KR20200044373A - Composition for skin regeneration and wound healing containing absolute of paederia foetida flower - Google Patents

Abstract

The present invention relates to a composition for skin regeneration and a cosmetic composition, all of which contain absolute extracted from Paederia scandens flowers. The absolute provides excellent effects of skin regeneration, skin elasticity enhancement, and wound healing, through effects of cell proliferation and migration, and collagen synthesis inducement at the skin cell level.

Description

Composition for skin rejuvenation and wound treatment, including gyeyo flower absolut {COMPOSITION FOR SKIN REGENERATION AND WOUND HEALING CONTAINING ABSOLUTE OF PAEDERIA FOETIDA FLOWER}

The present invention relates to a composition for the treatment of skin regeneration and wounds containing absolutum extracted from Paederia foetida as an active ingredient, and cosmetic and skin external use thereof.

The skin is the tissue that covers the outermost part of the human body and protects the human body and functions as a barrier, thereby providing a primary defense function that protects the body from external chemical and physical shocks. The skin is composed of three layers of epidermis, dermis and subcutis, and 95% of the epidermal layer located on the outermost layer is composed of keratinocytes. Keratinocytes of the epidermis effectively defend against bacteria and chemicals penetrating the skin, and secrete various cytokines to participate in the inflammatory and immune responses of the skin. In addition, the movement and proliferation of keratinocytes not only provide wound healing and skin regeneration, but also induce collagen synthesis, which plays an important role in skin elasticity enhancement and basement membrane formation. Collagen has more than 20 kinds of collagen according to α-chain, and type I, III, IV, V, VI and VII collagen is distributed in human skin. Mainly type I collagen makes up more than 85% of the dermal layer of the skin, type IV collagen is the main structural protein of the basement membrane, and synthesis of type I and IV collagen enhances skin elasticity and improves basement membrane formation. It is known that the adhesion and proliferation of skin cells are regulated, and it improves the barrier function to selectively penetrate harmful substances and provides an interaction between the epidermis and the dermis. For this reason, the movement and proliferation of skin epidermal keratinocytes and the induction of collagen synthesis not only promotes skin regeneration and wound healing, but also promotes skin elasticity and helps maintain healthy skin.

Recently, the problems of high toxicity to humans and hormonal disturbances of various chemical synthetic substances used in cosmetics and quasi-drugs have been recognized, and it is necessary to develop natural materials. In particular, as natural fragrances are known not only for their aroma and aromatherapy effects, but also for their antibacterial and anti-inflammatory effects, research on natural fragrances derived from natural products has been highlighted. Natural fragrances are classified variously according to extract, extraction method, use and purpose, and natural fragrances derived from plants are classified into essential oil, absolute, concrete, etc. according to the extraction method. Plant-derived natural flavorings are volatile chemicals that show pharmacological effects on the human body among secondary metabolites of plants, and many terpene compounds such as alcohol, aldehyde, ketone, and ester are components. The aromatic ingredients of these natural fragrances show the effects of stress relief, fatigue recovery, muscle relaxation, etc. through the skin, circulatory system, and olfactory system, and at the same time, have therapeutic effects such as antibacterial, immunity, and anti-allergic effects. It has been used for prevention and has been used as a raw material for food flavoring and cosmetics. As consumers around the world pursue a healthy life, they became interested in products and natural ingredients using natural ingredients, and as a result, the natural perfume market is also being applied in a wide variety of industries. As such, as natural materials are required, research on the development of functional materials derived from natural products is required.

Gyeyodeung ( Paederia foetida. ) Is a perennial plant belonging to the family Rubiaceae . It grows in the southern regions of Korea and grows in sunny places at the foot of the mountain or on the seaside grass. It is 5 ~ 7 m long and has small hairs on young branches and is also known as Gyeakdeung, hollow vine because of its unique smell. A small trumpet-shaped flower blooms from July to September and is white on the outside and purple on the inside. It has been used for the treatment of inflammatory diseases using roots, stems, and fruits since ancient times. So far, research has been conducted on antibacterial activity, antioxidant activity, and antihyperlipidemia using leaves.

