KR102145030B1 - A cosmetic composition for improving skin barrier function power containing an extract of fermented omija as an effective ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient, and specifically, a cosmetic composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient, a health functional food composition and a quasi-drug composition Provides. The cosmetic composition, health functional food composition and quasi-drug composition of the present invention has low cytotoxicity, inhibits oxidation and inflammation of the skin, and improves skin barrier function, thereby enhancing skin moisturizing power, preventing skin troubles, or skin aging. It has the effect of preventing.

Description

A cosmetic composition for improving skin barrier function containing fermented Schisandra chinensis extract and as an active ingredient {A COSMETIC COMPOSITION FOR IMPROVING SKIN BARRIER FUNCTION POWER CONTAINING AN EXTRACT OF FERMENTED OMIJA AS AN EFFECTIVE INGREDIENT}

The present invention relates to a fermented Schisandra chinensis extract and a cosmetic composition for improving skin barrier function containing the same as an active ingredient.

Skin is an organ that is always in contact with the external environment. It protects the human body from external physical damage and chemicals, prevents bacteria, fungi, and viruses from invading the skin, and acts as a protective barrier against moisture loss. The skin is composed of three layers: the epidermis, the dermis, and the subcutaneous fat, and the epidermis is the thinnest layer among the three layers above, and plays an important role in moisturizing and protecting the skin. In particular, the skin has elasticity and is softened by moisture present in the stratum corneum of the epidermis. It is known that moisture content of 10% or more is essential to maintain the elasticity of the stratum corneum.

Skin hydration is determined by the equilibrium of water transfer from the epidermis to the dermis due to osmotic pressure and the loss of water from the epidermis due to evaporation. Clinically, in dry skin, the skin surface is dry and elastic, and scaling is easy and rough. In addition, itchy symptoms may appear, and in some cases, symptoms such as dryness and cracking may be accompanied. The level of moisturizing in the epidermis is highly dependent on lipids, corneodesmosom, and natural moisturizing factor (NMF) between keratinocytes.

Involucrin is a type of cytoplasmic protein synthesized in squamous epithelial cells of the human body. In normal epidermis, involucrin begins to appear from the upper parietal layer and granular layer, and after synthesis, it is located near the cell membrane and is produced by trasnglutamidase. It is a major structural constituent protein of the cornified cell envelope, which is bound to the membrane protein and acts as a support, and maintains skin moisturizing power by performing a skin barrier function.

Filaggrin is a kind of epidermal protein, which not only performs the function of maintaining moisture and moisturizing ingredients in the skin, but is also an essential protein to inhibit the penetration of bacteria and toxins from the outside. First, it is secreted in the form of profilaggrin, a precursor of filaggrin, and then decomposed into filaggrin monomer, which aggregates keratin intermediate fine fibers in the stratum corneum to form an insoluble keratin matrix. Using this substrate as a skeleton, a cornified cell envelope is formed just below the cell membrane of keratinocytes. And filaggrin is further decomposed into NMF, a hydrophilic amino acid. As a powerful moisturizer, NMF plays an important role in supplying moisture to the stratum corneum and lowers the pH of the stratum corneum to have an antibacterial effect.

In the present invention, a cosmetic composition capable of improving skin barrier function by increasing the expression of involucrin and filaggrin proteins using a fermented extract derived from Schisandra chinensis, a natural substance, health functional food composition and quasi-drug It was intended to provide a composition.

An object of the present invention is to provide a cosmetic composition for improving the skin barrier function containing the fermented Schisandra chinensis extract as an active ingredient.

Another object of the present invention is to provide a health functional food composition for improving skin barrier function containing fermented Schisandra chinensis extract as an active ingredient.

Another object of the present invention is to provide a quasi-drug composition for improving skin barrier function containing fermented Schisandra chinensis extract as an active ingredient.

Another object of the present invention is to provide a method for preparing a fermented Schisandra chinensis extract for improving skin barrier function.

