KR20210002045A - Composition comprising extract of Dictyota coriacea - Google Patents

Abstract

The present invention relates to a composition comprising an extract of Dictyota coriacea as an active ingredient. The Dictyota coriacea extract according to the present invention suppresses collagenase activity, suppresses elastase activity to increase skin regeneration or anti-wrinkle effects, reduces the total amount of melanin to exhibit a skin whitening effect, and inhibits the production of nitric oxide involved in the inflammatory reaction to exhibit an anti-inflammatory effect. Thus, the Dictyota coriacea extract can be used in pharmaceuticals, cosmetics, or health foods.

Description

Composition comprising extract of Dictyota coriacea {Composition comprising extract of Dictyota coriacea}

The present invention relates to a composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, comprising an extract of bamboo networm extract.

Skin is the frontmost defense organ of the human body that protects our body from external stimuli, and its interest is increasing from an aesthetic point of view. Accordingly, research on various functional cosmetics is active, and among them, various functional cosmetics for wrinkle relief are being released.

Skin aging is largely divided into two types, internal aging and external aging, depending on the cause. Intrinsic aging is an aging process that is naturally caused by deteriorating physiological functions of the body as age increases. And, external aging (extrinsic aging) is an aging phenomenon caused by external factors such as ultraviolet (UV), dry air, reactive oxygen species and stress. As skin aging progresses due to these factors, wrinkles are created on the skin surface.

Various causes of wrinkles have been reported. First, when the skin is exposed to excessive ultraviolet rays, collagen, a component of the skin, is accelerated to decompose, resulting in loss of elasticity and wrinkles. In addition, when the skin becomes excessively dry due to excessive temperature change and humidity reduction or wind, etc., the skin function as a barrier against the outside decreases, resulting in wrinkles. In addition, when the skin is exposed to reactive oxygen species or free radicals, lipid peroxidation is generated by an oxidation action, and thereby, proteins constituting the skin such as collagen are deformed, resulting in wrinkles.

On the other hand, when skin aging occurs, keratinization of the skin occurs, the fusion rate of epidermal cells gradually slows, the epidermal cell layer becomes thinner, the stratum corneum layer becomes thick, and the boundary line with the dermal layer becomes a straight line. In the dermis, due to the aging of human skin fibroblasts, the productivity of fiber and matrix decreases, the level of peroxide in the cell increases rapidly, and it is affected by various contaminants and stress, making it impossible to perform the function of cells smoothly. The metabolic activity of cells decreases, and as a result, wrinkles are formed due to functional and structural damage, thereby promoting skin aging. Therefore, various cosmetic compositions are being studied in order to suppress the occurrence of skin aging or wrinkles caused by these external and internal factors.

On the other hand, Dictyota coriacea (Holmes) refers to brown algae belonging to Phaeophyceae, Dictyotales, and Dictyotaceae. It is a brown, thin dengue shape, splits into two branches several times, becomes a branch, spreads widely in a fan shape, and grows, the roots are short string-shaped, the branches are slightly serpentine or twisted once or twice, and the armpits of the branches are narrow, 20~ It is a marine plant that grows up to 30 cm, and the cross section of the branch is made of cells with two layers of chloroplasts, and the water tissue is made of one layer of large cells, and grows wild in the south coast or off the coast of Jeju Island.

It is an object of the present invention to provide a composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including extracts of bamboo nets of a bamboo net.

The present invention provides a pharmaceutical composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including extracts of bamboo nets, skewers.

In addition, the present invention provides a cosmetic composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including the extract of bamboo nets for participation.

In addition, the present invention provides a health functional food for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including the extracts of bamboo nets for participating in the present invention.

The extract according to the present invention exhibits a whitening effect by inhibiting collagenase activity, inhibiting elastase activity, improving skin regeneration or wrinkles, reducing the total amount of melanin, and being involved in inflammatory reactions. It can be used in medicines, cosmetics, or health foods by inhibiting the production of nitric oxide and showing anti-inflammatory effects.

