KR20210146647A - Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing skin aging or alleviating wrinkles comprising a Weissella cibaria cell lysate. The composition comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, or a cytolysate of an extract or culture thereof of the present invention has no cytotoxicity, inhibits collagenase activity, and exhibits an effect of inhibiting the production of MMPs increased by fine dust and a reduced effect of promoting the synthesis of type 1 collagen (COL1A1), thereby being useful as a cosmetic, health functional food or health food composition for preventing skin aging or alleviating wrinkles.

Description

Cosmetic composition for skin anti-aging or anti-wrinkle containing Weissella cibaria cell lysates}

The present invention relates to a cosmetic composition for anti-aging or wrinkle improvement comprising a Wasella civaria cell lysate, and more particularly, to a cosmetic composition for preventing skin aging or wrinkle improvement comprising a Wasella civaria cell lysate. it's about

Impressions are the appearance that others see, and are formed by clothes, skin, facial expressions, makeup, posture, etc. In modern society, impressions are very important because the inside of a person is greatly evaluated indirectly through impressions, and more and more people express themselves through their appearance. As the importance of appearance increases, the demand for cosmetics is increasing significantly. In particular, among cosmetics, interest and demand for wrinkle-improving functional cosmetics that also have the characteristics of pharmaceuticals are steadily increasing.

Recently, consumers are not only interested in the efficacy of finished cosmetics, but are also increasingly interested in the efficacy and stability of each raw material used in cosmetics. Therefore, research on the development of functional raw materials of natural materials harmless to the human body is being actively conducted in many cosmetic research institutes, and most recent cosmetics are made using materials with guaranteed stability or active ingredients. In particular, among many functional cosmetics, the development of raw materials for wrinkle-improving functional cosmetics is still in progress, and the launch rate of cosmetics using new and various functional materials is also increasing. In recent years, various types of cosmetics that have the effect of protecting the skin from harmful environmental substances such as fine dust have been released, and research and development of materials and formulations for this purpose are being actively conducted. When all of these are taken together, the need for research and development of natural-derived materials with guaranteed stability to show anti-aging effects from harmful environmental substances such as fine dust is emerging.

With the rapid development of modern society, the problem of environmental pollution is also accompanied. Among air pollutants, particulate matter (PM) has a particle size of 10 μm, and _{ionic components such as nitrate (NO 3} ^- ), ammonium (NH ₄ ⁺ ), and sulfate (SO ₄ ^2- ) are generated by combustion. It is made up of metals, carbon compounds, and metal compounds, causing various diseases such as respiratory diseases, cardiovascular diseases, and skin diseases, which adversely affect human health, which is a social problem. The first place that fine dust comes into contact with the human body is the skin. Fine dust damages the human epidermal barrier function, and various contaminants penetrate into the skin through the damaged skin barrier, resulting in acne, pigmentation, and aging dermatitis. Even worse, it can lead to skin diseases including hair loss and skin cancer.

As the mechanism that causes skin aging, ROS (Reactive oxygen species) increases as oxidative stress occurs by fine dust, UV or external stimuli, the MAPK (Mitogen-activated protein kinase) mechanism is activated, and AP-1 (Transcription factor activator) It has been reported that the expression and activity of matrix metalloproteinase (MMPs) that degrade collagen by affecting protein 1) or NF-kB (nuclear factor-kB) substances increase, resulting in lowered collagen synthesis, resulting in skin aging. Fine dust, one of the factors that causes skin wrinkles, is very small and can easily penetrate the skin, destroying the skin barrier, pollutants in the fine dust affect the skin immune system, and collagen and elastin to maintain skin elasticity It destroys the skin, resulting in wrinkles and damage to the skin. MMPs, whose expression and activity are increased in skin epithelial and dermal cells by external factors including fine dust, can be largely classified into Collagenases, Gelatinases, and Stromelysins according to their structure and functional characteristics. Collagenases include MMP-1, -8, -13, and -18, which have a role in cutting collagen structures. Glatinases include MMP-2 and MMP-9, which modify and degrade the structure of basement membrane collagen. MMP-3, -10, -11, and -19 are Stromelysins, which degrade Proteoglycans, Martix glycoproteins, and collagen, and activate various MMPs to play an important role in wrinkle formation. MMPs increased by such fine dust are expressed in epithelial cells, human Keratinocytes and dermal cells, human dermal fibroblasts, etc. The elasticity is reduced and the skin wrinkles are formed. In the dermal layer of the skin, type 1 collagen (COL1A1) occupies 80 to 90%, and other types of collagen and fibronectin are also composed. Due to various external factors and age increase, the activity of collagenase increases, decomposing collagen, and thus the smoothness, moisture, and elasticity of the skin are reduced. For this reason, reduction in collagenase activity and collagen gene and increase in MMPs expression are major factors in wrinkle formation. .