In order to achieve the above object, the present invention provides a cosmetic composition for improving skin barrier function, containing a fermented Schisandra chinensis extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a health functional food composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a quasi-drug composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient.

In order to achieve another object of the present invention, the present invention is a Pichia kudriavzevii (Pichia kudriavzevii), Saccharomycopsis fibuligera (Saccharomycopsis fibuligera), Lactobacillus buchneri (Lactobacillus buchneri), Schisandra fermentation for improving skin barrier function, comprising inoculating and fermenting at least one strain selected from the group consisting of Lactobacillus paracasei sub. Tolerans and Lactobacillus harbinensis It provides a method for producing an extract.

According to the present invention, the cosmetic composition, health functional food composition and quasi-drug composition for improving skin barrier function containing Schisandra chinensis fermentation extract as an active ingredient has low cytotoxicity, inhibits oxidation and inflammation of the skin, and improves skin barrier function. By doing so, there is an effect of enhancing skin moisturizing power, preventing skin troubles, or preventing skin aging.

1 is a graph showing cell viability (A) and NO production (B) when RAW264.7 cells were treated with Schisandra fermented extract.
2 is a graph showing mRNA (C) expression of inflammatory cytokines IL-6 (A), IL-β (B) and TNF-α when treated with Schisandra chinensis ferment extract on RAW264.7 cells. .
Figure 3 shows that when the Schisandra chinensis fermentation extract is treated with UVB-irradiated HaCaT (keratinocytes), the decrease in the expression of involukrin, filaggrin and mRNA is suppressed.
Figure 4 shows the increase in ERK-1/2 protein phosphorylation (activation) in HaCaT cells irradiated with UVB 300mJ/cm ² , 5h of the Schisandra ferment extract, and the increase in c-Fos protein and mRNA expression.
FIG. 5 shows the results of a hepatotoxicity experiment due to a reduction in liver size caused by apoptosis of hepatocytes and necrosis caused by a hepatotoxic drug and an experiment result when the fermented Schisandra chinensis extract was treated.
6 shows the results of a hepatotoxicity experiment due to a reduction in liver size caused by hepatocyte death and necrosis caused by a hepatotoxic drug, and observation results based on GFP fluorescence when treated with a fermented Schisandra chinensis extract.
FIG. 7 shows the results of quantitatively measuring protein leakage through the intestine by measuring fluorescence released after inducing intestinal damage of zebrafish and experimental results (absorbance) when treated with Schisandra fermented extract.
Figure 8 shows the result of quantitatively measuring the protein outflow through the intestine by measuring the fluorescence emitted after inducing intestinal damage of zebrafish and the experimental result (% comparison) when treated with Schisandra fermented extract.

Hereinafter, the present invention will be described in more detail.

Involucrin is a type of cytoplasmic protein synthesized in squamous epithelial cells of the human body. In normal epidermis, involucrin begins to appear from the upper parietal layer and granular layer, and after synthesis, it is located near the cell membrane and is produced by trasnglutamidase. It is the main structural constituent protein of the cornified cell envelope, which is bound to the membrane protein and acts as a support.

Filaggrin is a kind of epidermal protein, which not only performs the function of maintaining moisture and moisturizing ingredients in the skin, but is also an essential protein to inhibit the penetration of bacteria and toxins from the outside. First, it is secreted in the form of profilaggrin, a precursor of filaggrin, and then decomposed into filaggrin monomer, which aggregates keratin intermediate fine fibers in the stratum corneum to form an insoluble keratin matrix. Using this substrate as a skeleton, a cornified cell envelope is formed just below the cell membrane of keratinocytes. And filaggrin is further decomposed into NMF, a hydrophilic amino acid. As a powerful moisturizer, NMF plays an important role in supplying moisture to the stratum corneum and lowers the pH of the stratum corneum to have an antibacterial effect.

On the other hand, involucrin and pilargreen, which are known to have the function of maintaining and strengthening the skin barrier function, are natural moisturizing factors related to moisturizing and hydration of the skin in keratinocytes, and the fermented Schisandra chinensis extract according to the present invention is involucrin ( By increasing the expression of involucrin) and filaggrin proteins, it improves skin barrier function, strengthens skin moisturizing power, prevents skin troubles, or prevents skin aging.

