KR20200085126A - A cosmetic composition for preventing or improving skin wrinkles containing an extract of fermented omija as an effective ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing or ameliorating skin wrinkles containing a Schisandra chinensis fruit fermented extract as an active ingredient. Specifically, the present invention provides a cosmetic composition for preventing or ameliorating skin wrinkles containing a Schisandra chinensis fruit fermented extract as an active ingredient a health functional food composition and a pharmaceutical composition. The cosmetic composition, health functional food composition, and pharmaceutical composition of the present invention have low cytotoxicity, inhibit oxidation and inflammation of the skin, and have an effect of preventing or ameliorating skin wrinkles.

Description

A cosmetic composition for preventing or improving skin wrinkles containing Omija fermentation extract and it as an active ingredient

The present invention relates to a cosmetic composition for preventing or improving skin wrinkles containing Omija fermentation extract and it as an active ingredient.

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

Skin hydration is determined by the equilibrium between water transfer by osmotic pressure from the epidermis to the dermis and water loss in the epidermis by evaporation. Clinically, dry skin has a dry skin surface, poor elasticity, and is easily prone to scaling and rough skin. In addition, itchy symptoms may appear, and in some cases, symptoms such as dryness and cracking may accompany it. The level of moisturization of the epidermis is highly dependent on keratinocyte lipids, corneodesmosom, and natural moisturizing factor (NMF).

Among the skin layers, the dermis mainly consists of 　collagen and elastin fibers, which serve to support the skin. Therefore, when a problem occurs in the dermis, wrinkles are formed and skin elasticity disappears, so skin aging progresses. In particular, collagen is known to play the most important role in skin regeneration, skin moisture content, wound healing and wrinkle improvement, and is produced from fibroblasts. Specifically, collagen has a function to contain a large amount of moisture, and thus serves to supply moisture to the dermis. As it ages, the collagen's water-containing function decreases and wrinkles increase. Collagen also acts as a wound-healing agent by filling the wound area with continuous "collagen" production of fibroblasts when a wound is formed. On the other hand, the main component of connective tissue in the dermis of the skin that occupies most of the skin volume is known as “type 1 collagen”, and the “type 1 collagen” is known to be produced by fibroblasts (J. Clin. Immunol. 2005 , 25, 592).

The present invention is to provide a cosmetic composition, a health functional food composition and a pharmaceutical composition that can prevent or improve skin wrinkles by increasing the expression of the first type collagen using fermented extract derived from natural material Omija Did.

An object of the present invention is to provide a cosmetic composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

Another object of the present invention is to provide a method of manufacturing a fermented extract of Omija for preventing or improving skin wrinkles.

In order to achieve the above object, the present invention provides a cosmetic composition for preventing or improving skin wrinkles, containing Omija fermentation 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 preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a pharmaceutical composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

In order to achieve another object of the present invention, the present invention is a Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri in Omija extract, Including and inoculating and fermenting at least one strain selected from the group consisting of Lactobacillus paracasei sub.Tolerans and Lactobacillus harbinensis, Omija for preventing or improving skin wrinkles It provides a method for producing fermented extract.

According to the present invention, a cosmetic composition for preventing or improving skin wrinkles containing fermented extract of Omija as an active ingredient, a health functional food composition and a pharmaceutical composition have low cytotoxicity, inhibit oxidation and inflammation of the skin, and prevent skin wrinkles Or it has the effect of improving.