Korean Patent Publication No. 10-2011-0036317

In order to solve the above problems, the present invention extracts absolut from flowers such as gyeyo, and identifies its constituents, and confirms its effect on proliferation, migration, and collagen synthesis of human skin-derived keratinocyte cell line (HaCat). Elasticity enhancement, skin regeneration and wound healing efficacy were confirmed to complete the present invention.

In order to achieve the above object, the present invention provides a composition for skin regeneration and wound healing, comprising as an active ingredient an absolute extracted from a flower, Paederia foetida .

In one embodiment of the present invention, the absolute may be extracted by an organic solvent extraction method using a volatile solvent, and may be an absolute that removes insoluble wax by extracting the concrete extracted by the organic solvent extraction method with alcohol.

In one embodiment of the present invention, the absolute comprises a volatile organic compound, for example, hexanodecanoic acid (Hexadecanoic acid), linolenic acid (Linolenic acid), nonadecan (Nonadecane), Anti Ox, tetracoic acid (-) (Tetracosane (-)), Muscalure, Pentacosane, Pentadecane, 3-methyl-, Hexacosane, 10-Heneicosene, Heptacosane, Tetracosanal, Octacosane, 1-Tetracosanol, β-Tocopherol, 1-heneicosene, Vitamin E, 12-methoxyavita-8,11,13-triene-20-aluminum ( 12-Methoxyabieta-8,11,13-trien-20-al), Campesterol, Clionasterol, Cycloartenol, 24-Methylenecycloartanol and Tetra It may include one or more volatile organic compounds selected from the group consisting of Misol (Tetramisole).

In one embodiment of the present invention, the absolute may be included in the range of 1 ~ 100 μg / mL.

In one embodiment of the present invention, the absolute can induce proliferation and migration of epidermal keratinocytes. Specifically, the absolute may induce proliferation and migration of human skin-derived keratinocyte cell lines (HaCat) and collagen synthesis.

In addition, the present invention provides a cosmetic composition comprising as an active ingredient an absolute extracted from flowers such as gyeyo.

In addition, the present invention provides a composition for external application for skin comprising absolute as an active ingredient extracted from flowers such as gyeyo.

The present invention is to characterize the components of absolut extracted from flowers such as gyeyoyo, and demonstrate their skin elasticity enhancement, skin regeneration and wound healing efficacy, and induce cell proliferation and migration at the skin cell level, thereby inducing an excellent effect on skin regeneration and wound healing Have

1 is a flow chart showing the process of extracting the absolute (absolute) from the gyeyoye flowers in one embodiment of the present invention.
Figure 2 is a graph showing the toxicity to the epidermal keratinocytes of Absolut extracted from the flowers, etc. in one embodiment of the present invention.
3 is a graph showing the effect of absorptive flowers of wisteria flowers on (A) proliferation and (B) migration of epidermal keratinocytes in one embodiment of the present invention.
Figure 4 is an image showing the effect on the natural growth (outgrowth) collagen sprouts (outgrowth) in (A) epidermal keratinocytes of Absolute (A) of the wisteria flowers in one embodiment of the present invention and (B) collagen sprouts (sprouting) It is a graph that quantifies the results on outgrowth.
Figure 5 is a graph showing quantifying the effect on the (A) collagen type I synthesis, and (B) the effect on collagen type IV synthesis of Absolute of Gyeyode flowers in one embodiment of the present invention.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description, detailed descriptions of techniques well known to those skilled in the art may be omitted. In addition, in the description of the present invention, when it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, the detailed description may be omitted. In addition, terms used in the present specification (terminology) are terms used to properly express a preferred embodiment of the present invention, which may vary according to a user, an operator's intention, or customs in the field to which the present invention pertains.