1 is a schematic diagram showing the separation and purification development of 80% ethanol extract of participating bamboo nets (Dictyota coriacea).
FIG. 2 is a graph showing the result of separating the Sephadex LH-20 column active fraction of the extract of the participating bamboo nets, using HPLC.
Figure 3 is a graph showing the results of measuring the molecular weight of the HPLC active fraction of the extract of the participant bamboo netmulbatang using a Q-TOF mass.
Figure 4 is a graph showing the results of measuring the collagenase inhibitory activity IC50 of the Sephadex LH-20 column fractions of the extract of the extract of bamboo networm.
5 is a graph showing the results of measuring cell viability for fibroblasts CCD-986SK of fractions F1-T-F17.
6 is a graph showing the results of collagenase inhibitory activity using Western blot of the F1-T-F17 fraction.
7 is a graph showing the results of Melanin contents assay in an artificial cell model (Neoderm-ME).
8 is a photograph and graph showing the results of measuring the Melanin content (Fontana-Masson stain) in an artificial cell model (Neoderm-ME).
9 is a photograph showing the result of confirming the anti-Ki-67 antibody inhibitory activity using Immunohistochemistry (IHC) stain of the F1-T-F17 fraction.
10 is a graph showing the results of the inhibitory activity of Collagenase type IV antibodies using Immunohistochemistry (IHC) stain of the F1-T-F17 fraction.
11 is a graph showing the results of the anti-inflammatory activity (NO assay) of the F1-T-F17 fraction.
12 is a photograph showing the results of anti-inflammatory activity (morphological analysis) of fractions F1-T-F17.
13 is a photograph showing the results of anti-inflammatory activity (Western blot) of the F1-T-F17 fraction.

The present invention provides a composition comprising the extracts of bamboo nets, a participant as an active ingredient.

In the preparation of the extract of the present invention, dried or undried, dried or undried, or a mixture thereof may be used. For the efficiency of extraction, you can crush and use the participant porridge. In addition, in order to obtain an effective skin regeneration, wrinkle improvement, whitening or anti-inflammatory extracts, before crushing the participating bamboo nets, blanching is performed at a high temperature of about 100°C for about 5 minutes. It can go through the process of inactivating the enzyme in the cell.

In the present invention, for the preparation of the extract of bamboo nets, it can be extracted according to a conventional extraction method using an extraction solvent of 3 to 10 times that of the leaves of bamboo nets.

As the extraction solvent, water, an organic solvent, or a mixed solvent thereof widely used in extraction of natural products may be used. The organic solvent includes ethanol, methanol, hexane, chloroform, and ethalacetate. Such an organic solvent may be mixed with water, but may be used without mixing with water for easy elution of the active ingredient. In one embodiment, the participant bamboo net base extract may be an ethanol extract.

The extract can be further fractionated using an organic solvent having a polarity different from that of the organic solvent used in the first extraction. In this case, the participant bamboo net base extract may be a participant bamboo net base extract fraction obtained by extracting the participant bamboo net base with an organic solvent and fractionating the residue with an organic solvent having a different polarity.

In addition, Sephadex LH-20, silica gel column chromatography, thin layer chromatography (TLC), and high-performance liquid chromatography for the participant bamboo grass extract or participant bamboo grass extract fraction Further purified fractions may be obtained by purification using various chromatography such as (high performance liquid chromatography, HPLC). In the present invention, the'chamskin net base extract' includes all extracts, fractions and purified products obtained in each step of extraction, fractionation, or purification, and their dilutions, concentrates or dried products.

In one embodiment of the present invention, the extract may be a hexane fraction obtained by extracting the parting bamboo net base with ethanol, and fractionating the residue with hexane and ethyl acetate.

Sephadex LH-20, column chromatography may be repeatedly performed on the obtained hexane fraction, or TLC and HPLC may be performed to obtain a further purified extract fraction.

In the following examples, fractions exhibiting skin regeneration, wrinkle improvement, whitening and anti-inflammatory activity were prepared from the extract through this additional purification process.