Lactic acid bacteria are important microorganisms that are essential for the manufacture of various fermented foods such as yogurt, cheese and fermented milk. Lactic acid bacteria not only ferment food, but also enhance human immunity, prevent food poisoning, prevent constipation, and have intestinal effects, so it has been developed through research and development for a long time. Recently, it has been reported to be effective in preventing cancer. Because of the tremendous efficacy of these lactic acid bacteria, interest in various lactic acid bacteria fermented products and foods is increasing year by year. Although the microorganisms of kimchi are not fully identified, the microorganisms found in kimchi are largely divided into three genera: Lactobacillus, Weissella , and Leuconostoc. Among them, Weissella cibaria , a representative microorganism of the genus Weissella, is a Gram-positive, non-motile short rod, Catalase-negative, facultative anaerobic bacteria that grow well in both aerobic and anaerobic conditions. have a characteristic In addition, Wasella is known to be abundant in fresh kimchi, and has been found to have anticancer, antiviral, anti-obesity, immune enhancement, and atopy improvement effects, but research as a cosmetic material is still insignificant.

Therefore, the present inventors confirmed the effect of preventing skin aging and improving skin wrinkles due to fine dust by inhibiting the activity of the Weissella civaria cell lysate and regulating the expression of genes related to wrinkle generation due to fine dust, and was completed.

Korean Patent No. 10-1742029

It is an object of the present invention to provide a cosmetic composition for improving skin wrinkles or enhancing elasticity.

Another object of the present invention is to provide a cosmetic composition for preventing or improving skin aging.

Another object of the present invention is to provide a health functional food composition for improving skin wrinkles or enhancing elasticity.

Another object of the present invention is to provide a health functional food composition for preventing or improving skin aging.

Another object of the present invention is to provide a health food composition for improving skin wrinkles or enhancing elasticity.

Another object of the present invention is to provide a health food composition for preventing or improving skin aging.

In order to achieve the above object,

The present invention provides a cosmetic composition for improving skin wrinkles or enhancing elasticity, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

In addition, the present invention provides a cosmetic composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

Furthermore, the present invention provides a health functional food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

Furthermore, the present invention provides a health functional food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

In addition, the present invention provides a health food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

Furthermore, the present invention provides a health food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

The composition comprising the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is not cytotoxic, and inhibits collagenase activity, Since it exhibits an increased inhibitory effect on the production of MMPs and a reduced effect on the synthesis of collagen type 1 (COL1A1), it may be useful as a cosmetic, health functional food or health food composition for skin aging prevention and wrinkle improvement.

1 is a result of confirming the cytotoxicity of the Weissella cell lysate of the present invention in human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1).
Figure 2 is the result of confirming the activity inhibitory effect of the collagen degrading enzyme of the Weissella cell lysate of the present invention.
3 is a result confirming the inhibitory efficacy of MMPs expression increased by fine dust in the Weissella cell lysate of the present invention in human keratinocytes (HaCaT).
4 is a result confirming the effect of increasing the expression of collagen type 1 (COL1A1) reduced by fine dust in the Weissella cell lysate of the present invention in human fibroblasts (CCD-986Sk) and human dermal fibroblasts (HDF-1) .

Hereinafter, the present invention will be described in detail.

Cosmetic composition for improving skin wrinkles or enhancing elasticity

The present invention provides a cosmetic composition for improving skin wrinkles or enhancing elasticity, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.