1 is a graph showing cell viability (A) and NO production (B) when treated with Omija fermentation extract on RAW264.7 cells.
Figure 2 is a graph showing the mRNA (C) expression of inflammatory cytokines IL-6(A), IL-β(B) and TNF-α when treated with Omija fermentation extract on RAW264.7 cells .
FIG. 3 shows that the reduction of expression of type 1 collagen and mRNA is inhibited when the Omija fermentation extract is treated with UVF irradiated HDF (human dermal cells).
Figure 4 shows the effect of inhibiting reactive oxygen species (ROS) when treated with Omija fermentation extract UVB-irradiated HDF cells.
Figure 5 shows the effect of procollagen (procollagen) biosynthesis when treated with Omija fermentation extract UVF irradiated HDF cells.
Figure 6 shows the increase in ERK-1/2 protein phosphorylation (activation), c-Fos protein and mRNA expression in HDF cells irradiated with Omija fermentation extract UVB 60mJ/cm ² , 4h.
FIG. 7 shows the results of hepatotoxicity test by hepatocellular death and necrosis caused by hepatotoxicity, and the results of experiments when the Omija fermentation extract was treated.
FIG. 8 shows the results of hepatotoxicity experiments due to liver size reduction caused by hepatocellular death and necrosis caused by hepatotoxic drugs and observation results based on GFP fluorescence when treated with Omija fermentation extract.
FIG. 9 shows the results of quantitatively measuring protein outflow through the intestine by measuring the fluorescence emitted after causing intestinal damage of zebrafish, and experimental results (absorbance) when treating the fermented extract of Omija.
FIG. 10 shows the results of quantitatively measuring protein outflow through the intestine by measuring the fluorescence emitted after causing intestinal damage of zebrafish, and the experimental results (% comparison) when the Omija fermentation extract was treated.

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

Among the skin layers, the dermis mainly consists of 　collagen and elastin fibers, which serve to support the skin. Therefore, when a problem occurs in the dermis, wrinkles are formed and skin elasticity disappears, so skin aging progresses. In particular, collagen is known to play the most important role in skin regeneration, skin moisture content, wound healing and wrinkle improvement, and is produced from fibroblasts. Specifically, collagen has a function to contain a large amount of moisture, and thus serves to supply moisture to the dermis. As it ages, the collagen's water-containing function decreases and wrinkles increase. Collagen also acts as a wound-healing agent by filling the wound area with continuous "collagen" production of fibroblasts when a wound is formed. On the other hand, the main component of connective tissue in the dermis of the skin that occupies most of the skin volume is known as “type 1 collagen”, and the “type 1 collagen” is known to be produced by fibroblasts (J. Clin. Immunol. 2005 , 25, 592).

On the other hand, the first collagen plays the most important role in skin regeneration, skin moisture content, wound healing and wrinkle improvement, and the Omija fermentation extract according to the present invention increases skin expression by increasing the expression of the first type collagen. It has the effect of preventing or improving.

Accordingly, according to an embodiment of the present invention, there is provided a cosmetic composition for preventing or improving skin wrinkles containing an Omija fermentation extract as an active ingredient.

In the present invention, Omija is a fruit that is opened in the vine deciduous tree of Omija family, and is called Omija because it has five kinds of sweet, bitter, spicy and salty taste. Omija is generally known to be effective in improving blood flow, preventing high blood pressure, stroke, cardiovascular disease, improving immunity, preventing diabetes, improving liver function, restoring respiratory disease, and improving respiratory disease.

In the present invention, the Omija fermentation extract may be obtained by extraction and fermentation from various organs or parts of Omija, and for example, may be a fermentation product of Omija hot water extract obtained by fermenting an extract obtained by hot water extraction of Omija Chung by-products. .

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

On the other hand, the cosmetic composition according to an embodiment of the present invention is not limited to being prepared in any formulation that is conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, 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. More specifically, it can be applied to produce functional cosmetics, which are basic cosmetics such as lotion, nourishing cream, moisture cream, essence, eye cream, lotion, pack, and essence.

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

Meanwhile, according to an embodiment of the present invention, the Omija fermentation 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 preventing and improving skin wrinkles within the weight% range. If less than the weight percent range, there may be a slight problem in preventing and improving skin wrinkles.

On the other hand, according to an embodiment of the present invention, it provides a health functional food composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

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

Meanwhile, the food composition may include ingredients commonly used in the health functional food composition in addition to the fermentation extract, for example, nutritional supplements, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, and organic acids , A protective colloidal emulsifier, a pH adjusting agent, a stabilizer, a preservative, glycerin, alcohol, a carbonizing agent used in carbonated beverages, or a combination thereof.

On the other hand, according to an embodiment of the present invention, to provide a pharmaceutical composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.

The pharmaceutical composition is a formulation such as external preparations for skin, granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions and emulsions using a pharmaceutically acceptable carrier and adjuvant, etc. It may be manufactured, but is not limited thereto.