The present invention also provides a method for preparing a fraction of the extract of the extract of the extract of the extract of the extract of bamboo grass, comprising the step of obtaining a hexane fraction by extracting the extract of the bamboo grass with ethanol, and fractionating the residue with a mixed solvent of hexane and ethyl acetate.

In an example of the present invention, a residue obtained by extracting a participant bamboo net base with 80% ethanol was fractionated with hexane:ethyl acetate to prepare a participated bamboo net base extract fraction. In addition, the hexane fraction of the extract fraction of the participating juknetmulbatangmal was further fractionated and purified. Participation bamboo net base extract of the present invention obtained through this process and the extract fraction of participating bamboo net base extract exhibit excellent skin regeneration, wrinkle improvement, whitening and anti-inflammatory effects.

Therefore, the participant bamboo net base extract and the participant bamboo net base extract fraction of the present invention may be included as active ingredients for skin regeneration, wrinkle improvement, whitening and anti-inflammatory.

In the present invention, the term'skin regeneration effect' refers to recovery of skin tissue against damage caused by external and internal causes of the skin. The damage caused by the external cause may include ultraviolet rays, external pollutants, wounds or trauma, and the damage caused by the internal cause may include stress.

In the present invention, the term "wrinkle improvement effect" refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating wrinkles that have already been generated.

In the present invention, the term'whitening effect' not only brightens the skin tone by inhibiting the synthesis or promoting decomposition of melanin pigment, but also prevents skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones, or genetics. It says to improve.

In the present invention, "anti-inflammatory effect" refers to suppressing inflammation. Inflammation is one of the defense responses of biological tissues to certain stimuli, and refers to a complex lesion that causes tissue deterioration, circulatory disorders, effusion, and tissue proliferation. More specifically, inflammation is part of innate immunity, and, as in other animals, innate immunity in humans recognizes patterns on the cell surface that are specific to pathogens. Phagocytes recognize cells with such surfaces as non-magnetic and attack pathogens. If pathogens break through the body's physical barriers, an inflammatory reaction occurs. The inflammatory reaction is a non-specific defense action that creates a hostile environment for microorganisms invading the wound. In the inflammatory reaction, when a wound or an external infectious agent enters the body, white blood cells responsible for the early stage immune response gather and express cytokines. Therefore, the amount of expression of cytokines in cells becomes an indicator of activation of the inflammatory response. Examples of skin diseases related to inflammation include atopic dermatitis, psoriasis, erythematous disease caused by radiation, chemical substances, or burns; Itching due to acid burns, bullous skin disease, lichen planus disease or allergies; Inflammatory hair loss such as seborrheic eczema, rose acne, pemphigus vulgaris, polymorphic exudative erythema, nodular erythema, balanitis, vulvitis or alopecia areata; Cutaneous T-cell lymphoma and the like, but are not limited thereto.

The composition of the present invention inhibits collagenase activity, inhibits elastase activity to increase skin regeneration or wrinkle improvement effect, decreases the total amount of melanin to exhibit a whitening effect, and inhibits the production of nitric oxide involved in the inflammatory reaction. It can be used in medicines, cosmetics or health foods by showing anti-inflammatory effects.

The composition may be used with not only the active ingredient of the present invention, but also known skin regeneration, wrinkle improvement, whitening and anti-inflammatory components.

The pharmaceutical composition containing the extract of the extract of the present invention can be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes an excipient, a disintegrant, Sweetening agents, binders, coating agents, expanding agents, lubricants, lubricants or flavoring agents may be used.

The content of the participant bamboo networm extract in the pharmaceutical composition can be appropriately adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, the method of administration and the form of the formulation, etc., for example, 0.0001 To 10 parts by weight or 0.001 to 1 parts by weight may be included, but is not limited thereto. The content ratio is a value based on the amount of dry solvent removed.

For administration, the pharmaceutical composition can be preferably formulated as a pharmaceutical composition by including at least one pharmaceutically acceptable carrier in addition to the above-described active ingredient.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets. Further, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The composition of the present invention can be provided in the form of an external preparation for skin for skin regeneration, wrinkle improvement, anti-inflammatory or whitening.

