KR20210155987A - Composition Comprising Complex Extract of seaweeds Fermented by Lactobacillus Brevis with Anti-Wrinkling and Elasticity, Antioxidant, and Skin Hydration Property as Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for skin elasticity or skin wrinkle alleviation, antioxidant, and skin moisturization, comprising a fermented seaweed complex extract which is fermented with a Lactobacillus brevis strain as an active ingredient. The composition of the present invention has no cytotoxicity, has an excellent effect on skin elasticity or skin wrinkle alleviation, and has an excellent effect on the antioxidant and skin moisturization, thereby being usefully used for skin elasticity or skin wrinkle alleviation in cosmetics and foods.

Description

Composition Comprising Complex Extract of seaweeds Fermented by Lactobacillus Brevis with Anti-Wrinkling and Elasticity, Antioxidant, and Skin Hydration Property as Active Ingredient}

The present invention relates to a composition for skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing, and more specifically, skin wrinkle improvement comprising a complex fermented extract of Japanese apricot fermented with Lactobacillus brevis strain as an active ingredient, It relates to a composition for antioxidant and skin moisturizing.

Some of the substances that play an important role in the process for maintaining homeostasis of an organism are physiologically active substances derived from various living organisms. Until now, many studies have been conducted on numerous physiologically active substances, and among them, research on substances having physiological activity isolated from microorganisms, plants, or animals, etc. is a very important field in life science and medicine.

On the other hand, collagen is present in the dermal layer of the skin, and is known as a major component that gives elasticity to the skin together with elastin, which accounts for about 70 to 80% of the total dry weight of the skin. Collagen is reduced by internal factors such as a decrease in cell activity due to natural aging, and biosynthesis is reduced or decomposition is promoted by external factors such as increased stress in various harmful environments or an increase in reactive oxygen species caused by sunlight. have.

On the other hand, overproduction of reactive oxygen species (ROS) has been reported in a UVB exposure environment, and the signaling systems associated with UVB exposure include mitogen activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK), It is related to proteins such as p38 kinase and c-Jun amino-terminal kinase (JNK), and induces increased expression of NF-ĸB and AP-1 transcription factors. In addition, AP-1 protein is associated with the activity of matrix metalloproteinase (MMP) gene, and NF-ĸB protein affects the expression of proinflammatory cytokines such as interleukin.

The breakdown of collagen and elastin in aged skin is mainly caused by increased expression of its degrading enzymes, such as the MMP protein. In addition, proinflammatory cytokines interfere with the synthesis of collagen and promote the breakdown of collagen. Type I collagen is the most abundant protein in the extracellular matrix of connective tissue. The extracellular matrix includes other types of proteins such as type III, V, and VII collagen, elastin, proteoglycans, and fibronectin.

Controlling the expression level of type I collagen protein is the most important factor in preventing skin photoaging. UVB irradiation reduces the expression of procollagen, a collagen precursor, in skin fibroblasts through inhibition of the TGF-β/Smad signaling system. In addition, the TGF-β1 signaling system is one of the important factors regulating the synthesis of extracellular matrix in human skin fibroblasts.

Therefore, the production of reactive oxygen species (ROS) regulated by AP-1 and MAPKs factors, and the expression of MMPs, ILs, TGF-β, and type I procollagen act as useful markers in skin photoaging studies.

As such, there have been various attempts to search for natural substances that are effective in improving skin elasticity and wrinkles by inhibiting collagen reduction. In order to take advantage of the skin elasticity and wrinkle improvement effects of collagen, products that contain collagen in external skin compositions such as cosmetics or ointments have been released. It was difficult to show the intrinsic wrinkle improvement effect. In order to solve this problem, interest in collagen synthesis promoting materials has increased, and conventionally known collagen synthesis promoting materials include vitamin C, retinoic acid, transforming growth factor (TGF), and animal placenta-derived protein (JP8). -231370), betulinic acid (JP8-208424), chlorella extract (JP9-40523, JP10-36283, fibroblast proliferation promoting action), and the like. However, there is a problem in that the use of the material is limited due to safety problems such as irritation and redness when applied to the skin, or the effect is insignificant, so that the effect of improving skin elasticity and wrinkles cannot be expected.