In the cosmetic composition of the present invention, the skin wrinkles may be caused by fine dust.

Aging also involves an enzyme called Matrix Metalloproteinase (MMP), a collagen degrading enzyme. That is, the synthesis and decomposition of extracellular matrix such as collagen in vivo is properly regulated. As aging progresses, collagen synthesis decreases, and expression of matrix metalloproteinase (MMP), an enzyme that decomposes collagen, is promoted. As a result, the elasticity of the skin decreases and wrinkles are formed.

Therefore, in the case of a substance capable of regulating or inhibiting MMP expression induced in activation in cells, it can be used as a material for functional cosmetics for improving skin elasticity, improving wrinkles, and inhibiting aging.

There are 23 types of matrix metalloproteinase (MMP) that catalyze collagen degradation in mammals. Among them, MMP types increased by UV light are known as 1, 3, and 9, and these three types of MMPs are collagen type. It is an enzyme that breaks down 1 and 3. Specifically, in the case of MMP-1, the middle of the collagen fibers are cut, and MMP-3 and MMP-9 serve to subdivide and cut the cut collagen fibers.

That is, the composition comprising the Weissella cibaria strain, its culture, its lysate, its extract, or cell lysate of the culture according to the present invention significantly reduces the expression of the MMPs gene, so that the Decomposition can be inhibited, and by increasing the synthesis of collagen in the skin tissue, skin elasticity improvement, wrinkle improvement, and aging can be suppressed (see Experimental Example 3).

The cosmetic composition according to 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, pack, It may be one or more formulations selected from the group consisting of soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, and may also be used in the form of external preparations for skin. Components other than the above essential components can be appropriately selected and formulated by those skilled in the art without difficulty according to other formulations or purposes of use.

The cosmetic composition of the present invention preferably contains a cosmetically acceptable medium or base. It is in any dosage form suitable for topical application, for example solutions, gels, solid or kneaded dry products, emulsions obtained by dispersing the oily phase in an aqueous phase, suspension microemulsions, microcapsules, microgranules or ionic (liposomes) and/or bi It may be provided in the form of a warm, vesicular dispersion, or in the form of a cream, toner, lotion, powder, ointment, spray or cone stick. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant. In addition, the cosmetic composition of the present invention can be prepared in the form of an adjuvant that can be applied topically or systemically commonly used in the field of dermatology by containing a dermatologically acceptable medium or base, and these compositions are conventional in the art. It can be prepared according to the method.

In addition, the cosmetic composition includes a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, and water in addition to the extract of the present invention. , ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the cosmetic field, such as any other ingredients. These adjuvants are introduced in amounts generally used in the field of cosmetology.

Cosmetic composition for preventing or improving skin aging

The present invention provides a cosmetic composition for preventing or improving skin aging, comprising a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.

In the cosmetic composition of the present invention, the skin wrinkles may be caused by fine dust.

The cosmetic composition according to 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, pack, It may be one or more formulations selected from the group consisting of soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, and may also be used in the form of external preparations for skin. Components other than the above essential components can be appropriately selected and formulated by those skilled in the art without difficulty according to other formulations or purposes of use.

Health functional food or health food composition for skin wrinkle improvement or elasticity enhancement

The present invention provides a health functional food or health food composition for improving skin wrinkles or enhancing elasticity, including a Weissella cibaria strain, a culture thereof, a lysate thereof, an extract thereof, or a cell lysate of the culture.

In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.

In the composition of the present invention, the skin wrinkles may be caused by fine dust.

When the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is used as a health functional food and health food composition, the type of food is not particularly limited. Examples of health functional food to which the Weissella cibaria strain of the present invention, its culture, its lysate, its extract, or cell lysate of the culture can be added may be in the form of tablets, capsules, pills or liquids. Examples of health food include various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes, etc. of health functional food and health food composition.