Therefore, the definition of these terms should be made based on the contents throughout the specification. Throughout the specification, when a part “includes” a certain component, it means that the component may further include other components, not to exclude other components, unless otherwise stated.

The present invention relates to a composition for skin regeneration and wound healing comprising an absolute extracted from a flower such as gyeyo as an active ingredient, a cosmetic composition comprising the same, and a composition for external application for skin

The absolut is preferably obtained from flowers that are not dried, and may be absolut, which removes insoluble wax by extracting concrete extracted with an organic solvent extraction method with alcohol.

Specifically, the absolut comprises the steps of obtaining a concrete by extracting the gyoyo flowers by an organic solvent extraction method using a volatile solvent; And extracting the obtained concrete with alcohol to remove insoluble wax and obtaining an absolute.

As an example, the absolute may be extracted as shown in the flowchart of FIG. 1. First, after drying the dried flower, etc., is precipitated in hexane (n-hexane), the obtained extract is concentrated using a rotary evaporator to obtain concrete, and then ethyl alcohol is added to the obtained concrete. After storing and dissolving at -20 ° C for 12 hours, separation / purification using a vacuum cooling filtration and a rotary evaporator can be used to prepare a flower lamp absolut according to the present invention.

The organic solvent extraction method and the method of separating absolute from the extract can include any method that can be modified by a person skilled in the art.

The absolut extracted from the flowers, such as gyoyo, contains various physiologically active ingredients. More specifically, hexanodecanoic acid, linolenic acid, nonadecane, anti ox, and tetracoic acid (- ) (Tetracosane (-)), Muscalure, Pentacosane, Pentadecane, 3-methyl-, Hexacosane, 10-Heneicosene, Heptacosane, Tetracosanal, Octacosane, 1- Tetracosanol, beta-tocopherol, 1-heneicosene, vitamin E, 12-methoxyavita-8,11,13-triene-20-aluminum (12- Methoxyabieta-8,11,13-trien-20-al), campesterol, clionasterol, cycloartenol, 24-methylenecycloartanol and tetramizole ( Tetramisole) may include one or more volatile organic compounds selected from the group consisting of, but is not limited to It is not.

In addition, the content of these components is specifically 1 to 2% by weight of hexanodecanoic acid (Hexadecanoic acid), linolenic acid (Linolenic acid) 27 to 28% by weight, Nonadecane (Nonadecane) 0.1 to 1% by weight, Anti Ox 5 ~ 6% by weight, Tetracosane (-) (8) to 9% by weight, Muscalure 1 to 2% by weight, Pentacosane 2 to 3% by weight, Pentadecane, 3-methyl- 0.1 to 1% by weight , Hexacosane 4 ~ 5 wt%, 10-Heneicosene 6 ~ 7 wt%, Heptacosane 0.1 ~ 1 wt%, Tetracosanal 0.1 ~ 1 wt%, Octacosane 1 ~ 2 wt% , 1-tetracosanol (1-Tetracosanol) 5-6 wt%, beta-tocopherol (ß-Tocopherol) 4-5 wt%, 1-heneicosene 7-8 wt%, vitamin E (Vitamin E) 2-3 wt% %, 12-Methoxyabieta-8,11,13-trien-20-aluminum (12-Methoxyabieta-8,11,13-trien-20-al) 1-2% by weight, Campesterol 6 ~ 7% by weight, Clionasterol 5 ~ 6% by weight, Cycloartenol 1 ~ 2% by weight, 24-methylenecycloartano l) 0.1 to 1% by weight and tetramisole (Tetramisole) may be included within the range of 0.1 to 1% by weight.

In the present invention, it was found that the absolute extracted from gyeyo, etc. contains components exhibiting the physiological activity described above, and exhibits the effects of proliferation and migration of epidermal keratinocytes (see FIG. 3) by these components.

Accordingly, it can be used as a skin regeneration and wound healing composition that contains the absolute extracted from the flowers of the yogurt according to the present invention as an active ingredient, alone containing one or more pharmaceutically acceptable absolutes extracted from the pharmaceutical composition Carriers, excipients or diluents.