On the other hand, a cosmetic composition for moisturizing generally used for the skin serves to maintain a certain amount of moisture in human hair or skin, thereby making it soft and lively, and preventing damage such as cracking and dryness. That is, the cosmetic composition for moisturizing the skin is used for the purpose of beautifying the skin or hair by supplying or retaining moisture to the skin or hair in a predetermined amount or more, and keeping them healthy. The skin is an organ that has an essential function to maintain moisture, and it is known that it accounts for about 2/3 of the weight in moisture control. Therefore, in the cosmetic field, functional research related to supply and maintenance of moisture, that is, moisturizing, is being actively conducted. In addition, in recent years, the development of cosmetics that increase the moisturizing power of the skin by introducing physiologically active substances obtained from natural products into cosmetics to further strengthen the inherent defense function of the skin itself is an active trend.

Therefore, there is an urgent need for the development of natural materials that are safe for the living body, have stable active ingredients, and are superior to conventional substances known to have skin wrinkle improvement, antioxidant and skin moisturizing effects above all else.

Under this background, the present inventors conducted a study on a substance having an excellent skin condition improvement effect, and paid particular attention to the skin condition improvement effect of domestic native marine plants, improving skin elasticity or wrinkles, antioxidant, and skin moisturizing effect As a result of diligent research efforts to further improve the skin condition, the present invention was completed by identifying the excellent skin condition improvement effect of the complex fermented extract of Japanese apricot, kelp, and motherwort fermented with the Lactobacillus brevis strain.

Therefore, the main object of the present invention is to improve skin elasticity or skin wrinkles, comprising a complex fermented extract of Japanese daisy, kelp, and mother-of-pearl fermented with Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain as an active ingredient; An object of the present invention is to provide a composition for antioxidant and skin moisturizing.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention is Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain fermented with a complex fermented extract of shiitake, kelp, and mother-of-pearl as an active ingredient Skin elasticity or skin wrinkles A cosmetic composition for improving, antioxidant and skin moisturizing is provided.

The present inventors searched for the effects of various natural substances in order to find substances with excellent skin elasticity or wrinkle improvement, antioxidant, and moisturizing effects, and further improved their efficacy by paying attention to the moisturizing and wrinkle-improving effects of marine plant materials As a result of research efforts, it was found that the complex extracts of shiitake, kelp, and mother-of-pearl, and the complex fermented extract fermented with Lactobacillus brevis strains, have very excellent effects in the areas of skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing. , it was confirmed that there is no problem in safety even when applied to the skin (see FIG. 1 ).

"Hizikia fusiforme " of the present invention means a seaweed of the brown algae mosque family. Hibiscus is a perennial seaweed that lives in the lower intertidal zone and exists only in the meteorous generation. In addition, the "kelp ( Saccharina japonica )" refers to seaweeds of the kelp family of the kelp family. In addition to various pigments such as carotene, xanthophylls, and chlorophyll, kelp contains a lot of carbohydrates such as mannite and laminarin, which are produced by carbon assimilation, and alginic acid, a component of the cell wall, and amino acids such as iodine, vitamin B2, and glutamic acid. contains this In addition, the " sargassum fulvellum" refers to seaweeds of the brown algae plant, algae algae.

The term "extract" used in the present invention refers to an extract obtained according to the extraction treatment of a plant, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, such as an extract It includes extracts of all formulations that can be formed by themselves and using extracts. The extract of the present invention may be prepared and used in the form of a dry powder after extraction.

The extracts of Japanese kelp, kelp, and motherwort of the present invention may be extracted from natural, hybrid, or variegated seaweeds, and may also be extracted from seaweed tissue cultures.

As used herein, the term 'fermented extract' refers to a culture result obtained from a culture obtained by inoculating and culturing a Lactobacillus brevis strain in a medium in which a Lactobacillus brevis strain can grow. In the present invention, it refers to a culture supernatant obtained from a culture obtained by inoculating a Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain into a complex extract of Hachibana, kelp, and mother-of-pearl, or a dried product of the culture supernatant.