Weissella cibaria strain according to the present invention, its culture, its lysate, its extract or health functional food and health food composition containing a cell lysate of the culture is added to food as it is, or other food or food ingredients can be used together, and can be appropriately used according to a conventional method. Weissella cibaria strain, its culture, its lysate, its extract, or the mixed amount of the cell lysate of the culture may be suitably determined depending on the purpose of its use (for prevention or improvement). In general, the amount of the composition in the health functional food and health food composition may be added in an amount of 0.1 to 90 parts by weight based on the total weight of the food. However, in the case of long-term intake for health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the Weissella cibaria strain, its culture, A lysate thereof, an extract thereof, or a cell lysate of a culture may be used in an amount greater than or equal to the above range.

The health functional food and health food composition of the present invention is an essential ingredient in the indicated ratio, and the present invention Weissella cibaria strain, its culture, its lysate, its extract or cell lysate of the culture. The other ingredients are not particularly limited and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 health functional food and health food composition of the present invention.

In addition to the above, the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or health functional food and health food composition containing a cell lysate of the culture are various nutrients, vitamins, minerals (electrolytes) ), synthetic and natural flavoring agents, colorants and thickeners (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives , glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. In addition, the health functional food and health food composition of the present invention may contain natural fruit juice, fruit juice for the production of fruit juice drinks, and vegetable drinks.

These components may be used independently or in combination. Although the ratio of these additives is not so important, the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or 100 parts by weight of a health food composition containing a cell lysate of the extract or culture It is generally selected in the range of 0.1 to about 20 parts by weight of sugar.

Health functional food or health food composition for preventing or improving skin aging

The present invention provides a health functional food or health food composition for preventing or improving skin aging, comprising a Weissella cibaria strain, its culture, its lysate, its extract, or a cell lysate of the culture.

In one embodiment of the present invention, the composition comprises a strain, a culture thereof, a lysate thereof, an extract thereof, or a cell suspension of a culture, a cell homogenate, a cell lysate, a cell supernatant, a cell filtrate, a cell pellet, or one or more cells, or may include them. More preferably, it may be a cell lysate of Weissella cibaria strain culture.

In the composition of the present invention, the skin wrinkles may be caused by fine dust.

When the Weissella cibaria strain of the present invention, its culture, its lysate, its extract or cell lysate of the culture is used as a health functional food and health food composition, the type of food is not particularly limited. Examples of health functional food to which the Weissella cibaria strain of the present invention, its culture, its lysate, its extract, or cell lysate of the culture can be added may be in the form of tablets, capsules, pills or liquids. Examples of health food include various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes, etc. of health functional food and health food composition.

The present invention will be described in more detail through the following examples. However, the following examples are only intended to embody the contents of the present invention, and the present invention is not limited thereto.

<Example 1> Preparation of Weissella civaria cell lysate

Weissella civaria used in the present invention was isolated from commercially available kimchi. The kimchi was crushed finely, filtered, spread on MRS agar (Lactobacilli MRS agar), and cultured for 16 hours. After selecting several types of cultured colonies, they were further cultured in the MRS borth, and 16S ribosoma RNA sequence analysis was performed. Among the analyzed strains, Weissella civaria bacteria, whose efficacy in improving skin aging and wrinkle have not yet been identified, were selected and inoculated in MRS borth medium and cultured at 30° C. for 24 hours for use in this study.

After spreading Weissella civaria on MRS agar medium (Lactobacilli MRS agar), it was cultured at 30° C. for 16 hours to obtain a single colony. After the obtained single colony was inoculated in Lactobacilli MRS broth, incubated at 30° C. for 16 hours, the cells were centrifuged at 7,000 rpm for 10 minutes to remove the culture medium, and PBS (Phosphate-buffered Saline) buffer 3 The medium was washed twice to remove media components. Only the cells from which the medium component has been removed are obtained by centrifugation, and after dispensing into 1.5 ml tubes, 400 μl of PBS buffer is added to lyse the cells with a sonicator, and centrifuged at 13,000 rpm for 5 minutes. It was transferred and used after protein quantification. The Weissella civaria cell lysate used in the present invention was hereinafter referred to as the Weissella lysate.