When the absolute of the present invention is used as a composition for skin regeneration and wound healing, absolute may be used at a concentration of 1 to 100 μg / mL. The effect of proliferation and migration of epidermal keratinocytes is insignificant when using Absolute Flower Absolute at a concentration of 1 μg / mL or less, and when Absolute is used at a concentration of 100 μg / mL or more, there is a marked increase in the effect of increasing content. It is not desirable because it does not appear and decreases.

In the above, "a pharmacologically effective amount" refers to an amount sufficient for the physiologically active ingredient to be administered to an animal or human to exhibit a desired physiological or pharmacological activity. However, the pharmaceutically effective amount may be appropriately changed according to the severity of symptoms, the age of the patient, weight, health status, sex, administration route, and treatment period.

In addition, "pharmaceutically acceptable" in the above refers to physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorder, dizziness or similar reaction when administered to humans. Examples of the carrier, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be further included.

In addition, the compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal, and can be formulated for oral or parenteral administration. It can be formulated in form. Formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders, ointments, lotions, creams, plasters, pastes, and the like.

The composition according to the present invention can be administered through several routes including oral, transdermal, subcutaneous, intravenous or intramuscular, and the dosage of the active ingredient is a number of factors such as the route of administration, the patient's age, gender, weight and severity of the patient. It can be appropriately selected according to. In addition, the composition of the present invention can be administered in parallel with a known compound that can enhance the desired effect.

Furthermore, the anti-obesity composition according to the present invention can be used not only as a pharmaceutical composition as described above, but also as a health functional food. For example, it can be easily utilized as a main raw material, an auxiliary raw material, a food additive, a functional food or beverage.

The "food" refers to a natural product or a processed product that contains one or more nutrients, and preferably means a state that can be directly eaten after a certain processing process, and in a general sense, food It includes food additives, functional foods and beverages.

Foods to which the food composition can be added include, for example, various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, special nutritional foods (e.g., dairy products, English, baby food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), breads, health supplements, seasoned foods (e.g. soy sauce) , Miso, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g. snacks), candy, chocolate, gum, ice cream, dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchis) , Pickles, etc.), beverages (eg, fruit drinks, vegetable drinks, soy milk, fermented drinks, etc.), natural seasonings (eg, ramen soup, etc.). The food, beverage or food additive may be prepared by a conventional manufacturing method.

In addition, the term “functional food” or “health functional food” refers to a food group or a living body having a food composition that has added value to act and express the function of the food for a specific purpose by using physical, biochemical, or biotechnical techniques. Refers to foods designed and processed to sufficiently express the body's regulation functions for defense rhythm control, disease prevention and recovery, etc. Specifically, it may be a health functional food. The functional food may include a food-acceptable food supplement additive, and may further include suitable carriers, excipients, and diluents commonly used in the production of functional foods.

The type of the dietary supplement is not limited thereto, but may be in the form of a powder, granule, tablet, capsule or beverage.

Advantages and features of the present invention, and a method of achieving them will be clarified with reference to embodiments described below in detail. Hereinafter, the present invention will be described in detail by examples. However, these examples are intended to illustrate the present invention in detail, and the scope of the present invention is not limited to these examples.

[Example]

Example 1: Preparation of gyeyoyeol flower absolute

In accordance with the process shown in Figure 1 to prepare a flower absorptive gyeyode.

In this example, the gyeyo dormitory flower was collected and used as a wild gye nymph flower native to Jeonnam Haenam. The entire flower buds just before flowering of gyeyo etc. were collected, washed, and 2 kg of fresh grass were extracted using an organic solvent extraction method.

