KR102218224B1 - Cosmetic composition for preventing or improving skin inflammation or skin aging comprising fermented product of Ligularia stenocephala leaf as an active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for the prevention or improvement of skin inflammation or skin aging containing a fermented product of gondalbee leaf as an active ingredient, specifically Bacillus subtilis in a liquid medium containing the powder of gondalbee leaf , Lactobacillus plantarum (Lactobacillus plantarum) and Saccharomyces cerevisiae (Saccharomyces cerevisiae) inoculation and fermentation of the obtained fermented product containing as an active ingredient for the prevention or improvement of skin inflammation or skin aging cosmetic composition About.
The cosmetic composition of the present invention may exhibit excellent anti-inflammatory activity and antioxidant activity by containing a fermented product of gondalbee leaf obtained through a specific fermentation as an active ingredient, thereby exhibiting excellent effects in preventing or improving skin inflammation or skin aging.

Description

Cosmetic composition for preventing or improving skin inflammation or skin aging comprising fermented product of Ligularia stenocephala leaf as an active ingredient}

The present invention relates to a cosmetic composition for the prevention or improvement of skin inflammation or skin aging containing a fermented product of gondalbee leaf as an active ingredient, specifically Bacillus subtilis in a liquid medium containing the powder of gondalbee leaf , Lactobacillus plantarum (Lactobacillus plantarum) and Saccharomyces cerevisiae (Saccharomyces cerevisiae) inoculation and fermentation of the obtained fermented product containing as an active ingredient for the prevention or improvement of skin inflammation or skin aging cosmetic composition About.

China, Beijing, India and Korea are currently in a very serious stage of fine dust, and these fine dusts lead to skin diseases as well as respiratory diseases for all modern people, causing various types of skin diseases such as contact dermatitis and eczema, and health and life. Is showing serious damage to

In the case of the steadily growing overseas and domestic cosmetics industry and, in particular, in the'Derma Cosmetics' market, which combines skin science with cosmetics, the market size of soothing products for sensitive skin-related troubles caused by fine dust, etc. is 500 billion won, growing more than 15% annually. have.

Fermentation refers to the process of decomposing organic matter using microbial enzymes, and the fermentation method, the oldest technology in history, is widely used in various fields such as food, medicine, and cosmetics. Among them, food fermentation is not only in Korea but also worldwide. It is the most widely used. In the case of food fermentation, it is known that it has the effect of improving the flavor, flavor, and texture of food through the enzyme action of microorganisms as a traditional method of processing refinement, destroying toxic substances through fermentation, and enhancing physiologically active substances. .

On the other hand, gondalbi, which is grown or cultivated in Korea, is a wild vegetable similar to Gomchwi, and is mainly used as a food ingredient, and has not yet been used in beauty products such as cosmetics.

The inventors of the present invention attempted to utilize this gondalbi in cosmetics, developed a fermentation method to obtain a fermented product with excellent physiological activity by using gondalbi as a raw material, and developed a cosmetic composition having excellent functionality by applying the fermented product to cosmetics I wanted to.

Korean Patent Registration No. 10-1694229

The main object of the present invention is to provide a cosmetic composition exhibiting an excellent effect in preventing or improving skin inflammation or skin aging by containing a fermented product obtained through specific fermentation of Gondalbi.

According to one aspect of the present invention, the present invention is Bacillus subtilis (Bacillus subtilis), Lactobacillus plantarum (Lactobacillus plantarum) and Saccharomyces cerevisiae (Saccharomyces cerevisiae) in a liquid medium containing the powder of Gondal Bee leaf. ) It provides a cosmetic composition for preventing or improving skin inflammation or skin aging containing a fermented product obtained by inoculation and fermentation as an active ingredient.

In the cosmetic composition of the present invention, the liquid medium contains 10 to 30 g/L of the gondalbee leaf powder, and it is preferable that sugar is further contained.