<Preparation Example 1> Cell culture

Experimental human skin cells in human keratinocytes used to form the cell (HaCaT), human fibroblasts (CCD-986Sk), human dermal fibroblasts (HDF-1) have been incubated at 37 ℃, CO ₂ incubator for new DMEM every 3 days (Dulbecco Modified Eagle Medium) or after washing with PBS, Tyrpsin-EDTA solution was added to separate the cells from the culture dish and subcultured and maintained.

<Experimental Example 1> Confirmation of cytotoxicity

To determine the toxicity of Weissella cibaria cell lysates in human skin cells, human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1) were each in a 24-well-plate. Incubated for 24 hours in _{a 37 ℃ CO 2} incubator by dividing the same amount of cells. After culturing, the Wasella lysate was treated by concentration (0 μg/ml, 100 μg/ml, 200 μg/ml), and then _{further cultured in a 37° C. CO 2} incubator for 24 hours. After incubation, wash with PBS solution, add 100 μl of Trypsin-EDTA solution, incubate for 3 minutes in the incubator under the same conditions, completely remove the cells from the culture dish, add 400 μl of DMEM medium each, and pipette cells were mixed well. To count the cells, 10 μl of cells were loaded into a hemocytometer and the number of viable cells was counted.

As a result of confirming the cytotoxicity of Weissella lysates in human keratinocytes (HaCaT), human fibroblasts (CCD-986Sk), and human dermal fibroblasts (HDF-1), as shown in Table 1 and Figure 1, 100 There was no toxicity at the μg/ml concentration. In addition, when the Weissella lysate was treated at a concentration of 200 μg/ml, it was toxic to HaCaT cells, but not significantly toxic to the other two cells. In subsequent experiments, the Weissella lysate was used at a concentration of 100 μg/ml.

<Experimental Example 2> Confirmation of inhibition of collagenase activity of Wasella civaria cell lysate

To determine whether the Wasella lysate inhibits collagenase activity that degrades collagen, various concentrations (0, 40, 80, and 160 μg/ml) of the Wasella lysate and the substrate 0.6 mM FALGPA were placed in 96-well plates. (N-[3-(2-Furyl)acryloyl]-Leu-Gly-Pro-Ala) was reacted in 50mM Tricine buffer (pH 7.5) containing 400mM _{NaCl 2} and 10mM CaCl _{2 at room temperature for 15 minutes and then 0.2 units} /ml Collagenase was added, and absorbance was measured every minute at a wavelength of 345 nm to measure the degree to which the substrate was decomposed by collagenase. Here, the negative control group refers to a blank containing only the substrate and the buffer.

As a result, as shown in FIG. 2 , it was confirmed that the Weissella lysate decreased the activity of the collagenase in a concentration-dependent manner. Through the above experimental results, it was found that the Wasella lysate of the present invention had the effect of preventing skin aging and improving skin wrinkles by inhibiting the activity of collagen degrading enzyme.

<Experimental Example 3> Confirmation of expression of MMPs and collagen genes by fine dust of Weissella civaria cell lysate

In order to determine whether the Weissella lysate changes the expression of genes involved in skin aging in human skin cells, the cells were each divided into 6 well-plates with the same number of cells _{and cultured in a 37° C. CO 2} incubator for 24 hours. . After culturing, the Wasella lysate was treated with different concentrations (0, 50 and 100 μg/ml), and then _{pre-treated in a 37° C. CO 2} incubator for 1 hour, fine dust was added, and further cultured for 16 hours. In this experiment, fine dust of 4 μm or less (NIST2786), which is a standard reference material (SRM) of the National Institute of Standards and Technology (NIST), was treated for fine dust.

After additional incubation, remove the culture solution containing the sample, remove the medium components with PBS solution, add 1 ml of RNAiso Plus (Takara) solution, and then transfer to each tube and add 200 μl of chloroform solution to each tube. After addition, the mixture was sufficiently mixed using a vortex mixer and then reacted at room temperature for 10 minutes. Next, after centrifugation for 15 minutes at 13000 g, 4 ℃ condition, only the aqueous phase was mixed with 500 μl isopropanol (isopropanol) in a new tube, and then reacted at room temperature for 10 minutes. After centrifugation under the same conditions, the supernatant was removed, 75% ethanol was added, and centrifuged again under the same conditions, and the ethanol, the supernatant, was completely removed and dried. After adding 40 μl of DEPC-treated water to each tube, the RNA concentration was measured and cDNA was synthesized using the cDNA synthesis kit (TAKARA) at the same concentration. The synthesized cDNA was used for Reverse Transcription Polymerase Chain Reaction (RT-PCR) together with each primer, and the expression level of each gene was confirmed on 1% agarose gel.