In other words, the dried non-dry flowers were precipitated in hexane, and the precipitate thus obtained was evaporated in a rotary evaporation to concentrate to obtain solid concrete. Then, the concrete was dissolved by adding ethyl alcohol and stored at minus 20 ° C, and then insoluble wax was removed through a vacuum cooling filtration, and concentrated again using a rotary evaporator. And separating / refining to prepare a system of absorptive flowers. And the prepared gyeyodeul flower absolute was stored in a freezer at minus 80 ℃.

Example 2. Analysis of ingredients of gyeyodeung flower absolute

The analysis of the composition of the flower of Absolute of Gyoyo lamp was commissioned by KBSI (Korea Basic Science Institute, Seoul Center). A gas chromatography / mass spectrometer (6890N GC / 5975i MS, Agilent, USA) was used, and a column of GC / MS was DB5-MS (30 m × 50 μm, 0.25 μm), and carrier gas ( carrier gas) was analyzed by flowing at 1 mL / min using helium. The temperature of the oven was maintained at 40 ° C-2 ° C / min-230 ° C-5 ° C / min-300 ° C (5 minutes), and the injection temperature was analyzed at 280 ° C. In addition, the interface temperature was analyzed at 300 ℃, the ion source temperature at 230 ℃, and the analyzer temperature at 150 ℃, and the mass range was analyzed at 40 to 800 m / z.

Identification of the flower absolut component of Gyeyodeung flower was confirmed by comparing the retention time of volatile components (retention indices, RI) with that of standard materials, or by comparing the mass spectrum of volatiles with the mass spectrum of Wiley 7Nist 05 library. RI was determined using C ₆ to C ₂₄ (nalkane) under the same conditions as the sample. As a result, it was confirmed that it was composed of a total of 23 components (Table 1).

Experimental Example 1. Measurement of toxicity (cell viability) of Absolute such as gyeyo on keratinocytes

Toxicity (cell viability) on keratinocytes was investigated using the abdominal absorpt obtained in Example 1 above.

12 hours incubation the keratinocytes from 0.1 ㎎ / ㎖ of collagen (collagen, type I) into a concentration of 5 × 10 ³ / 100㎕ / well to the coated 96-well plate, and then, 37 ℃, 5% CO ₂ incubator Did. After 12 hours, abdominal light was treated with 0.1 to 200 μg / ml concentration and cultured for 24 hours. After treating EZ-cytox, absorbance was measured at 450 nm and the results are shown in FIG. 2.

As a result of the experiment, as shown in FIG. 2, the cytotoxicity of keratinocytes was not observed at the concentration of 0.1 to 200 µg / ml for abdominal light, and the cell viability increased at the concentration of 50 to 150 µg / ml. . Therefore, the concentration of the absolut component used in the present invention is preferably 200 μg / ml or less.

Experimental Example 2. Effects of flower Absolute, such as gyeyo, on the proliferation of epidermal keratinocytes

In order to confirm the effect of the abdominal flower flower absolute obtained in Example 1 on skin epidermal keratinocyte proliferation, it was investigated as follows.

Human skin-derived epidermal keratinocytes (HaCat) were cultured using DMEM medium containing 10% fetal bovine serum and utilized in experimental examples of the present invention. Epidermal keratinocytes were injected into a 96-well microtiter plate so that the number of cells per well was 3 X 10 ³ cells / 100 μL / well and attached in a 37 ° C, 5% CO ₂ incubator.

100 μL of each well was treated at a concentration of 1-100 μg / mL Absolute Flower, and incubated for 48 hours. Ez-cytox and DMEM were mixed at 1: 2, treated at 30 μL per well, and reacted for 30 minutes in an incubator. After measuring absorbance at 450 nm using an ELISA plate reader (Synerge2, BioTek, USA), it was converted to a percentage to obtain results for proliferation of cutaneous epidermal keratinocytes in flower absolut. Figure 3A is a graph showing the proliferative effect of epidermal keratinocytes of gyeyosang flower absolut extracted by Example 1 of the present invention. Here, the negative control is not using the gyeyodeul flower absolute, the positive control is using epidermal growth factor (Epithermal growth factor, EGF 50 ng / mL) that is conventionally used for wound healing and / or skin regeneration.