In the cosmetic composition of the present invention, the ferment is inoculated with Bacillus subtilis , Lactobacillus plantarum and Saccharomyces cerevisiae in the liquid medium, and 30 It is preferably obtained by shaking fermentation for 2 to 4 days at 100 to 200 rpm at 40°C.

The cosmetic composition of the present invention may exhibit excellent anti-inflammatory activity and antioxidant activity by containing a fermented product of gondalbee leaf obtained through a specific fermentation as an active ingredient, thereby exhibiting excellent effects in preventing or improving skin inflammation or skin aging.

Figure 1 shows the results of the cytotoxicity test of fermented gondalbi leaf according to an embodiment of the present invention. control, control (no treatment); 10 ~ 2.5mg / ml, treatment group by concentration of the fermented product of the present invention; Gondalbi fermentation 0d, before fermentation; Gondal fermentation 3d, 3 days after fermentation; Gondal fermentation 7d, 7 days after fermentation.
Figure 2 shows the experimental results of the NO production inhibitory effect of fermented gondalbi leaf according to an embodiment of the present invention. control, control (no treatment); LPS10ng, LPS (10ng/ml) treatment group; Gondalbi 0d1~0.25mg, treatment group by concentration of the mixture before fermentation of Gondalbi leaf; Gondalbi 3d1~0.25mg, treatment group by concentration of fermented fermented gondalbee leaves for 3 days; Gondalbi 7d1~0.25mg, treatment group of fermented products fermented with gondalbi leaves for 7 days by concentration.
Figure 3 shows the experimental results of the NO production inhibitory effect of fermented gondalbi leaf according to an embodiment of the present invention. control, control (no treatment); LPS10ng/ml, LPS(10ng/ml) treatment group; Gondalbi 0d1~0.25mg/ml, treatment group by concentration of the mixture before fermentation of Gondalbi leaf; Gondalbi 3d1~0.25mg/ml, treatment group by concentration of fermented fermented gondalbi leaves for 3 days; Gondalbi 7d1~0.25mg/ml, treatment group by concentration of fermented fermented gondalbee leaves for 7 days.
Figure 4 shows the total polyphenol content of fermented gondalbi leaf according to an embodiment of the present invention. Gondalbi 0d10~2.5mg/ml, treatment group by concentration of the mixture before fermentation of Gondalbi leaf; Gondalbi 3d10~2.5mg/ml, treatment group by concentration of fermented fermented gondalbee leaves for 3 days; Gondalbi 7d10~2.5mg/ml, treatment group by concentration of fermented fermented gondalbee leaves for 7 days.
Figure 5 shows the total flavonoid content of fermented gondalbi leaf according to an embodiment of the present invention. Gondalbi 0d5~1.25mg/ml, treatment group by concentration of the mixture before fermentation of Gondalbi leaf; Gondalbi 3d5~1.25mg/ml, treatment group by concentration of fermented fermented gondalbi leaves for 3 days; Gondalbi 7d5~1.25mg/ml, treatment group by concentration of fermented fermented gondalbee leaves for 7 days.
Figure 6 shows the DPPH radical scavenging activity of fermented gondalbi leaf according to an embodiment of the present invention. 10 ~ 2.5mg / ml, treatment group by concentration of the fermentation of the present invention; VC100~10ug/ml, treatment group by vitamin C concentration; Gondalbi fermentation 0d, before fermentation; Gondal fermentation 3d, 3 days after fermentation; Gondal fermentation 7d, 7 days after fermentation.

The cosmetic composition of the present invention is fermented by inoculating Bacillus subtilis , Lactobacillus plantarum , and Saccharomyces cerevisiae in a liquid medium containing the powder of Gondalbee leaf. It is characterized by containing the obtained fermented product as an active ingredient.

In the present invention, the powder of the gondalbi leaf can be obtained through a conventional method of pulverizing the plant material, but it is preferable to use a powder obtained by lyophilizing the green leaf of the gondalbi in order to increase the effect of the present invention.