In the cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidation and skin moisturizing of the present invention, the complex fermented extract of hibiscus, kelp, and mother-of-pearl is a complex fermented with Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain. It is characterized in that it is a fermented extract. The present inventors made intensive research efforts to further improve the skin condition improvement effect of Japanese radish, kelp, and mother-of-pearl extracts through natural product fermentation, and natural complex fermentation fermented with the Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain. It has been confirmed that the extract's skin condition improvement effect on skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing effect is more excellent.

In the cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant, and skin moisturizing of the present invention, the complex fermented extract of Japanese kelp, kelp, and mother-of-pearl increases the expression of type I procollagen gene, and MMP-1 (matrix metallo-proteinase 1) It is characterized in that it has the activity of improving skin elasticity or skin wrinkles by suppressing the expression of genes.

As used herein, the term "skin wrinkle" refers to fine lines caused by skin aging, and may be caused by genetic causes, a decrease in collagen present in the dermis of the skin, external environment, and the like. In the present invention, "skin wrinkle prevention or improvement" refers to suppressing or inhibiting the generation of wrinkles on the skin, or alleviating the already generated wrinkles.

The main function of skin fibroblasts is regeneration of damaged skin tissue through extracellular matrix synthesis and proliferation. They lower the physiologically active function of dermal fibroblasts in the dermal layer. Type I collagen, which accounts for more than 95% of the skin extracellular matrix, plays a very important role in the physiological activity of fibroblasts through interaction and signal exchange with cell adhesion molecules and cytoskeleton, etc. When the synthesis of cause a vicious cycle.

Therefore, it can be seen that collagen of dermal fibroblasts is directly related to skin regeneration, skin elasticity, skin wrinkle formation, and tissue repair or regeneration during skin damage.

That is, when the synthesis of collagen or procollagen in skin fibroblasts is promoted or the synthesis of matrix metallo-proteinase (hereinafter referred to as 'MMP') that decomposes collagen is inhibited, skin elasticity improvement, skin regeneration, It is possible to obtain effects such as skin wrinkle improvement, wound healing, repair and regeneration of damaged skin tissue, and prevention of skin aging.

In an embodiment of the present invention, the complex extract or ferment extract of H. hemp, kelp, and mother-of-pearl increases the expression of type I procollagen gene, and suppresses the expression of MMP-1 (matrix metallo-proteinase 1) gene to improve skin elasticity or It was confirmed that it had the activity of improving skin wrinkles, and it could be confirmed that there was a synergistic effect due to the combination of Japanese apricots, kelp, and mother-of-pearl. In addition, it was confirmed that an additional effect occurred in the improvement of excellent skin elasticity or skin elasticity or skin wrinkles due to the fermentation treatment (see FIGS. 2 and 3 ).

In addition, the cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing of the present invention is characterized in that it has antioxidant activity to eliminate reactive oxygen species. In a preferred embodiment of the present invention, it was confirmed that the complex extract of H. shiitake, kelp, and mother-of-pearl and their complex fermented extract had antioxidant activity to scavenging active oxygen species in a concentration-dependent manner, and the effect was It was confirmed that there was a marked increase compared to the single extract of (see FIG. 4).

In addition, in the cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidation and skin moisturizing of the present invention, the complex extract of hibiscus, kelp, and mother-of-pearl or its fermented extract promotes hyaluronic acid synthesis to have skin moisturizing activity. do. The term "moisturizing the skin" means to increase the feeling of moisture in the skin and to maintain a moist state. In a preferred embodiment of the present invention, it was confirmed that the complex extract of H. kelp, kelp, and mother-of-pearl and their complex fermented extract increased the production amount of hyaluronan in a concentration-dependent manner (see FIG. 5 ).

The cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing of the present invention is derived from edible, naturally-derived plant material, is harmless to the human body, and does not impair the quality of the cosmetic material due to its addition, so it can be used in a cosmetic composition It is suitable for use with In particular, since it has functions related to skin wrinkle improvement, antioxidant and skin moisturizing, it is preferably included in the functional cosmetic composition.

The cosmetic composition of the present invention includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, mask pack, It may be prepared in any one formulation selected from soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser, but is not limited thereto.