Accordingly, according to an embodiment of the present invention, there is provided a cosmetic composition for improving skin barrier function, containing a fermented Schisandra chinensis extract as an active ingredient. In addition, according to an embodiment of the present invention, specifically, the cosmetic composition may have one or more uses selected from the group consisting of enhancing skin moisturization, preventing skin troubles, and preventing skin aging.

In the present invention, Schisandra chinensis is a fruit opened on a vine-like deciduous tree of the family Schisandra chinensis, and is called Schisandra because it has five flavors: sweet, bitter, sour, spicy and salty. Schisandra chinensis is generally known to be effective in improving blood flow, high blood pressure, stroke, cardiovascular disease prevention, immunity improvement, diabetes prevention, liver function improvement, rejuvenation, and respiratory disease improvement.

In the present invention, the fermented Schisandra chinensis extract may be obtained by extraction and fermentation from various organs or parts of Schisandra chinensis, and as an example, it may be a fermented product of hot water extract of Schisandra chinensis obtained by fermenting the extract obtained by hot water extraction of Schisandra chinensis with a specific strain. .

Specific strains used in one embodiment of the present invention are Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus paraca J. Tolerance (Lactobacillus paracasei sub. Tolerans) and Lactobacillus habinensis (Lactobacillus harbinensis) may be one or more strains selected from the group consisting of.

On the other hand, the cosmetic composition according to an embodiment of the present invention is not limited to being prepared in any formulation commonly prepared in the art, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, It may be formulated as a lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. In more detail, it can be applied to manufacture functional cosmetics that are basic cosmetics such as lotion, nutritional cream, moisture cream, essence, eye cream, lotion, pack, and essence.

On the other hand, the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the fermented extract as an active ingredient, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances. It may include conventional adjuvants and carriers.

On the other hand, according to an embodiment of the present invention, the fermented Schisandra chinensis extract may be contained in an amount of 0.1 to 5% by weight based on the total weight of the cosmetic composition, and has an effect of improving skin barrier function within the weight% range, and the weight If it is less than the% range, there may be a problem that the effect of improving the skin barrier function is insignificant.

On the other hand, according to an embodiment of the present invention, it provides a health functional food composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient. Like the cosmetic composition described above, the health functional food composition may also have one or more uses selected from the group consisting of skin moisturizing enhancement, skin trouble prevention, and skin aging prevention.

In the health functional food composition according to an embodiment of the present invention, there is no particular limitation on the type of health functional food, for example meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gums, dairy products , Various soups, beverages, teas, drinks, alcoholic beverages, vitamin complexes, etc., and may include all foods in a conventional sense.

On the other hand, the food composition may include ingredients commonly used in health functional food compositions in addition to the fermentation extract, such as nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids , A protective colloidal heavy agent, a pH adjuster, a stabilizer, a preservative, a glycerin, an alcohol, a carbonation agent used in carbonated beverages, or a combination thereof.

On the other hand, according to an embodiment of the present invention, there is provided a quasi-drug composition for improving skin barrier function containing a fermented Schisandra chinensis extract as an active ingredient. Like the cosmetic composition described above, the quasi-drug composition may also have one or more uses selected from the group consisting of skin moisturizing enhancement, skin trouble prevention, and skin aging prevention.

The term "quasi-drug" as used in the present invention refers to items that are less effective than medicines among items used for diagnosis, treatment, improvement, alleviation, treatment or prevention of diseases of humans or animals, for example According to the report, quasi-drugs exclude items used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases of humans or animals, and products with minor or no direct action on the human body.

The quasi-drug composition is used for the purpose of improving skin barrier function, and more specifically, for enhancing skin moisturization, preventing skin troubles and preventing skin aging, and is not particularly limited in its formulation, for example, softening lotion, nutrition. It may be a composition having a formulation of lotion, massage cream, nutrition cream, pack, mask pack, mask sheet, gel or skin adhesive type cosmetic, and also a transdermal dosage form such as lotion, ointment, gel, cream, patch or spray It may be a composition having.