In the present invention, the liquid medium can be obtained by adding nutrients necessary for the fermentation bacteria to survive during the fermentation period together with the powder of the gondalbi leaf to water. The liquid medium contains 10 to 30 g/L of the gondalbee leaf powder, and it is preferable that sugar is further contained as an additional nutrient. At this time, it is preferable to use glucose and sucrose as the sugar, and it is preferable to contain 1 to 10 g/L of glucose and 1 to 10 g/L of sucrose in the liquid medium.

The liquid medium is preferably sterilized before inoculation of the fermenting bacteria in order to prevent erroneous fermentation due to the growth of various bacteria that may occur during the fermentation process. At this time, the sterilization may use a conventional liquid sterilization method, but it is preferable to use a method of high temperature and high pressure heating.

In the present invention, Bacillus subtilis, Lactobacillus plantarum and Saccharomyces cerevisiae are used as fermenting bacteria, in which case KCCM 11316, ATCC 6633, IFO 3134, DSM 347 or NRRL B-209 are used as Bacillus subtilis strains. It is preferable to use, and it is preferable to use KCCM 11322, ATCC 8014, IFO 3070, DSM 20205 or NRRL B-531 as the Lactobacillus plantarum strain, and KCCM 11215, IFO as the Saccharomyces cerevisiae strain. It is preferred to use 1833 or NRRL Y-12637.

In the present invention, the inoculation may be performed by adding the pre-culture solution of the fermenting bacteria to the liquid medium. At this time, the pre-culture solution of the fermenting bacteria can be obtained by inoculating and culturing the fermenting bacteria in a liquid medium in which the fermenting bacteria can grow. At this time, Bacillus subtilis is preferably cultured in TSB liquid medium at 30 to 40°C and 100 to 200 rpm for 1 to 3 days, and Lactobacillus plantarum is cultured in MRS liquid medium. It is preferable to use a method of shaking culture at 40℃, 100 ~ 200rpm for 1-3 days, and culture of Saccharomyces cerevisiae is performed in YEPD liquid medium by shaking for 1-3 days at 30 ~ 40℃ and 100 ~ 200rpm It is preferable to use the method of culturing. In order to increase the fermentation efficiency, it is preferable to inoculate the pre-culture solution obtained in this way by adding 0.5 to 2% (v/v) per strain culture solution.

In the present invention, the fermentation may be performed by maintaining the liquid medium containing the gondalbee leaf powder inoculated with the fermenting bacteria at a temperature capable of growth and fermentation of the fermenting bacteria for an appropriate time. At this time, the temperature is preferably 30 ~ 40 ℃, it is desirable to ferment shaking for 2 ~ 4 days at 100 ~ 200rpm.

In the present invention, the fermented product prepared through the above method can be used as it is, but it is preferable to use it after removing the solid material and lyophilizing it. At this time, the solids may be removed by conventional methods such as sedimentation, filtration, and centrifugation.

The cosmetic composition of the present invention is a form commonly prepared in the art, for example, skin, lotion, cream, essence, nutrient water, lotion, pack, soap, shampoo, rinse, cleansing, body cleanser, face wash, treatment and essence It may be prepared in the form of, and may be prepared by adding the fermented product during the manufacturing process of these types of cosmetics. At this time, the amount of fermented product added may be appropriately selected in consideration of the type, shape, purpose, and manufacturing method of the cosmetic, and for example, the fermented product may contain 0.1 to 20% by weight based on the weight of the total cosmetic composition. have.

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

[Example]

1. Manufacture of fermented gondalbee leaf

1-1. Preparation of fermentation microorganisms

Bacillus subtilis (KCCM 11316), Lactobacillus plantarum (KCCM 11322) and Saccharomyces cerevisiae (KCCM 11215) were TSA, MRS, YEPD solids, respectively. Plates were plated on the medium and cultured at 36° C. for 2 days, and then each cultured strain 1 platinum was inoculated into 100 ml of TSB, PDB, and YEPD liquid medium, followed by shaking culture at 36° C. for 2 days at 160 rpm.