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, and thickener. , a chelating agent, a colorant, a preservative, a fragrance, and the like may be appropriately mixed, but the present invention is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohard propellants such as, but not limited to, locarbon, propane/butane or dimethyl ether. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil, etc. can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, fatty acid ester of polyethylene glycol or sorbitan may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. are used as carrier components may be, but is not limited thereto. These may be used alone or in combination of two or more.

In the cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant, and skin moisturizing of the present invention, the complex fermented extract of hibiscus, kelp, and mother-of-pearl is specifically, in dry weight, 0.0001% to 50% by weight of the total weight of the cosmetic composition It may be included, and more specifically, it may be included in an amount of 0.0005 wt% to 10 wt%. Within the above range, there is an advantage of exhibiting excellent efficacy for skin elasticity or skin wrinkles improvement, antioxidant and skin moisturizing, and there is an advantage in that the formulation of the composition is stabilized.

According to another aspect of the present invention, the present invention is Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain fermented with a complex fermented extract of hibiscus, kelp, and mother-of-pearl as an active ingredient for skin elasticity or skin wrinkles A food composition for improving, antioxidant and skin moisturizing is provided.

In the food composition for skin elasticity or wrinkle improvement, antioxidant and skin moisturizing of the present invention, the complex extract is a complex fermented extract fermented with a Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain. .

The complex fermented extract fermented with the Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain and the complex extract of Japanese radish, kelp, and motherwort of the present invention has excellent activity in skin elasticity and skin wrinkle improvement, and at the same time has excellent antioxidant and antioxidant properties. It can be usefully used as a food composition for improving skin condition by showing an excellent effect in moisturizing the skin. The complex extracts of shiitake, kelp, and motherwort of the present invention, their complex fermented extracts, and the effects on "improving skin tightening or skin wrinkles" and "skin moisturizing" are the same as those described above. The food composition may be used in the form of health functional food, but is not limited thereto.

The food composition of the present invention may be included in the form of a fraction for a complex fermented extract of H. shiitake, kelp, and mother-of-pearl or a processed product thereof. In addition, the composition may include a food pharmaceutically acceptable food additive in addition to the active ingredient.

In the present invention, "food supplementary additive" refers to a component that can be supplementally added to food, and is added to the manufacture of health functional food of each formulation, and those skilled in the art can appropriately select and use it. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of the food additive of the present invention.

The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, the term “functionality” refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological actions with respect to the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.

In addition, if the formulation of the health functional food is also recognized as a health functional food, it may be manufactured without limitation. The food composition of the present invention can be prepared in various forms, and unlike general drugs, it uses edible seaweed as a raw material, so there are no side effects that may occur when taking food for a long period of time, and it is excellent in portability and reduces skin wrinkles. It can be taken as an adjuvant to enhance the anti-aging effect, to show an antioxidant effect, and to increase the moisture content of the skin.

There is no limitation in the form that the health functional food of the present invention can take, and it may include all foods in a conventional sense, and may be used in combination with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in food according to the selection of those skilled in the art. Examples of foods that can be added include 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 There are vitamin complexes and the like, and it can be prepared by adding to juice, tea, jelly, juice, etc. prepared by using the complex extract of shiitake, kelp, and mother-of-pearl according to the present invention or a complex fermented extract thereof as a main component. Also included are foods used as feed for animals.

The features and advantages of the present invention are summarized as follows:

(1) The present invention is Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain fermented with a complex fermented extract of hemp, kelp, and mother-of-pearl as an active ingredient Skin elasticity or skin wrinkle improvement, antioxidant and skin A moisturizing composition is provided.

(2) The composition of the present invention has no cytotoxicity, has an excellent effect on improving skin elasticity or skin wrinkles, and has an excellent effect on antioxidant and skin moisturizing, so as to improve skin elasticity or skin wrinkles in cosmetics and food fields, antioxidant And it can be usefully used for the purpose of moisturizing the skin.

(3) In addition, the composition of the present invention is not only very safe for the human body, but also has excellent stability.