Meanwhile, in addition to the fermentation composition, the quasi-drug composition may include, for example, a thickener, a stabilizer, a solubilizing agent, a conventional auxiliary agent such as vitamins, pigments, and flavors, and a carrier as appropriate.

On the other hand, according to an embodiment of the present invention, Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus para Casei Tolerance (Lactobacillus paracasei sub. Tolerans) and Lactobacillus harbinensis (Lactobacillus harbinensis) at least one strain selected from the group consisting of inoculation and fermentation of one or more strains, comprising the step of fermenting Schisandra chinensis fermentation extract Provides.

Specifically, the Schisandra chinensis extract according to an embodiment of the present invention prepares Schisandra chinensis by-products, and then adds 10 times of purified water to the weight ratio (wt%) of the Schisandra chinensis by-product, and hot water for 4 to 30 hours at a temperature condition of 70 to 90°C. It may be extracted. If the temperature condition in the above step is less than 70° C., there may be a problem that it is difficult to effectively perform extraction, and if the temperature condition is more than 90° C., there is a problem that the active ingredient may be destroyed.

On the other hand, the fermentation may be performed by inoculating microorganisms in a number of bacteria of 2.0 to 7.0 at an absorbance value of OD 600 and performing for 24 to 72 hours at 25 to 37 °C. Specifically, when fermentation is performed with bacteria less than 2.0 at OD 600 absorbance, there is a problem that bacteria may not grow, and when fermentation is performed with more bacteria than 7.0 at OD 600 absorbance, contaminated bacteria may be inoculated. There is this.

Meanwhile, according to an embodiment of the present invention, the fermentation may be performed for 24 to 72 hours. Specifically, when fermentation is performed for less than 24 hours, there may be a problem that it is difficult to effectively perform fermentation, and when fermentation is performed for more than 72 hours, contamination may be a problem.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Example

Example 1. Preparation of fermented Schisandra chinensis extract

100 g of the Schisandra chinensis by-product remaining after being used in the production of Schisandra chinensis was prepared, and then the Schisandra chinensis by-product was mixed with 1000 g of water and then hot water extracted at 80° C. for 24 hours to obtain a Schisandra chinensis hot water extract.

The Schisandra chinensis ferment extract was obtained by inoculating a strain of Lactobacillus paracasei sub. Tolerans to the hot water extract. During the fermentation, the number of bacteria was inoculated with an OD 600 absorbance value of 4.0 to 6.0, and a Schisandra chinensis by-product was prepared at a ratio of 10 wt%, and 1 to 2% of the culture medium (OD 600 absorbance value 0.1) was inoculated, and 48 in an incubator. Incubated for hours. The culture broth was centrifuged for 30 minutes to collect the supernatant without bacteria, and the collected supernatant was filtered with a filter (5 μm pore size) to finally obtain a fermented Schisandra chinensis extract.

Example 2. Schisandra chinensis fermentation extract 30% ethanol extract

The fermented Schisandra chinensis extract prepared according to Example 1 was extracted using PB-600 adsorption resin to obtain a 30% ethanol extract of Schisandra chinensis fermentation extract. During the extraction, 300 g of PB-600 adsorption resin was prepared, and 1000 ml of Schisandra fermented extract was poured. Then, 500ml of DW, 10% ethanol, and 30% ethanol were sequentially flowed. Finally, 30% ethanol extract was obtained from the fermented Schisandra chinensis extract by pouring 30% ethanol.

[Experiment 1. Cytotoxicity test of fermented Schisandra chinensis extract]

Macrophage cell line RAW264.7 was pre-sale from Korea Cell Line Bank (KCLB, Seoul, Korea) and 5% CO using DMEM medium (Welgene, Gyeongsan, Korea) added with 10% FBS and 1% antibiotics (penicillin/streptomycin). ₂ was used after passage 2 to 3 times in an incubator at 37°C.