1-2. Fermentation production

Liquid medium per liter Last name content glucose 5g sucrose 5g Raw Gondalbee Leaf Freeze-Dried Powder 20g Distilled water up to 1000ml

As shown in Table 1, a liquid medium was prepared, sterilized at 121°C for 30 minutes at 1.5psi, cooled, and 10 ml of each strain culture solution of 1-1 was added and inoculated, followed by shaking fermentation at 36°C at 160 rpm for 0 to 7 days. .

Thereafter, the fermentation broth was settled at 4° C. for 24 hours at low temperature, and then centrifuged at 4° C. and 4,500 rpm to obtain a supernatant, and filtered twice with a filter having a pore size of 5 μm and 0.45 μm, respectively, and the filtrate was freeze-dried.

2. Cytotoxicity test

The cytotoxicity of the fermented product (lyophilized powder) sample of Example 1 was investigated.

2-1. Cell culture

RAW264.7 (TIB-71; ATCC, USA) macrophages used in the experiment were purchased from American type cell collection (ATCC, USA), and the medium used was 10% FBS in DMEM (Dulbecco's Modified Eagle's Medium; Gibco, USA) medium. (Fetal Bovine Serum; Gibco, USA) and 1% of Penicillin (100U/ml)/Streptocycin (100µg/ml)/Amphotericin B (0.25µg/ml) were added, and then under the conditions of _{37°C and 5% CO 2} It was used after culturing.

2-2. Cytotoxicity assessment (MTT assay) Cell Viability Assay

The cell viability of the sample was measured by the EZ-Cytox (Enhanced Cell Viability Assay kit, DoGenBio Co., Ltd.) method. The cells were dispensed into a 96-well plate at a concentration of 5×10 ⁴ cells/well and cultured for 24 hours, and then the medium was removed. Add 20 µl of each sample diluted by concentration to 180 µl of new DMEM (10% FBS, 1% Penicillin (100U/ml)/Streptocycin (100µg/ml)/Amphotricin B (0.25µg/ml)). After incubation for time, 10 µl of EZ-Cytox reagent was added to each well and incubated for 1 hour at 37° C. and 5% CO ₂ . After reacting for 1 hour, the absorbance was measured at 450 nm using an ELISA reader.

2-3. result

Experimental results As shown in Fig. 1, the fermented product of Example 1 was found to have no significant cytotoxicity compared to the control at all concentrations.

3. NO production inhibitory effect

The effect of inhibiting NO generation of the fermented product (lyophilized powder) sample of Example 1 was investigated. The NO production inhibitory effect is used as an indicator of the anti-inflammatory effect.

RAW 264.7 cells ^{were dispensed into a 12-well plate at a concentration of 2.5×10 5} cells/ml, cultured for 24 hours, and then the medium was removed. Here, a sample diluted to 1 mg/ml in fresh DMEM (10% FBS, 1% Penicillin (100U/ml)/Streptocycin (100µg/ml) Amphotericin B (0.25µg/ml)) medium was pretreated for 1 hour. Then, LPS (10 ng/ml) was treated and incubated for 24 hours at 37°C and 5% CO ₂ . After 24 hours of sample treatment, the cell culture solution was centrifuged, and 100 µl of the supernatant and 50 µl of Griess reagent A (1% sulfanilamide) + 50 µl of B solution (0.1% N-(1-naphtyl) ethylenediamine dihydrochloride) were mixed to 96 well plates. After reacting at for 20 minutes, the absorbance was measured at 540 nm using an ELISA reader. As a result _{, a standard curve was drawn with 1M NaNO 2,} and the value of NO contained in the cell culture supernatant was calculated.