1 is a graph showing the measurement of the cell viability of human skin fibroblasts according to the treatment of a complex extract of Japanese daisy, kelp, and mother-of-pearl or a complex fermented extract thereof.
2 is a graph showing changes in the expression of type I procollagen protein according to treatment with a complex extract of kelp and mother-of-pearl or their complex ferment extract in human fibroblasts.
3 is a graph showing the effect of inhibiting the expression of MMP-1 protein in human fibroblasts, a complex extract of kelp and mother-of-pearl, or a complex fermented extract thereof.
4 is a graph showing the measurement of the active oxygen species (ROS) scavenging ability of the complex extract of kelp and mother-of-pearl or their complex fermented extract.
Figure 5 is a graph showing the measurement of changes in the amount of hyaluronic acid (Hyaluronan) produced according to the treatment of the complex extracts of Japanese radish, kelp, and mother-of-pearl and their complex fermented extracts in human keratinocytes (HaCaT).

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and therefore, the scope of the present invention is not to be construed as being limited by these examples.

Example 1. Preparation of extracts of Japanese radish, kelp, and mother-of-pearl and their complexes

1-1. Manufacture of single extracts of shiitake, kelp, and mother-of-pearl

After pulverizing 100 g of dried shiitake, 3 L of purified water was used as a solvent, followed by heating under reflux for 12 hours, followed by cooling, followed by filtration with filter paper having a permeation size of 1.2 μm to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare a hibiscus extract (Preparation Example 1).

A kelp extract (Preparation Example 2) and a mother-of-pearl extract (Preparation Example 3) were prepared by using the same method as the method for obtaining the H. shiitake extract.

In addition, by mixing the same amount of the dried shiitake, kelp, and mother-of-pearl powder, a complex extract (Preparation Example 4) thereof was prepared.

1-2. Preparation of fermented extract

Lactobacillus brevis elastic001 strain (Accession No. KCTC 14209BP) was inoculated into 100 g of the complex extract of Hachibana, kelp, and motherwort of Preparation Example 4, and cultured at 30° C. for 15 days to carry out the fermentation process.

After adding 10 times the volume of 50% ethanol to the fermentation product, reflux extraction was performed at 60° C. for 6 hours, followed by filtration, and the supernatant was concentrated with a reduced pressure concentrator and used as a fermentation extract (Preparation Example 5).

Example 2. Cytotoxicity test according to the addition of plant extracts

Cells against normal human dermal fibroblasts (NHDF) of single and complex extracts (Preparation Examples 1 to 4) and their complex fermented extracts (Preparation Example 5) of Japanese radish, kelp, and motherwort prepared in Example 1 Toxicity was tested.

Cytotoxicity was measured using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assay. Methyl thiazol tetrazolium (MTT) assay is used to measure cell viability, which converts MTT into a purple-producing formazan stain. After 72 hours of incubation, the volume of medium was reduced to 1 ml and 100 μl of 1 mg/ml MTT was added to each wall. Thereafter, normal human dermal fibroblasts (NHDF) were cultured in the presence of _{5% CO 2} and 95% O _{2 at 37° C. for 2 hours.} The substrate-containing medium was removed and 1 ml of DMSO was added to each wall to dissolve the formazan crystals. In addition, the plate was shaken with an orbital shaker at room temperature for 30 minutes. Absorption of 100 μl aliquots was quantified by measuring absorption at 570 nm using a microplate reader (Molecular Devices E09090; San Francisco, CA, USA).

Cells of each component in cells (72 hours after treatment) treated with single and complex extracts (Preparation Examples 1 to 4) and their complex fermented extracts (Preparation Example 5) of H. kelp, kelp, and motherwort prepared in Example 1 To confirm toxicity, MTT analysis was performed.

As a result, as shown in FIG. 1, the treatment of single and complex extracts (Preparation Examples 1 to 4) and their complex fermented extracts (Preparation Example 5) of H. kelp, kelp, and motherwort did not show toxicity to human skin fibroblasts. , rather, it was confirmed that growth was promoted, and the effect of the complex extract was superior to that of the single extract (Fig. 1). In particular, the complex fermented extract fermented by inoculating the Lactobacillus brevis elastic001 strain was 111%, and 107%, respectively, at 10, and 100㎍/㎖ concentration, compared to the single extract of hibiscus; 107%, and 119%, respectively, at 10, and 100 μg/ml concentrations compared to kelp single extract; And it was confirmed that there was a 104% and 114% improved cell proliferation effect at 10 and 100 μg/ml concentrations, respectively, compared to a single extract of mother and child.