As a result of confirming the effect of the Wasella lysate on the expression of MMPs increased by fine dust, as shown in FIG. 3 , fine dust increased the gene expression of MMP-3 and MMP-9 in human keratinocytes, but It was confirmed that the increased expression of MMP-3 and MMP-9 caused by fine dust was inhibited in the cells pretreated with the Wasella lysate.

MMPs increase the expression of collagen-degrading enzymes to break down collagen, thereby causing skin wrinkles. Therefore, as a result of confirming the effect of the Wasella lysate on the expression of the collagen gene reduced by fine dust, fine dust reduced the expression of the collagen gene in human fibroblasts and human dermal fibroblasts, as shown in FIG. 4 , In the cells pretreated with the Wasella lysate, it was confirmed that the decrease in the expression of collagen genes caused by fine dust was suppressed.

Therefore, through the above experimental results, the Wasella lysate inhibited the expression of MMPs increased by fine dust and inhibited the decomposition of collagen caused by the increased MMPs, thereby preventing skin aging and improving skin wrinkles. .

The above results suggest that the Wasella civaria lysate of the present invention has an effect of preventing skin aging and improving skin wrinkles.

So far, the present invention has been looked at with respect to preferred embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Preparation example of cosmetics

The Weissella civaria lysate according to the present invention can be prepared into various types of cosmetics depending on the purpose. The following exemplifies the manufacturing method of several cosmetics containing the Wasella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Cosmeceutical Preparation Example 1> Preparation of softened lotion

Weissella siberia lysate 10.0 parts by weight

1,3-butylene glycol 1.00 parts by weight

Disodium EDIT 0.05 parts by weight

allantoin 0.10 parts by weight

Dipotassium glycyrrhizate 0.05 parts by weight

citric acid 0.01 parts by weight

sodium citrate 0.02 parts by weight

Glyceres-26 1.00 parts by weight

arbutin 2.00 parts by weight

Hydrogenated Castor Oil 1.00 parts by weight

ethanol 30.0 parts by weight

preservative a very small amount

coloring agent a very small amount

flavoring agent a very small amount

Purified water remaining amount

<Cosmeceutical Preparation Example 2> Preparation of nourishing cream

Weissella siberia lysate 10.0 parts by weight

1,3-butylene glycol 7.00 parts by weight

glycerin 1.00 parts by weight

D-Panthenol 0.10 parts by weight

plant extract 3.20 parts by weight

Magnesium Aluminum Silicate 0.30 parts by weight

PEG-40 Stearate 1.20 parts by weight

stearic acid 2.00 parts by weight

Polysorbate 60 1.50 parts by weight

Lipophilic Glyceryl Stearate 2.00 parts by weight

Sorbitan sesquioleate 1.50 parts by weight

cetearyl alcohol 3.00 parts by weight

mineral oil 4.00 parts by weight

squalane 3.80 parts by weight

Carlylic/Capric Triglycerides 2.80 parts by weight

vegetable oil 1.80 parts by weight

dimethicone 0.40 parts by weight

dipotassium glycyrrhizate a very small amount

allantoin a very small amount

Sodium Hyaluronate a very small amount

Tocopheryl Acetate appropriate amount

triethanolamine appropriate amount

preservative appropriate amount

flavoring agent appropriate amount

Purified water remaining amount

Manufacturing example of health food

The Weissella civaria lysate according to the present invention can be prepared into various types of health food depending on the purpose. The following exemplifies the manufacturing method of several health foods containing the Weissella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Health food production example 1> Production of dairy products

0.01-1 parts by weight of the Weissella civaria lysate of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<Health food production example 2> Preparation of wire food

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder. Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared into powder having a particle size of 60 mesh with a grinder. The Wasella civaria lysate of the present invention was concentrated under reduced pressure in a vacuum concentrator to obtain a dry powder. It was prepared by mixing the dry powder of the grains, seeds and Weissella civaria lysate prepared above in the following ratio.