When used as an external skin preparation, additionally fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic emulsifiers , Nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic activators, lipophilic activators, or lipid vesicles, etc. Any commonly used in skin external preparations It may contain adjuvants commonly used in the field of dermatology, such as other ingredients of. In addition, the above ingredients may be introduced in amounts generally used in the field of dermatology.

In addition, when provided as an external preparation for skin, it may be a formulation such as an ointment, patch, gel, cream, or spray, but is not limited thereto.

In addition, the present invention provides a cosmetic composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including the extract of the participating bamboo nets.

In the case of using the composition of the present invention as a cosmetic, the cosmetic product prepared by containing the extinct bamboo net base extract as an active ingredient may be prepared in the form of a general emulsified or solubilized formulation. For example, a lotion such as a flexible lotion or a nutritional lotion; Emulsions such as facial lotion or body lotion; Creams such as nourishing cream, moisture cream, or eye cream; essence; Cosmetic ointment; spray; Gel; pack; Sunscreen; Makeup base; Foundations such as liquid type, solid type or spray type; powder; Makeup removers such as cleansing cream, cleansing lotion, and cleansing oil; It may have a formulation such as a cleaning agent such as a cleansing foam, soap, and body wash.

In addition, the cosmetics include fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Water, ionic emulsifiers, nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic activators, lipophilic activators or lipid vesicles, etc. It may contain adjuvants commonly used.

In the case of a wash-off type cosmetic such as a makeup remover or detergent, in which the active ingredient stays on the skin within a short period of time when it is commercialized as a cosmetic product, it may contain a relatively high concentration of participatory bamboo networm extract. . On the other hand, in the case of leave-on-type cosmetics such as lotions, emulsions, creams, or essences in which the active ingredient stays on the skin for a long period of time, the participating bamboo net powder extract has a lower concentration than the wash-off type cosmetics. It would be okay to include. Although not limited thereto, in one embodiment of the present invention, the composition comprises 0.0001% by weight to 10% by weight (preferably 0.0001% by weight to 1% by weight) based on the total weight of the composition. can do. If the composition of the present invention contains less than 0.0001% by weight of the extract of the extract of bamboo networm, sufficient skin regeneration, wrinkle improvement, anti-inflammatory or whitening effect cannot be expected, and if it contains more than 10% by weight, allergy, etc. This is to prevent unwanted reactions or skin safety problems.

In addition, the present invention provides a food composition for skin regeneration, wrinkle improvement, whitening or anti-inflammatory, including the extract of the participating bamboo nets.

In the present invention, the term'food composition' is a food prepared by adding or encapsulating, powdering, suspension, etc. to food materials such as beverages, teas, spices, chewing gum, confectionery, etc. If it does, it means that it brings a specific effect on health, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time using food as a raw material.

The food composition according to the present invention can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages , Tea, drinks, alcoholic beverages and vitamin complexes, and includes all foods in the usual sense.

In the case of using the extract of the extract of bamboo nets, the extract can be used as a food additive, or may be used with other foods or ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be added within a range that does not impair the original taste of the food, and is usually 0.01 to 50 parts by weight or 0.1 to 20 parts by weight based on 100 parts by weight of the total composition with respect to the target food. In addition, in the case of food in the form of granules, tablets or capsules, it is usually added in the range of 0.1 to 100 parts by weight, or 0.5 to 80 parts by weight. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

The food composition may further contain various flavoring agents or natural carbohydrates. The natural carbohydrates described above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, or sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin or aspartame may be used. In addition to the above, the food composition according to the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohols, carbonates used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients may be used independently or in combination. Although the proportion of these additives is not very important, it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the food composition of the present invention.

The present invention will become apparent with reference to the following embodiments in detail the advantages and features thereof, and a method of achieving them. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, only the present embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the technical field to which the present invention pertains. It is provided to completely inform the scope of the invention to those who have it, and is only defined by the scope of the claims of the invention.