Cell viability measurement

To measure the cell viability, Raw 264.7 cells were dispensed into a 48 well plate at 3×10 ⁵ cells/well, and 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 was 0.1, 0.25, 0.5, 0.75 and 1 mg/mL. 2 hours after treatment, LPS 1 ug/mL was treated and cultured. After 24 hours, MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) was added, and DMSO (dimethyl sulfoxide) solution was added to the formazan formed after 4 hours reaction. Then, the absorbance was measured at 570 nm (see FIG. 1).

NO (Nitric Oxide) measurement

In order to measure the amount of NO produced in cells, Raw 264.7 cells were treated with 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2, stimulated with 1 ug/mL LPS, and cultured for 24 hours, and then Griess equivalent to the cell culture solution. Reagents (1% sulfanilamine, 0.1% naphthylethylene diamine di-hydrochloride, 2% phosphoric acid) were added and reacted, and the absorbance was measured at 540 nm. The concentration of NO was calculated based on the quantification curve of the NaNO ₂ standard solution (see FIG. 1).

Cytokine measurement

In order to measure the expression levels of the inflammatory cytokines IL-6, IL-1β, and TNF-α mRNA generated in cells, Raw 264.7 cells were treated with 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2, and LPS 1ug/ After stimulation with mL and culture for 24 hours, a culture supernatant was obtained. The cytokine concentration was measured using an ELISA (enzyme-linked immunosorbent assay) kit. A 96 well plate was coated with 100 μL of a capture antibody for each cytokine on a 96 well plate, and then left at 4° C. for 12 hours. The plate was washed with PBS (phosphate-buffered saline) containing 0.05% tween-20™, and then blocked with PBS containing 10% FBS for 1 hour. After washing again, the obtained cell culture solution was added and reacted at room temperature for 2 hours. Each well was washed again, a detection antibody and an enzyme reaction solution (SAv-HRP reagent) were added, followed by addition of the substrate ABTS solution to react. The color development reaction was measured at 405 nm using an ELISA reader (see FIG. 2).

Experiment result

As a result of measuring the inhibitory effect on NO production of the RAW264.7 macrophage line according to the content of 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2, it was found that NO production decreased as the content of fermented Schisandra chinensis extract at the same concentration increased. appear. Therefore, it was found that the anti-inflammatory effect was increased in proportion to the concentration of the fermented Schisandra chinensis extract. On the other hand, it was confirmed that the NO production inhibitory effect of the Schisandra fermented extract exhibited 87% NO production inhibitory effect at the maximum concentration of 0.5mg/ml without cytotoxicity (see FIG. 1).

On the other hand, it was confirmed that the amount of mRNA expression of the inflammatory cytokines IL-6, IL-1β and TNF-α significantly decreased with increasing concentration of the 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 (Fig. 2).

[Experiment 2. Schisandra chinensis fermentation extract promotes skin barrier function maintenance factor protein and mRNA expression]

Cell proliferation and viability measurement

Cytotoxicity test was performed by cell count and MTS assay method, specifically, the concentration of 30% ethanol extract of the fermented extract of Schisandra chinensis of Example 2 was treated at 25, 50, and 100 ug/ml concentrations, respectively, and confirmed with a cell counter. (See Fig. 3).

Western blot experiment

In order to confirm the expression of involukrin and filaggrin proteins in HaCaT cells irradiated with UVB, it was confirmed through Western blotting.

Specifically, 24 hours after inoculation of human keratinocytes HaCaT, 30% ethanol extract of Schisandra chinensis fermentation extract of Example 2 was cultured in DMEM medium, 37° C., 5% CO ₂ by adding EGF as a positive control, respectively. . After culturing for 24 hours, the cells were recovered, crushed, and only protein was extracted and stored at -40°C. The extracted protein was quantified through BCA assay. Each sample was diluted with 30 g of protein, and electrophoresed on a 12% SDS-gel, and then transferred to a PVDF membrane. After reacting with a primary antibody (rabbit anti-aquaporins-3 antibody), the remaining antibody was washed away and reacted with a secondary antibody (rabbit HRP-coupled anti-IgG antibody). After removing the remaining antibody, a reagent (peroxidase substrate and chromogen solution) that reacts with the antibody to cause a color reaction was added. The colored band was measured through image analysis (BioRad, Universal Hood, USA) (see FIG. 3).