Example 3. Skin wrinkle and elasticity improvement activity

3-1. Promotes the synthesis of type 1 procollagen protein

The expression change of type 1 procollagen protein in normal human dermal fibroblast (NHDF) according to the treatment of the extracts prepared in Example 1 was measured.

For the experiment, 2 ml of DMEM culture solution was placed in a 40mm cell culture dish, and human skin fibroblasts ^{were inoculated at a concentration of about 1.2x10 5} , and then cultured at 37° C., 5% CO ₂ environment for 24 hours. Thereafter, the medium (10 and 100 μg/ml, respectively) containing the single and complex extracts (Preparation Examples 1 to 4) of Japanese radish, kelp, and motherwort prepared in Example 1, and their complex fermented extract (Preparation Example 5) ) and cultured for 3 days. Thereafter, the culture medium was harvested and centrifuged for 5 minutes at 4° C., 7,500 rpm. Protein expression level of type 1 procollagen (Procollagen Type IC Peptide EIA Kit, Takara, Shiga, Japan) using ELISA method Change was confirmed.

Referring to FIG. 2 , it was found that the expression level of type 1 procollagen protein was insignificant in the single extract of H. hemp, kelp, and mother-of-pearl compared to the untreated negative control group. On the other hand, it was confirmed that the complex extract (Preparation Example 4) and their complex ferment extract (Preparation Example 5) increased the production of type 1 procollagen protein in a concentration-dependent manner. It was observed that the experimental group treated with the complex extract of H. hemp, kelp, and motherwort in both the 10 and 100 μg/ml treatment groups showed a significantly improved effect of increasing the synthesis of type 1 procollagen protein compared to the single treatment group. , 181%, and 166% at concentrations of 10, and 100 μg/ml, respectively, compared to Japanese shiitake single extract; 178%, and 183%, respectively, at 10, and 100 μg/ml concentrations compared to kelp single extract; In addition, 192% and 196% increase in the expression level of type 1 procollagen protein was observed at 10 and 100 μg/ml concentrations, respectively, compared to a single extract of mother and child, and through this, a synergistic effect due to the combination of Japanese radish, kelp, and motherwort was able to confirm that there is

In addition, the effect of increasing the expression of type 1 procollagen protein of the ferment extract of the Lactobacillus brevis elastic001 strain was observed to be more excellent, respectively, 132% at 10 and 100 μg/ml concentration compared to the complex extract treatment group without fermentation treatment, and A 121% increase in the expression level of type 1 procollagen protein was observed.

Through the above results, the complex extract of Japanese radish, kelp, and mother-of-pearl of the present invention acts on the TGF-β/Smad signaling system to promote the production of type 1 procollagen, effectively prevent wrinkle formation, and It was confirmed that there was an effect of improving wrinkles and skin elasticity, and it was confirmed that there was a synergistic effect due to the combination of shiitake, kelp, and mother-of-pearl. In addition, it was confirmed that the excellent effect of increasing the expression of type 1 procollagen protein due to the fermentation process was additionally generated.

3-2. Inhibition of MMP-1 protein synthesis

The expression change of MMP-1 protein in normal human dermal fibroblast (NHDF) according to the treatment of the extracts prepared in Example 1 was measured.

For the experiment, 2 ml of DMEM culture solution was placed in a 40mm cell culture dish, and human skin fibroblasts ^{were inoculated at a concentration of about 1.2x10 5} , and then cultured at 37° C., 5% CO ₂ environment for 24 hours. Thereafter, the medium (10 and 100 μg/ml, respectively) containing the single and complex extracts (Preparation Examples 1 to 4) and their complex fermented extracts (Preparation Example 5) of Japanese radish, kelp, and motherwort prepared in Example 1 were used. It was replaced and cultured for 3 days. After that, the culture medium was harvested and centrifuged for 5 minutes at 4° C., 7,500 rpm, and MMP-1 (Human Total MMP-1 kit, R&D Systems, Inc., Minneapolis, MN, USA) using an ELISA method. ) of the protein expression level was confirmed.