As a result, as shown in FIG. 3A, it has been shown that the abdominal flower absolute according to the present invention induces the proliferation of epidermal keratinocytes in a concentration-dependent manner.

That is, the proliferation of epidermal keratinocytes by epidermal growth factor (50 ng / mL) was induced to 127.20 ± 0.80%, and the positive control group compared to the 141.60 ± 8.27% proliferation induction rate at 50 μg / mL concentration of absolute flowers. It showed that the induction rate of proliferation of 200.40 ± 12.39% at a concentration of 100 μg / mL was shown to effectively induce proliferation of keratinocytes.

Experimental Example 3. Effect of Gyeyo-do Flower Absolute on Epidermal keratinocyte migration

In order to confirm the effect of the gyeyode Flower Absolute obtained in Example 1 on the skin epidermal keratinocyte migration, the experiment was conducted as follows.

Human skin-derived epidermal keratinocytes (HaCat) were cultured using DMEM medium containing 10% fetal bovine serum and utilized in experimental examples of the present invention. The mobility of epidermal keratinocytes was investigated using a 48-well Boyden microchemotaxis chamber, first treated with a concentration of 1 to 100 μg / mL, respectively, in the lower chamber at a concentration of 1 to 100 μg / mL, and polycarbonate of 8 μm pore coated with type I collagen The membrane was loaded. Subsequently, epidermal keratinocytes were treated in the upper chamber at 1 × 10 ⁶ cells / 50 μL per well. After incubating the chamber for 3 hours and 30 minutes in a 37 ° C incubator, the membrane was fixed and stained with a Diff-Quick solution.

By counting the number of cells that have passed through the membrane, and converting them into percentages, the results for the movement of the skin epidermal keratinocytes of the absolute flowers Absolute were obtained.

Figure 3B is a graph showing the effect of the movement of the skin keratinocytes of the gyeyoyeop flower absolute extracted by Example 1 of the present invention. Negative control was not used as a system of flower absolute, and positive control was using epidermal growth factor (EGP 1 ng / mL), which is conventionally used for wound healing and / or skin regeneration.

As a result, as shown in FIG. 3B, the actinus flower absolute according to Example 1 of the present invention induced concentration-dependent movement of epidermal keratinocytes, in particular, 105.96 ± 6.46 at a concentration of 10 μg / mL actinic flower blossom absolute %, And the maximum migration rate of 122.52 ± 5.87% at a concentration of 50 μg / mL, and it was found that the flower absolute of indigo plant effectively induces the movement of keratinocytes.

Experimental Example 4. The effect of gyeyo-de-flower Absolute on epidermal keratinocyte collagen sprouting out growth

In order to measure the effect of the syllup flower absolute obtained in Example 1 on the synthesis of epidermal keratinocyte collagen type I and collagen type IV, the following investigation was conducted.

After mixing Type I collagen and epidermal keratinocytes to 2.5 × 10 ⁷ cells / mL, neutralize by treating with NaOH (1 N) solution, spotting 5 μL in a 24 well plate and drying at room temperature for 10 minutes. Artificial skin tissue was formed. Samples were processed by concentration and incubated in a 37 ° C., 5% CO ₂ incubator for 48 hours. The culture solution was removed, and cells and tissues were fixed and stained using Diff-Quik solution, and the stained cells and tissues were photographed to obtain results.

Figure 4 is a result showing the effect on the out growth of epidermal keratinocytes collagen sprouting of the backbone flower absolute extracted by Example 1 of the present invention, the negative control is not using the backbone flower absolute, positive control is conventional It is using epidermal growth factor (EGF 1 ng / mL) that is used for wound healing and / or skin regeneration.