RT-PCR experiment

First, 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 was added to human HaCaT (Human keratinocytes) keratinocytes and cultured in DMEM medium, 37°C, 5%, CO ₂ . After 24 hours incubation, cells were recovered, RNA was extracted, and PCR was performed using reverse transcriptase and cDNA (complementary DNA). GAPDH was used as a standard. The band expressed through the electrophoresis of the agarose gel was measured through image analysis (BioRad, Universal Hood, USA) (see FIG. 4).

Experiment result

In the case of the cell proliferation rate and cell viability according to the concentration of the 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2, it was confirmed that the concentration-dependent increase (see Fig. 3).

On the other hand, the expression levels of involucrine and filaggrin proteins and mRNA in UVB-unirradiated HaCaT cells did not change with the addition of Schisandra chinensis fermentation extract, but under UVB irradiation conditions, involucrine and filaggrin proteins and mRNA were also concentration-dependent. It was confirmed that the expression was increased (see FIG. 3).

In addition, when the 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 was treated at a concentration of 100 ug/ml during UVB irradiation, specifically UVB 300 mJ/cm ² , ERK-1/2 protein in HaCaT cells irradiated for 5 h It was confirmed that phosphorylation (activation) increases and c-Fos protein and mRNA expression increase (see FIG. 4).

[Experiment 3. Liver protection and gastrointestinal protection experiment of fermented Schisandra chinensis extract]

Liver protection experiment

Based on the reduction in liver size caused by hepatotoxic drug apoptosis and necrosis, based on the standard data of liver size change by acetaminophen, a hepatotoxic drug, the fermented extract of Schisandra chinensis of Example 2 The 30% ethanol extract was pretreated and then treated with acetaminophen to confirm the hepatoprotective effect of the 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2.

Specifically, for the adult zebrafish used in the test, an AB strain with clear source and a healthy LFABP-GFP homozygous transformant established through genetic engineering was used.

LFABP-GFP embryos were incubated 48 hours after fertilization, and then 5 individuals per well were transferred to a 24 well plate. The volume of the well was adjusted to be 1 ml. The experiment was conducted in the following four groups.

Control group: no treatment group

Experimental group 1: acetaminophen treatment group

Experimental group 2-4: acetaminophen treatment group after pretreatment of 30% ethanol extract of fermented extract of Schizandra chinensis of Example 2

Specifically, the concentration of the drug to be treated for each well to be tested was individually entered, and 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 was added according to the concentration on the second day, and the added drug was carefully rotated in a circular shape. It was made to mix evenly. Next, it was transferred to an incubator and incubated for 24 hours, and then acetaminophen was added on the third day. The size and shape of the liver were measured by observing with a fluorescence microscope on the 4th and 5th days, and mean±SD results were obtained by averaging the data obtained from 10 animals (see FIGS. 5 and 6).

Gastrointestinal protection experiment

KR68766 is a fluorescent substance that binds to albumin, and it is intended to quantitatively measure protein leakage through the intestine by measuring fluorescence that flows out to the medium when intestinal damage is caused after staining of zebrafish embryos.

Specifically, adult zebrafish were reared at 28.5° C. for 14 hours in the daytime and 10 hours in the night. In the afternoon of the previous day, a pair of males and females were placed in a dedicated mating cage, and the fertilized eggs were collected the next morning. The collected fertilized eggs were transferred to a petri dish and generated in an incubator at 28.5℃, and the development process was observed through a dissecting microscope.