Referring to FIG. 3, in the human skin fibroblast experiment, the complex extract (Preparation Example 4) and their complex fermented extract (Preparation Example 5) of H. kelp, kelp, and motherwort inhibited the production of MMP-1 protein in a concentration-dependent manner and was found to be It was observed that the experimental group treated with the complex extract of H. shiitake, kelp, and motherwort in both the 10 and 100 μg/ml treatment groups exhibited significantly improved MMP-1 protein production inhibitory effect compared to the single treatment group, 27%, and 25%, respectively, at 10, and 100 μg/ml concentrations compared to Japanese shiitake single extract; 32%, and 34%, respectively, at 10, and 100 μg/ml concentrations compared to kelp single extract; In addition, 25% and 30% inhibition of MMP-1 protein expression levels were observed at concentrations of 10 and 100 μg/ml, respectively, compared to a single extract of mother and child, and through this, a synergistic effect due to the combination of H. It could be confirmed that there is (FIG. 3).

In addition, it was observed that the MMP-1 protein expression inhibitory effect of the fermented extract of the Lactobacillus brevis elastic001 strain of the present invention was further increased compared to the complex extract of Hachibana, kelp, and motherwort, compared to the complex extract treated group without fermentation treatment 10, and 11% and 17% inhibition of MMP-1 protein expression levels, respectively, were observed at 100 μg/ml concentration.

Through the above results, the complex extract of shiitake, kelp, and mother-of-pearl of the present invention can be effectively used to prevent and improve skin wrinkles by effectively inhibiting the production of MMP-1 protein, which is the cause of wrinkles, It was confirmed that there was a synergistic effect due to the combination of kelp, and mother-of-pearl. In addition, it was confirmed that the excellent effect of inhibiting MMP-1 protein expression due to the fermentation process was additionally generated.

Example 4. Antioxidant activity

Experiments to confirm the scavenging ability of ROS (Reactive Oxygen Species) of the complex fermented extract of Japanese daisy, kelp, and mother-of-pearl fermented with Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) strain in human skin fibroblasts carried out.

The experiment was conducted by irradiating human skin fibroblasts with UVB (UVB Peak range: 312nm, UV dose: 144mJ/cm ² ), and then using the single extracts (Preparation Examples 1 to 3) and of the complex extract (Preparation Example 4), and their complex ferment extract (Preparation Example 5) were diluted with serum-free DMEM medium, treated at concentrations of 10 and 100 µg/ml, respectively, and incubated for 30 minutes, excitation Fluorescence was measured at 480 nm and emission 530 nm, and DCF fluorescence intensity was compared. Statistical processing was performed using the GraphPad Prism program, and the significance of the mean value was investigated to a limit of less than 5% using ANOVA (one-way analysis of variance).

As a result, referring to FIG. 4 , it was measured that the experimental groups treated with a single extract of Japanese radish, kelp, and mother-of-pearl exhibited 45%, 49%, and 51% reactive oxygen species scavenging ability at a concentration of 100 μg/ml, respectively. It was measured that the antioxidant effect of the components was not very high. On the other hand, it was observed that 71% and 65% of active oxygen species scavenging ability was measured at 10 and 100 μg/ml concentrations of the complex extracts of Japanese radish, kelp, and motherwort in which these three components were combined, and a synergistic effect of the combination was observed. .

In addition, in the test group treated with the complex fermented extract of Japanese daisy, kelp, and mother-of-pearl fermented with Lactobacillus brevis elastic001 strain, more improved reactive oxygen species scavenging activity was observed compared to the complex extract that was not fermented, specifically 10 and At a concentration of 100ug/ml, 28% and 34% more improved reactive oxygen species scavenging ability was measured, respectively, compared to the test group not subjected to fermentation.

Example 5. Skin moisturizing activity

In order to verify the skin moisturizing effect of the complex extracts of shiitake, kelp, and mother-of-pearl of the present invention and their complex fermented extracts, their effect on promoting hyaluronan synthesis was tested.