As a result of this, as shown in FIGS. 2 and 3, both of the fermented products of Example 1 were found to have an excellent inhibitory effect on NO generation, and particularly, when fermented for 3 days, the effect of inhibiting NO generation was the most excellent.

4. Total polyphenol content

The total polyphenol content of the fermented product (lyophilized powder) sample of Example 1 was investigated.

The total polyphenol content was measured using the method of Na Hyun Lee et al. (2015). _{Add 2 ml of 2% Na 2} CO ₃ solution to 100 μl of sample solution (diluted with water to a concentration of 10 ~ 2.5 mg/ml) or Gallic acid standard (0, 20, 40, 60, 80, 100 mg/L) After stirring and allowing to stand for 3 minutes, 50% Folin &Ciocalteu's phenol reagents 100 µl were added and vortexed, reacted in a dark room for 30 minutes, and the absorbance was measured using a blank at 750 nm as a control. To quantify the phenolic compound, it was calculated from a calibration curve obtained using gallic acid as a standard material.

As a result of this, as shown in FIG. 4, it was found that all of the fermented products of Example 1 had a high total polyphenol content.

5. Total flavonoid content

The total flavonoid content of the fermented product (lyophilized powder) sample of Example 1 was investigated.

The total flavonoid content was measured by modifying the method of Hongmei Geng. (2008). After mixing 2 ml of sample solution (diluted with water to a concentration of 5 to 1.25 mg/ml) or Rutin hydrate standard (0, 10, 20, 30, 40, 50 mg/L) and 0.3 _{ml of 5% NaNO 2,} After reacting for 6 minutes, 0.3 ml of 10% Al(NO ₃ ) ₃ was added and reacted for 6 minutes at room temperature. After 4 ml of 10% NaOH solution was added and stirred sufficiently, 10 ml of 2 distilled water was added to the mixture, followed by reaction for 15 minutes. Then, using a spectrophotometer (OPTIZEN POP Meccasys), the blank was compared at 510 nm and the absorbance of each solution was measured. The total flavonoid content was calculated from a calibration curve prepared using Rutin hydrate as a standard material.

As a result of this, as shown in FIG. 5, it was found that all of the fermented products of Example 1 had a high total flavonoid content.

6. Antioxidant effect

The antioxidant effect of the fermented product (lyophilized powder) sample of Example 1 by concentration (10 mg/ml, 5 mg/ml, 2.5 mg/ml, 1.25 mg/ml) was investigated.

The antioxidant effect was investigated by the DPPH radical scavenging activity test method, and the DPPH radical scavenging activity test method modified from the method of Seon-hee you (2016) was used. 0.1mM DPPH (2,2-diphenyl-1-picrylhydrazyl) solution was added to a 96-well plate in an equal amount of 180 µl each, and then 20 µl of samples of various concentrations dissolved in distilled water were added each to react at 37°C for 30 minutes. Absorbance was measured at 517 nm. At this time, ascorbic acid (vitamin C) was used as a control. DPPH radical scavenging activity was calculated by the following equation.

DPPH radical scavenging activity (%) = 100-{(absorbance in added group / absorbance in no added group) × 100}

As a result, as shown in FIG. 6, both fermented products of Example 1 exhibited excellent antioxidant activity.

Claims (3)

Bacillus subtilis (Bacillus subtilis), Lactobacillus plantarum (Lactobacillus plantarum) and saccharin in a liquid medium containing 10 to 30 g/L of fresh gondal leaf freeze-dried powder, 1 to 10 g/L of glucose and 1 to 10 g/L of sucrose Cosmetics for preventing or improving skin inflammation or skin aging containing fermented products obtained by inoculating Saccharomyces cerevisiae and shaking fermentation at 30 to 40°C at 100 to 200 rpm for 2 to 4 days as an active ingredient Composition.
delete delete

Images