15 embryos per well were placed on the 3rd day after fertilization in a 24-well plate. After dividing into an experimental group and a control group, the experimental group was pretreated with a 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 for 24 hours. The next day, after staining with KR68766, indomethacin was treated for 24 hours to induce intestinal damage. The next day, a medium was taken and fluorescence absorbance was measured (see FIGS. 7 and 8).

Experiment result

When the 30% ethanol extract of the fermented Schisandra chinensis extract of Example 2 was treated, it was confirmed that the liver damage protection effect by acetaminophen was exhibited at a drug concentration of 10 ug/ml, and the liver protection effect increased as the concentration increased. I could confirm. However, the liver size decreased in the 100 ug/ml alone treatment group, and all of the 100 ug/ml + acetaminophen group showed lethality (see FIGS. 5 and 6).

In the case of gastrointestinal protective effect, similar to the liver toxicity test, the protective effect increased as the drug concentration increased, but there was no significant difference at the concentration of 1 ug/ml, and drug toxicity was observed in the group treated with 100 ug/ml alone, and 100 ug/ml + Indomethacin group all showed lethality (see FIGS. 7 and 8).

Therefore, it was judged to show a significant liver and gastrointestinal protective effect at a drug concentration of 1-10 ug/ml.

Formulation Example 1. Flexible lotion

Formulation examples of flexible lotion were prepared as shown in Table 1 below according to a conventional method.

Formulation Example 2. Essence

Formulation examples of the essence were prepared as shown in Table 2 below according to a conventional method.

Formulation Example 3. Pack

Formulation examples of the pack were prepared as shown in Table 3 below according to a conventional method.

According to the experimental examples of the present invention, in the case of the fermented extract of Schisandra chinensis of the present invention according to the examples, expression of involucrin and filaggrin proteins known to have a function of maintaining and strengthening skin barrier function It was confirmed that there is an effect of improving the skin barrier function by increasing the skin barrier function. Accordingly, it has low cytotoxicity, suppresses oxidation and inflammation of the skin, and improves the skin barrier function, thereby enhancing skin moisturizing power and reducing skin troubles. It was confirmed that it can be used as a cosmetic ingredient due to the effect of preventing or preventing skin aging.

Claims (12)

Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus paracasei tolerance and Lactobacillus paracasei sub. At least one strain selected from the group consisting of Nensis (Lactobacillus harbinensis) was inoculated with a number of bacteria of 2.0 to 7.0 at an absorbance value of OD600 and fermented for 24 to 72 hours at 25 to 37°C. It contains 0.1 to 5% by weight based on,
The Schisandra chinensis fermentation extract is a fermented product of Schisandra chinensis extract obtained by adding purified water 10 times the weight of Schisandra chinensis to the Schisandra chinensis by-product, and extracted with hot water for 4 to 30 hours at a temperature condition of 70 to 90°C, a cosmetic composition for improving skin barrier function.
The method of claim 1,
The cosmetic composition has one or more uses selected from the group consisting of skin moisturizing enhancement, skin trouble prevention, and skin aging prevention, a cosmetic composition for improving skin barrier function.
delete delete delete The method of claim 1,
The fermented Schisandra chinensis extract is a cosmetic composition for improving skin barrier function, which has a liver and gastrointestinal protection effect.
Health functional food composition for improving skin barrier function, containing fermented Schisandra chinensis extract as an active ingredient.
The method of claim 7,
The fermented Schisandra chinensis extract is a health functional food composition for improving skin barrier function, which has a liver and gastrointestinal protection effect.
A quasi-drug composition for improving skin barrier function, containing fermented Schisandra chinensis extract as an active ingredient.
The method of claim 9,
The Schisandra chinensis fermentation extract is a quasi-drug composition for improving skin barrier function, which has a liver and gastrointestinal protection effect.
Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus paracasei tolerance and Lactobacillus paracasei sub. Inoculating one or more strains selected from the group consisting of Lactobacillus harbinensis with a number of bacteria of 2.0 to 7.0 at an OD600 absorbance value, and fermenting for 24 to 72 hours at 25 to 37°C, Manufacturing method of fermented Schisandra chinensis extract for improving skin barrier function.
delete

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