[Example]

Example 1: Preparation of Participated Bamboo Net Batangmar Extract

Participating bamboo nets (Dictyota coriacea) were washed with water, completely dried at 40℃, ground, and used as an extraction raw material.

Participated bamboo net powder was extracted according to the method shown in FIG. Specifically, 3L of 80% ethanol was added to 1 kg of powdered participant porridge, followed by stirring in dark conditions for 24 hours extraction, followed by filtration and concentration to obtain an ethanol extract. At this time, the extraction yield of the ethanol extract of the participating bamboo net base was 11.26%.

The concentrated ethanol extract was dissolved in 50 ml of a hexane:ethyl acetate solution, and then extracted through a liquid/liquid extraction method. Then, the hexane layer and the ethyl acetate layer were separated. The fractionated hexane layer and ethyl acetate layer were concentrated under reduced pressure to determine a certain amount of weight, and then the test of the experimental example to be described later was performed.

The extraction yield of the hexane fraction of the participating bamboo nets was 5.18%.

Example 2: Sephadex LH-20 and separation and purification using TLC method

Separation and purification of the hexane layer (hexane fraction), which is the active fraction separated in Example 1, was performed as shown in the schematic diagram of FIG. 1.

First, the hexane layer was separated in Sephadex LH-20 and separated into 4 fractions (F1 to F4).

Thereafter, the F1 fraction was separated and purified using TLC. TLC separation can fractionate organic solvent extracts into non-polar and polar substances, and a total of 18 fractions were separated using an eluent prepared with hexane, ethyl acetate and acetone.

F1-T-F17 fractions of the fractions were purified using HPLC.

2 is a graph showing the results of separating the Sephadex LH-20 column active fraction of 80% ethanol extract of participating juknetmulbatangmal using HPLC.

In addition, Figure 3 is a graph showing the molecular weight measurement results of the HPLC active fraction using a Q-TOF mass. The active material can be separated through the above process.

Experimental Example 1. Measurement of Total Polyphenol Content of Participating Bamboo Net Buttercup Extract

The total polyphenol content of the 80% ethanol extract and its solvent fractions isolated in the Examples are shown in Table 1 below.

Samples TPC (mg/g) EtOH ex 14.33 n-Hexane fr 74.47 EtOAc fr 33.80

As shown in Table 1, it can be seen that the total polyphenol content of the n-Hexane fraction of the 80% ethanol extract of the participating bamboo nets was excellent.

Experimental Example 2. Evaluation of skin regeneration or anti-wrinkle effect of the extract of bamboo net snail extract

1) Collagenase inhibition and elastase inhibition activity evaluation

Collagenase inhibitory activity was measured by the following method.

Human fibroblasts were placed in a 96 well plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal calf serum at 5,000 cells/well, and cultured until 90% proliferation. Thereafter, the cells were cultured in a serum-free DMEM medium for 24 hours, and then the extracts of Examples were treated in a serum-free DMEM medium, and then a cell culture solution was collected. Thereafter, the obtained cell culture solution was measured for collagenase production using a commercially available collagenase measuring instrument.

In addition, the elastase inhibitory activity was measured by the following method.

Human fibroblasts were placed in a 96 well plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal calf serum at 5,000 cells/well, and cultured until 90% proliferation. Thereafter, the cells were cultured in a serum-free medium for 24 hours, and the extracts of Examples were treated in a serum-free medium, and then a cell culture solution was collected.

Thereafter, the obtained cell culture solution was measured for the degree of elastase production using a commercially available elastase measuring instrument.

Table 2 below shows the results of the evaluation of collagenase inhibition and elastase inhibition activity of the ethanol extract, hexane fraction, and ethyl acetate fraction of the participating bamboo nets prepared in Example 1.

IC ₅₀ Value (mg/mL) Collagenase inhibitory assay Elastase inhibitory assay EtOH ex 0.15 1.06 n-Hexane fr 0.13 0.47 EtOAc fr 0.14 0.76

As shown in Table 2, it can be seen that the hexane fraction has the most excellent inhibitory activity of collagenase and elastase.