In one example, the pharmaceutical composition may be formulated in a formulation such as an external preparation for skin, and in addition to the fermentation composition, for example, an aqueous component, an oil component, a powder component, alcohols, a moisturizing agent, an emulsifying agent, an ultraviolet absorber, a whitening agent, a preservative, an antioxidant , Surfactants, fragrances, colorants, various skin nutrients, etc. may be further appropriately included as necessary.

On the other hand, according to an embodiment of the present invention, Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus buchneri , Omija extract Preparation of an anti-wrinkle fermentation extract for preventing or improving skin wrinkles, comprising inoculating and fermenting at least one strain selected from the group consisting of Lactobacillus paracasei sub.Tolerans and Lactobacillus harbinensis Provide a method.

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

On the other hand, the fermentation may be carried out for 24 to 72 hours under the conditions of 25 to 37 ℃, inoculating microorganisms in the bacterial count of 2.0 to 7.0 at the OD 600 absorbance value. Specifically, when fermentation is performed with less than 2.0 bacteria at OD 600 absorbance, there is a problem that bacteria may not grow, and when fermentation is performed with more than 7.0 bacteria 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, fermentation may be difficult to be effectively performed, and when fermentation exceeds 72 hours, contamination may be a problem.

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

Example

Example 1. Preparation of Omija fermentation extract

100 g of the remaining Omija cheong by-product used in the production of Omija drink was prepared, and then the Omija by-product was mixed with 1000 g of water, followed by hot water extraction at 80° C. for 24 hours to obtain an Omija hot water extract.

A strain of Lactobacillus paracasei sub.Tolerans was inoculated to the hot water extract to obtain an Omija fermentation extract. During the fermentation, the number of bacteria was inoculated with an absorbance value of OD 600 from 4.0 to 6.0, and byproducts of Omija were prepared at a ratio of 10 wt%, and inoculated with 1 to 2% of bacteria (OD 600 absorbance value 0.1) in the culture solution, and incubated 48 Incubated for hours. The culture solution was centrifuged for 30 minutes to collect the supernatant free from bacteria, and the collected supernatant was filtered with a filter (5 μm pore size) to finally obtain an Omija fermentation extract.

Example 2. Omija fermentation extract 30% ethanol extract

The fermented extract of Omija prepared according to Example 1 was extracted using a PB-600 adsorption resin to obtain a 30% ethanol extract of Omija fermentation extract. At the time of extraction, 300 g of PB-600 adsorption resin is prepared, and 1000 ml of Omija fermentation extract is flowed. Then, 500 ml of DW, 10% ethanol, and 30% ethanol were sequentially flowed. Finally, 30% ethanol extract was obtained from Omija fermentation extract that had been shed with 30% ethanol.

[Experiment 1. Cytotoxicity test of Omija fermentation extract]

The macrophage cell line RAW264.7 is pre-sold from the Korea Cell Line Bank (KCLB, Seoul, Korea) and 5% CO using DMEM medium (Welgene, Gyeongsan, Korea) containing 10% FBS and 1% antibiotic (penicillin/streptomycin). _It was used after subcultured 2-3 times in a 37°C incubator where ₂ was present.

Cell viability measurement

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

NO(Nitric Oxide) measurement

In order to measure the amount of NO generated in the cells, 30% ethanol extract of Omija fermentation extract of Example 2 was treated on Raw 264.7 cells, stimulated with 1 ug/mL of LPS, and Griess in an amount equivalent to the cell culture after 24 hours of incubation. After adding a reagent (1% sulfanilamine, 0.1% naphthylethylene diamine di-hydrochloride, 2% phosphoric acid) and reacting, absorbance was measured at 540 nm. The concentration of NO was calculated based on the quantitative curve of the NaNO ₂ standard solution (see FIG. 1 (B)).

