidly enters the cell, and when reactive oxygen species in the cell increase, DCFH-DA is converted to DCF-DA and shows a green fluorescence value. After culturing the cells, UV was irradiated, and the medium was removed after further culturing for a certain period of time. Then, DCFH-DA at a concentration of 4 μM was dissolved in HBS and the cells were treated. After incubation at 37° C. for 30 minutes, it was washed three times with PBS. Thereafter, the change in fluorescence value was measured using the FL-1 wavelength of FACs.

2-3. Measurement of skin cell death inhibitory effect by UV irradiation

Apoptosis caused by UV irradiation on skin cells was measured using MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) reagent. MTT reagent measured the degree of apoptosis by dissolving the formazan formed to form formazan by dehydrogenase of living mitochondria. After culturing the cells, the medium was removed after irradiating UV light and culturing for a certain period of time. Thereafter, MTT at a final concentration of 0.5 mg/mL was dissolved in PBS and treated. After incubation at 37 ° C. for 4 hours, the supernatant was removed and the formazan formed by treatment with 100 uL of 100% DMSO was dissolved. Thereafter, the absorbance was measured at 540 nm using an ELISA device to measure the degree of apoptosis, and the results are shown in Table 1.

Antioxidant activity and UV protection effect of Norinjae tree fruit extract black activity ORAC assay
IC50 (mg/ml) 1.2 ABTS assay
IC50 (mg/ml) 7.3 DPPH assay
IC50 (mg/ml) 4.0 Protective effect of UVB-treated cell death
EC50 (μg/ml) 24.7 Inhibitory effect of UVB-treated
ROS generation
IC50 (μg/ml) >250

Claims (7)

A composition for preventing, improving, or antioxidizing skin cell damage caused by ultraviolet rays, comprising an extract of Norinjae tree. According to claim 1, wherein the Norinjae tree extract is skin cell damage prevention, improvement or antioxidant composition, characterized in that extracted from the fruit Norinjae tree. According to claim 1, wherein the Norinjae tree extract is water, 60 to 80% (v / v) ethanol, or a mixture thereof for preventing, improving or antioxidant composition for skin cell damage caused by ultraviolet rays, characterized in that the extraction . According to claim 1, wherein the Norinjae tree extract is contained in a concentration of more than 0 to 500μg / ml for preventing, improving or antioxidant composition for skin cell damage caused by ultraviolet rays. [Claim 3] The composition for preventing, improving, or antioxidizing skin cell damage caused by UV rays according to claim 1, wherein the skin cell damage caused by UV rays is caused by UV exposure. The method according to claim 1, wherein the Norinjae tree extract increases the expression level of MMP-1, MMP-2, MMP-3, MMP-9 or procollagen. or a composition for antioxidants. According to claim 1, wherein the composition is a pharmaceutical composition, a cosmetic composition, characterized in that any one selected from the food composition for preventing, improving or antioxidant composition for skin cell damage caused by ultraviolet rays.

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