In addition, FIG. 4 is a graph showing the results of measuring the collagenase inhibitory activity IC ₅₀ of the Sephadex LH-20 column fractions (F1 to F4) prepared in Example 2.

As shown in FIG. 4, it can be seen that F1 has the most excellent collagenase inhibitory activity.

2) Cell viability measurement evaluation for fibroblast CCD-986SK

5 is a graph showing the results of measuring cell viability for fibroblasts CCD-986SK of fractions F1-T-F17.

As shown in FIG. 5, it can be seen that the F1-T-F17 fraction exhibits a high cell viability for CCD-986SK, and the cell viability increases as the content of the fraction increases.

3) Western blot measurement evaluation

Western blot evaluated the expression of MMP-1, TIMP-1 and beta-actin proteins according to the concentration of F1-T-F17 in UV-stimulated CCD-986SK fibroblasts.

6 is a graph showing the results of collagenase inhibitory activity using Western blot of the F1-T-F17 fraction.

As shown in FIG. 6, the F1-T-F17 fraction exhibits excellent collagenase inhibitory activity, and it can be seen that the activity increases as the content of the fraction increases.

4) Evaluation of anti-Ki-67 antibody inhibitory activity

Anti-Ki-67 antibody inhibitory activity was evaluated for anti-wrinkle activity (anti-Ki-67 antibody) by F1-T-F17 in Neoderm-ED using IHC staining.

9 is a photograph showing the result of confirming the anti-Ki-67 antibody inhibitory activity using Immunohistochemistry (IHC) stain for the F1-T-F17 fraction. Specifically, (A) shows a case of treatment with PBS, (B) L-ascorbic acid, and (C) F1-T-F17.

As shown in FIG. 9, it can be seen that the fraction F1-T-F17 has excellent anti-wrinkle activity.

5) Collagenase IV antibody inhibitory activity evaluation

Collagenase IV antibody inhibitory activity was evaluated for anti-wrinkle activity (collagenase IV antibody) by F1-T-F17 in Neoderm-ED using IHC staining.

10 is a graph showing the results of collagenase IV antibody inhibitory activity using IHC stain of the F1-T-F17 fraction. Specifically, (A) shows a case of treatment with PBS, (B) L-ascorbic acid, and (C) F1-T-F17.

As shown in FIG. 10, it can be seen that the fraction F1-T-F17 has excellent anti-wrinkle activity.

Experimental Example 3. Evaluation of the whitening effect of the extract of the participating bamboo nets

The melanin content was measured in melanoma cells B16F10 (Melanin contents assay).

Specifically, the target cells were inoculated into a 12-well plate and cultured for 24 hours. Subsequently, the medium was replaced, and each sample was treated with the extract of the example with 1 μM of α-melanocyte stimulating hormone (α-MSH) and cultured for 48 hours. And after washing with phosphate buffered saline (Phosphate Buffered Saline: PBS), trypsin (Trypsin)-EDTA was treated to recover the cells. The recovered cells were washed with PBS, treated with 100 μl of 1N NaOH containing 10% DMSO, heated at 80° C. for 1 hour, and cooled. And the melanin content was confirmed by measuring the absorbance at 405 nm using an ELISA microplate reader.

When measuring the melanin content, PBS was used as a control, and L-ascorbic acid was used as a positive control.

7 is a graph showing the results of melanin contents assay in an artificial cell model (Neoderm-ME).

As shown in FIG. 7, it can be seen that F1-T-F17 according to the present invention has superior melanin inhibitory activity than L-ascorbic acid, which is a positive control.

In addition, Figure 8 is a photograph and graph showing the melanin content measurement (Fontana-Masson stain) results in the artificial cell model (Neoderm-ME). In FIG. 8, (A) shows a photograph of PBS, (B) L-ascorbic acid, (C) F1-T-F17, and (D) melanin content calculated using the Image J program. Represents.

As shown in FIG. 8, it can be seen that F1-T-F17 according to the present invention has very excellent melanin inhibitory activity compared to L-ascorbic acid, which is a positive control.