Cytokine measurement

Raw 264.7 cells were treated with 30% ethanol extract of the Omija fermentation extract of Example 2 and LPS 1 ug/ to measure the expression level of the mRNA of IL-6, IL-1β and TNF-α, which are inflammatory cytokines produced in the cells. After stimulation with mL and cultured for 24 hours, the culture supernatant was obtained. Cytokine concentrations were measured using an enzyme-linked immunosorbent assay (ELISA) kit. Each 96-well plate was coated with 100 μL of capture antibody for each cytokine, and then left at 4° C. for 12 hours. The plate was washed with PBS (phosphate-buffered saline) containing 0.05% tween-20™ and 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, and the detection antibody and the enzyme reaction solution (SAv-HRP reagent) were added, followed by the reaction by adding the substrate ABTS solution. The color 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 RAW264.7 macrophage cell line according to the content of 30% ethanol extract of Omija fermentation extract of Example 2, as the content of Omija fermentation extract at the same concentration increased, NO production decreased. appear. Therefore, it was found that the anti-inflammatory effect increased in proportion to the concentration of the fermented extract of Omija. On the other hand, the NO production inhibitory effect of the Omija fermentation extract was confirmed to exhibit 87% NO production inhibitory effect at a maximum concentration of 0.5 mg/ml without cytotoxicity (see FIG. 1).

On the other hand, it was confirmed that the expression levels of the inflammatory cytokines IL-6, IL-1β and TNF-α mRNA decreased significantly with increasing concentrations of Omija fermentation extract (see FIG. 2).

[Experiment 2. Experiment for promoting skin wrinkle improvement factor protein and mRNA expression of Omija fermentation extract]

Measurement of cell proliferation rate and viability

Cytotoxicity test was performed by the cell count and MTS assay method, specifically, the concentration of the 30% ethanol extract of Omija fermentation extract of Example 2 was treated with 25, 50, and 100 ug/ml concentration, respectively, and confirmed by Cell Counter. (See Figures 3 (A) and (B)).

Western blot experiment

In order to confirm the expression of type 1 collagen in HDF cells irradiated with UVB, it was confirmed by Western blotting.

Specifically, after 24 hours of inoculating human dermal cell HDF, 30% ethanol extract of the Omija fermentation extract of Example 2 was added and EGF was added as a positive control, respectively, and cultured in DMEM medium, 37°C, 5% CO ₂ . After incubation for 24 hours, cells were recovered and crushed to extract only proteins and stored at -40°C. The extracted protein was quantified through BCA assay. After diluting the protein amount of each sample to 30g, electrophoresis was performed on a 12% SDS-gel and transferred to a PVDF membrane. After reacting with the primary antibody (rabbit anti-aquaporins-3 antibody), the remaining antibody was washed and removed to react with the 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 generate a color reaction was added. The colored band was measured by image analysis (BioRad, Universal Hood, USA) (see FIG. 3).

Measurement of inhibitory effects on reactive oxygen species

To analyze the intracellular ROS level, a cell-permeable non-fluorescent probe, DCFH-DA (2',7'-dichlorofluorescin diacetate) (Sigma-Aldrich Co., St. Louis, MO) was used. After incubating for 12 hours under the administration of 30% ethanol extract of 100 μg/ml of Omija fermentation extract of Example 2, UVB-treated HDF cells were cultured in 10 μM DCFH-DA at 37° C. for 30 minutes. The cells were tested using a fluorescence microscope (Nikon Eclipse Ti inverted fluorescence microscopy) (Nikon, Tokyo, Japan). To measure the fluorescence intensity, cells were washed with PBS and trypsinized to remove them. Dropped cells were suspended in PBS containing 1% FBS after washing. The fluorescence intensity of each sample is measured at 485 nm excitation wavelengths and 525 nm emission wavelengths using a fluorescence microplate reader (SpectraMax ^® M4, Molecular Devices, CA), and the corresponding protein concentration. Standardized to

As a result, 　UVB treatment increased the 　ROS　 level in the cells by 3 to 4 times, which was inhibited by 30% ethanol extract of the Omija fermentation extract of Example 2. It is determined that the cell protective effect of the Omija fermentation extract against oxidative stress induced by UVB is the result of the antioxidant activity of the Omija fermentation extract (see FIG. 4 ).