As a result, as shown in Fig. 4A, it has been shown that the absolute flower absolut induces concentration-dependent outgrowth of epidermal keratinocyte collagen sprouting. That is, it was found that in comparison with the positive control, it was found to effectively induce outgrowth of keratinocyte collagen sprouting at a concentration of 50 μg / mL flower absorptive. In particular, as shown in FIG. 4B, 749.43 ± 15.56% was induced at a concentration of 50 μg / mL abdominal flower, and 586.78 ± 50.23% was induced at a concentration of 100 μg / mL.

Experimental Example 5. Effects of Gyeyo-do Flower Absolute on the synthesis of epidermal keratinocyte collagen type I and collagen type Ⅳ

In order to measure the effect on the outgrowth of collagen sprouting of epidermal keratinocytes, the system of the flowers of abseil obtained by Example 1 was investigated as follows.

A monoclonal anti-collagen I or IV Ab (2 μg / well / PBS) was injected into a 96 well microtiter plate and coated at room temperature for 12 hours. After blocking for 1 hour at room temperature with a blocking buffer, the sample was treated with 100 μL per well and reacted at room temperature for 90 minutes. The sample was treated with a culture medium (CM) of epidermal keratinocytes that had not been treated with abdominal flower absorption and a culture medium (CM) of epidermal keratinocytes treated with 10 mg / mL concentration of absolute flower absolut, 100 mg / mL concentration of absolute flower absolute A culture medium (CM) of the epidermal keratinocytes was treated. Thereafter, 100 μL of each polyclonal anti-collagen I or IV Ab was treated and reacted at room temperature for 90 minutes, and then avidin-HRP was treated at each well for 60 minutes at room temperature. After injecting 100 μL of each supersignal ELISA pico chemiluminescent substrate for 10 minutes and reacting for 10 minutes, the degree of luminescence was measured using an ELISA plate reader. The results for were secured as shown in FIG. 5. In particular, as shown in FIG. 5A, synthesis of collagen type I at a concentration of 100 μg / mL in the flower of the back light was induced by 140.01 ± 8.96%, as shown in FIG. 5B, of collagen type IV at the concentration of 100 μg / mL in the flower of the back light Synthesis was 153.43 ± 9.60%.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Claims (8)

Gyeyodeung ( Paederia foetida. ) A composition for skin regeneration and wound healing that includes absolute extracted from flowers as an active ingredient. According to claim 1,
The absolute is a composition for skin regeneration and wound treatment that is extracted by an organic solvent extraction method using a volatile solvent. According to claim 2,
The absolute is a composition for skin regeneration and wound healing that is an absolute that removes insoluble wax by extracting the concrete extracted by the organic solvent extraction method with alcohol. According to claim 1,
The absolute is hexanodecanoic acid (Hexadecanoic acid), linolenic acid (Linolenic acid), nonadecane (Nonadecane), Anti Ox, tetracoic acid (-) (Tetracosane (-)), Muscalure, Pentacosane, Pentadecane, 3-methyl-, Hexacosane, 10-Heneicosene, Heptacosane, Tetracosanal, Octacosane, 1-Tetracosanol, Beta-Tocopherol, 1-heneicosene, vitamin E, 12-methoxyavita-8,11,13-triene-20-aluminum (12-Methoxyabieta-8,11,13-trien-20-al), campesterol ( Skin comprising one or more volatile organic compounds selected from the group consisting of Campesterol, Clionasterol, Cycloartenol, 24-Methylenecycloartanol and Tetramisole Composition for regenerative and wound healing. The method according to any one of claims 1 to 4,
The absolute is a composition for skin regeneration and wound treatment that is included in the range of 1 to 100 μg / mL. The method according to any one of claims 1 to 4,
The absolute is a composition for skin regeneration and wound treatment that induces proliferation and migration of epidermal keratinocytes. Gyeyo lamp ( Paederia foetida. ) A cosmetic composition comprising as an absolute active ingredient extracted from flowers. Gyeyodeung ( Paederia foetida. ) A composition for external application for skin comprising absolute extracted from flowers as an active ingredient.

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