Hyaluronic acid (HA, Hyaluronan, Hyaluronic acid) is mainly synthesized by epidermal keratinocytes and dermal fibroblasts, and is a substance that plays an important role in creating an environment in which cells can function normally by combining with water. A decrease in hyaluronic acid in the skin is one of the direct causes of a decrease in skin elasticity and a decrease in moisture content. Therefore, in this experiment, the amount of hyaluronic acid produced in human keratinocytes was confirmed.

Human immortalized keratinocytes (HaCaT, Human immortalized keratinocytes) were placed in 1 ml of a culture medium in a 24-well cell culture dish ^{and inoculated at a concentration of about 1×10 5} , 37°C, 5% CO ₂ environment for 24 hours. cultured. Thereafter, the single extracts (Preparation Examples 1 to 3) and their complex extracts (Preparation Example 4), and their complex fermented extracts (Preparation Example 5) prepared according to Example 1 were prepared It was replaced with a medium containing 10 and 100 μg/ml, respectively, and cultured for 24 hours. Thereafter, the culture medium was harvested and centrifuged for 5 minutes at 4° C., 7,500 rpm, and the expression level of Hyaluronan (Hyaluronan kit, R&D Systems, Inc., Minneapolis, MN, USA) was performed using an ELISA method. Change was confirmed.

As a result, referring to FIG. 5 , it was found that there was no significant difference in the production amount of hyaluronic acid (Hyaluronan) between the untreated negative control group and the untreated test group treated with a single extract of seaweed, kelp, and motherwort. In contrast, in the test group treated with the three types of complex extracts of kelp, kelp, and motherwort, it was confirmed that the production amount of hyaluronan increased in a concentration-dependent manner, and 10 and 100ug/ml compared to the untreated negative control group. It was confirmed that the production amount of hyaluronic acid increased by 171% and 202%, respectively, at the concentration of .

In addition, in the test group treated with the complex fermented extract of Japanese daisy, kelp, and motherwort fermented with Lactobacillus brevis elastic001 strain, an increased amount of hyaluronic acid production was observed compared to the complex extract that was not fermented. It was confirmed that the production amount of hyaluronic acid increased by 197% and 247%, respectively, at the concentrations of 10 and 100ug/ml compared to the treatment negative control group, and it was confirmed that there was a very good effect in moisturizing the skin.

Through the results as described above, the complex extract of Japanese radish, kelp, and mother-of-pearl of the present invention and their complex fermented extract have no cytotoxicity, and improve skin elasticity or It significantly contributes to skin wrinkle improvement, effectively increases the production of hyaluronic acid, so it is effective in moisturizing the skin, and it can be confirmed that it can be used very effectively to improve skin condition by showing antioxidant activity.

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Korea Institute of Biotechnology and Biotechnology KCTC14209 202000608

Claims (6)

Lactobacillus brevis elastic001) (Accession No. KCTC 14209BP) A cosmetic composition for improving skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing, comprising a complex fermented extract of shiitake, kelp, and mother-of-pearl fermented with a strain as an active ingredient. According to claim 1,
The composition increases the expression of the type I procollagen gene and suppresses the expression of the MMP-1 (matrix metallo-proteinase 1) gene to improve skin elasticity or skin wrinkles, characterized in that it has skin elasticity or skin wrinkle improvement activity; A cosmetic composition for antioxidant and skin moisturizing. According to claim 1,
The composition is a cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing, characterized in that it has antioxidant activity to eliminate reactive oxygen species (ROS). According to claim 1,
The composition promotes the synthesis of hyaluronic acid, and the skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing cosmetic composition, characterized in that it has a skin moisturizing activity. According to claim 1,
The composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, mask pack, soap, cleansing A cosmetic composition for skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing, characterized in that the formulation is any one selected from foam, cleansing lotion, cleansing cream, body lotion and body cleanser. Lactobacillus brevis elastic001 (Accession No. KCTC 14209BP) A food composition for improving skin elasticity or skin wrinkle improvement, antioxidant and skin moisturizing, comprising a complex fermented extract of shiitake, kelp, and mother-of-pearl fermented with a strain as an active ingredient.

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