Measures the effect of promoting procollagen biosynthesis

After adding 2 ml of DMEM culture solution to a 40 mm cell culture dish, HDF cells were inoculated at a concentration of about 1.2x10 ⁵ , and cultured for 24 hours in a 37°C, 5% CO ₂ environment. Thereafter, UVB was irradiated under conditions of 60 mJ/cm ² and 4 h, and then cultured for 3 days by replacing the medium with 100 μg/ml of 30% ethanol extract of the Omija fermentation extract of Example 2. Then, the culture medium was harvested and centrifuged for 5 minutes at 4°C and 7500 rpm for 5 minutes to change the protein expression level of the type 1 procollagen (Procollagen Type IC Peptide EIA Kit, Takara, Shiga, Japan) using the ELISA method. It was confirmed (see Fig. 5).

RT-PCR experiment

First, human dermal fibroblasts (HDF) cells were cultured in DMEM medium, 37° C., 5%, CO ₂ by adding 30% ethanol extract of Omija fermentation extract of Example 2 to human dermal fibroblasts (HDF) cells. After incubation for 24 hours, cells were recovered and RNA was extracted to perform PCR using reverse transcriptase and cDNA (complementary DNA). GAPDH was used as a standard. The band expressed through electrophoresis of agarose gel was measured by image analysis (BioRad, Universal Hood, USA) (see FIG. 6).

Experiment result

In the case of cell proliferation rate and cell viability according to the concentration of the 30% ethanol extract of Omija fermentation extract of Example 2, it was confirmed that the concentration was increased in a dependent manner. On the other hand, it was confirmed that the type 1 collagen and mRNA expression reduction is suppressed under the condition irradiated with UVB (see FIG. 3).

On the other hand, UVB treatment increases the intracellular “ROS” level by 3 to 4 fold, and it was confirmed that the Omija fermentation extract of Example 1 inhibits the intracellular ROS level. The cell protective effect of the Omija fermentation extract against oxidative stress induced by UVB is determined to be a result of the antioxidant activity of the 30% ethanol extract of the Omija fermentation extract of Example 2 (see FIG. 4 ).

On the other hand, it was confirmed that procollagen biosynthesis of type 1 collagen is promoted when treating 30% ethanol extract of Omija fermentation extract of Example 2 in UVB irradiated HDF cells (see FIG. 5).

In addition, when treating the Omija ferment extract of Example 1 at the time of UVB irradiation at a concentration of 100 ug/ml, ERK-1/2 p38 MAPK, JNK-1 in HDF cells specifically irradiated with UVB 60 mJ/cm ² , 4h /2, c-JUN protein phosphorylation (activation) increase and c-Fos and c-JUN protein and mRNA expression was confirmed to cause an increase (see FIG. 6).

[Experiment 3. Liver protection and gastrointestinal tract protection experiment of Omija fermentation extract]

Liver protection experiment

Based on the hepatic cell death caused by the hepatotoxic drug and the reduction in the size of the liver caused by necrosis, based on standard data of liver size change by the hepatotoxic drug acetaminophen, the Omija fermentation extract of Example 2 The pre-treatment of the 30% ethanol extract followed by treatment with acetaminophen was intended to confirm the hepatoprotective effect of the 30% ethanol extract of the Omija fermentation extract of Example 2.

Specifically, the adult zebrafish used for the test used a healthy strain of AB and an LFABP-GFP isoform transformant established through genetic manipulation.

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

Control group: No treatment group

Experimental group 1: acetaminophen treated group

Experimental Groups 2-4: Acetaminophen treated group after pretreatment with 30% ethanol extract of Omija fermentation extract of Example 2

Specifically, the concentration of the drug to be treated for each well to be tested was entered individually, and on the second day, 30% ethanol extract of the Omija fermentation extract of Example 2 was added according to the concentration, and the drug added by carefully rotating the well plate circularly Mix evenly. Next, transferred to an incubator, incubated for 24 hours, and then acetaminophen was added on the third day. On the 4th and 5th day, the size and shape of the liver were measured by fluorescence microscopy, and mean±SD results were obtained by averaging data from 10 animals (see FIGS. 7 and 8 ).

Gastrointestinal tract protection experiment

KR68766 is a fluorescent substance that binds with albumin and attempts to quantitatively measure protein outflow through the intestine by measuring the fluorescence flowing out of the medium when intestinal damage occurs after staining zebrafish embryos.