Experimental Example 4. Evaluation of Anti-inflammatory Effect of Participated Juk Netting Horse Extract

The anti-inflammatory effect was evaluated as follows.

Raw 264.7 cells were used to evaluate the NO production inhibitory effect of the extracts of the examples. First, the sample was pretreated for 30 minutes, treated with 1 mg/ml LPS, incubated for 24 hours, and then the NO production inhibitory ability was evaluated using a NO quantification kit as follows.

NO production inhibitory ability (%) = {1-(NO production amount when sample is added/NO production amount when no sample is added)} x 100

11 is a graph showing the results of the anti-inflammatory activity (NO assay) of the F1-T-F17 fraction, and shows the inhibition of nitric oxide production by F1-T-F17 in LPS-stimulated Raw 264.7 macrophages.

As shown in FIG. 11, it can be seen that the F1-T-F17 fraction has an excellent anti-inflammatory effect.

In addition, Fig. 12 is a photograph showing the results of anti-inflammatory activity (morphological analysis) of fractions F1-T-F17. Specifically, it shows the morphological change of Raw 264.7 macrophages visualized by an optical microscope (x400). Cells were pretreated with F1-T-F17 before incubation with LPS for 72 hours, (A) LPS(-), (B) LPS(+), and (C) to (F) F1- T-F17 shows a case where 10 μg/mL, 25 μg/mL, 50 μg/mL, and 100 μg/mL were treated, respectively.

As shown in FIG. 12, it can be seen that the treatment of the F1-T-F17 fraction has excellent nitric oxide inhibitory activity.

In addition, FIG. 13 is a photograph showing the results of the anti-inflammatory activity (Western blot) of the F1-T-F17 fraction, showing the results of treatment with 50 μg/mL of Raw 264.7 macrophages and co-treatment with LPS for 72 hours.

As shown in FIG. 13, it can be seen that F1-T-F17 inhibits iNOS and COX-2 in Raw 264.7 macrophages.

Claims (15)

A pharmaceutical composition for skin regeneration or wrinkle improvement, comprising an extract of bamboo nets, as an active ingredient.
The method of claim 1,
Participating bamboo net base extract is a pharmaceutical composition for skin regeneration or wrinkle improvement that is an ethanol extract of participating bamboo net base horse.
The method of claim 1,
A pharmaceutical composition for skin regeneration or wrinkle improvement, which is a hexane fraction obtained by extracting the participating bamboo nets with ethanol, and fractionating the residue with hexane and ethyl acetate.
A cosmetic composition for skin regeneration or wrinkle improvement, comprising the extract of bamboo nets, as an active ingredient.
A health functional food for skin regeneration or wrinkle improvement that contains extracts of bamboo nets, as an active ingredient.
A pharmaceutical composition for whitening comprising the extracts of bamboo nets, as an active ingredient.
The method of claim 1,
Participated bamboo net base extract is a pharmaceutical composition for whitening that is an ethanol extract of participating bamboo net base.
The method of claim 1,
A whitening pharmaceutical composition, which is a hexane fraction obtained by extracting the extract of bamboo networm with ethanol, and fractionating the residue with hexane and ethylacetate.
A cosmetic composition for whitening comprising an extract of bamboo nets, as an active ingredient.
A health functional food for whitening that contains extracts of bamboo nets, as an active ingredient.
A pharmaceutical composition for anti-inflammatory comprising the extracts of bamboo nets, as an active ingredient.
The method of claim 1,
Participated bamboo net base extract is an anti-inflammatory pharmaceutical composition that is an ethanol extract of participating bamboo net base.
The method of claim 1,
Participated bamboo net base extract is a pharmaceutical composition for anti-inflammatory that is a hexane fraction obtained by extracting participant bamboo net base with ethanol and fractionating the residue with hexane and ethyl acetate.
A cosmetic composition for anti-inflammatory containing extracts of bamboo nets, as an active ingredient.
A health functional food for anti-inflammatory containing extracts of bamboo nets, as an active ingredient.

Images