Grains (34 parts by weight of brown rice, 19 parts by weight of barley, 20 parts by weight of barley),

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame),

Weissella civaria lysate (2 parts by weight),

Reishi (1.5 parts by weight), and

Rehmannia (1.5 parts by weight).

Manufacturing example of health functional food

The Weissella civaria lysate according to the present invention can be prepared into various types of health functional food depending on the purpose. The following exemplifies the manufacturing method of several health functional foods containing the Wasella civaria lysate according to the present invention as an active ingredient, but the present invention is not limited thereto.

<Health functional food manufacturing example 1> Manufacturing of health functional food

Weissella siberia lysate 100 mg

vitamin mixture appropriate amount

vitamin A acetate 70 μg

vitamin E 1.0 mg

vitamin B1 0.13 mg

vitamin B2 0.15 mg

vitamin B6 0.5 mg

vitamin B12 0.2 μg

vitamin C 10 mg

biotin 10 μg

Nicotinamide 1.7 mg

folic acid 50 μg

Calcium Pantothenate 0.5 mg

mineral mixture appropriate amount

ferrous sulfate 1.75 mg

zinc oxide 0.82 mg

magnesium carbonate 25.3 mg

monobasic potassium phosphate 15 mg

dicalcium phosphate 55 mg

potassium citrate 90 mg

calcium carbonate 100 mg

magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health functional food, but the composition is mixed in a preferred embodiment, but the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method Then, the granules can be prepared and used in the production of a health functional food composition according to a conventional method.

<Health functional food production example 2> Preparation of health functional beverage

Weissella siberia lysate 100 mg

citric acid 100 mg

oligosaccharide 100 mg

Plum Concentrate 2 mg

Taurine 100 mg

Purified water is added to 500 mL

After mixing the above ingredients according to a conventional health drink manufacturing method, after stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized container, sealed and sterilized, and then refrigerated. used in the manufacture of health beverage compositions of Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demand country, and use purpose.

Claims (11)

Weissella cibaria strain, its culture, its lysate, its extract, or a cosmetic composition for skin wrinkle improvement or elasticity enhancement comprising a cell lysate of its extract or culture. According to claim 1,
The skin wrinkles are a cosmetic composition for improving skin wrinkles or enhancing elasticity, characterized in that due to fine dust. Weissella cibaria strain, its culture, its lysate, its extract or a cosmetic composition for preventing or improving skin aging comprising a cell lysate of the culture. 4. The method of claim 3,
The skin aging is a cosmetic composition for preventing or improving skin aging, characterized in that due to fine dust. 4. The method of claim 1 or 3,
The cosmetic 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, pack, soap, cleansing A cosmetic composition, characterized in that at least one formulation selected from formulations consisting of foam, cleansing lotion, cleansing cream, body lotion and body cleanser. Weissella cibaria strain, its culture, its lysate, its extract, or a health functional food composition for skin wrinkle improvement or elasticity enhancement comprising a cell lysate of its extract or culture. Weissella cibaria strain, its culture, its lysate, its extract or a health functional food composition for preventing or improving skin aging, comprising a cell lysate of its extract or culture. 8. The method according to claim 6 or 7,
The health functional food is a health functional food composition, characterized in that the food in the form of tablets, capsules, pills or liquids. Weissella cibaria strain, its culture, its lysate, its extract, or a health food composition for skin wrinkle improvement or elasticity enhancement comprising a cell lysate of its extract or culture. Weissella cibaria ( Weissella cibaria ) A health food composition for preventing or improving skin aging comprising a cell lysate of a strain, its culture, its lysate, its extract or culture. 11. The method of claim 9 or 10,
The health food is a health food composition, characterized in that it is selected from various drinks, meat, sausage, bread, candy, snacks, noodles, ice cream, dairy products, soup, ionic beverages, beverages, alcoholic beverages, gum, tea and vitamin complexes.

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