Specifically, zebrafish adults were reared at 28.5°C with contrast of 14 hours during the day and 10 hours at night. A couple of male and female were placed in a dedicated mating cage on the afternoon of the previous day, and the fertilized eggs were collected the next morning. The collected fertilized eggs are transferred to a petri dish and generated in an incubator at 28.5℃, and the process of development is observed through a dissecting microscope.

On the 24 well plate, 15 embryos per well were placed on the third day after fertilization. After being divided into the experimental group and the control group, the experimental group was pretreated with 30% ethanol extract of Omija fermentation extract of Example 2 for 24 hours. The next day, after dyeing with KR68766, indomethacin was treated for 24 hours to induce intestinal damage. The next day, the medium was taken to measure fluorescence absorbance (see FIGS. 9 and 10).

Experiment result

When the 30% ethanol extract of the Omija fermentation extract of Example 2 was treated, it was confirmed that it showed a protective effect of liver damage by acetaminophen at a concentration of 10 ug/ml of the drug, and that the liver protective effect increased as the concentration increased. I could confirm. However, liver size decreased in the 100 ug/ml treatment group, and 100 ug/ml + acetaminophen group showed lethality (see FIGS. 7 and 8).

In the case of the gastrointestinal tract protective effect, the protective effect increased as the drug concentration increased, similar to the hepatotoxicity test, but was not significantly different at the concentration of 1 ug/ml, and the drug toxicity was seen in the 100 ug/ml treatment group, and 100 ug/ml + Indomethacin group all showed lethality (see FIGS. 9 and 10).

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

Formulation Example 1. Flexible Cosmetics

The formulation example of the softening makeup was prepared as in the following Table 1 according to a conventional method.

Formulation Example 2. Essence

The formulation example of the essence was prepared as shown in Table 2 below according to a conventional method.

Formulation Example 3. Pack

The formulation example of the pack was prepared as in Table 3 according to a conventional method.

According to the experimental examples of the present invention, in the case of the Omija fermentation extract of the present invention according to the embodiments, the effect of preventing or improving skin wrinkles by increasing the expression of the first type1 collagen known as a factor related to skin wrinkles It was confirmed that it has a low cytotoxicity, and thus it can be used as a cosmetic raw material by suppressing oxidation and inflammation of the skin and preventing or improving skin wrinkles.

Claims (11)

A cosmetic composition for preventing or improving skin wrinkles, containing Omija fermentation extract as an active ingredient.
According to claim 1,
The Omija fermentation extract is a fermentation product of Omija hot water extract, a cosmetic composition for preventing or improving skin wrinkles.
According to claim 1,
The Omija fermentation extract is Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus paracasei Tolerance para. ) And Lactobacillus harbinensis (Lactobacillus harbinensis) is a fermented with one or more strains selected from the group consisting of a cosmetic composition for preventing or improving skin wrinkles.
According to claim 1,
The Omija fermentation extract is contained in 0.1 to 5% by weight relative to the total weight of the cosmetic composition, cosmetic composition for preventing or improving skin wrinkles.
According to claim 1,
The Omija fermentation extract is to have a protective effect on the liver and gastrointestinal tract, cosmetic composition for preventing or improving skin wrinkles.
A health functional food composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.
The method of claim 6,
The Omija fermentation extract is to have a protective effect on the liver and gastrointestinal tract, a health functional food composition for preventing or improving skin wrinkles.
A pharmaceutical composition for preventing or improving skin wrinkles containing Omija fermentation extract as an active ingredient.
The method of claim 8,
The Omija fermentation extract is to have a protective effect on the liver and gastrointestinal tract, pharmaceutical composition for preventing or improving skin wrinkles.
To the extract of Omija , Pichia kudriavzevii, Saccharomycopsis fibuligera, Lactobacillus buchneri, Lactobacillus paracasei Tolerance and Lactobacillus paracasei subcasei Including the step of inoculating and fermenting one or more strains selected from the group consisting of Lactobacillus harbinensis (Lactobacillus harbinensis) , a method for manufacturing a fermented extract of Omija for skin wrinkle prevention or improvement.
The method of claim 10,
The fermentation is performed for 24 to 72 hours, a method of manufacturing a fermented extract of Omija for preventing or improving wrinkles.

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