CN117442510A

CN117442510A - Anti-wrinkle composition based on Sirt signal channel, and preparation method and application thereof

Info

Publication number: CN117442510A
Application number: CN202311439515.7A
Authority: CN
Inventors: 肖永坤; 王东明; 李楚忠; 刘春莹
Original assignee: San Yi Technology Guangzhou Co ltd; Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Current assignee: San Yi Technology Guangzhou Co ltd; Guangzhou Huanya Cosmetic Science and Technology Co Ltd
Priority date: 2023-11-01
Filing date: 2023-11-01
Publication date: 2024-01-26

Abstract

The invention provides a preparation method of an anti-wrinkle composition based on a Sirt signal channel, which comprises the steps of strain culture, enzyme production by fermentation, enzyme extraction, enzyme reaction and purification; the strain is lactococcus lactis subspecies lactis. The fermentation produces enzyme: the fermentation temperature is 36-38 ℃, and after culturing for 24-28 hours, the inducer is added for continuous culturing for 24-36 hours, thus obtaining the enzyme-containing culture solution. The inducer comprises ginsenoside Rb3, lentinus Edodes extract, cordyceps extract, and radix Glehniae extract. The anti-wrinkle composition prepared by the invention has excellent anti-aging effect, can be used for preparing cosmetics, and has obvious anti-wrinkle and anti-wrinkle effects.

Description

Anti-wrinkle composition based on Sirt signal channel, and preparation method and application thereof

Technical Field

The invention relates to the technical field of skin care, in particular to an anti-wrinkle composition based on Sirt signal channels, and a preparation method and application thereof.

Background

With increasing importance of skin care, functional cosmetics face unprecedented opportunities, so that finding cosmetic raw materials with anti-wrinkle, anti-aging and other functions becomes the biggest pain point in the cosmetic industry, and is also the biggest requirement. As a pain in Chinese herbal medicine, ginseng plays an excellent role in wrinkle resistance, and thus, its active ingredient is also a potential candidate with great development value. However, research shows that natural protopanaxoside contained in ginseng belongs to polysaccharide-based ginsenoside, is difficult to directly penetrate cell membranes and be absorbed, and after the ginseng product is orally taken, the polysaccharide-based ginsenoside is degraded into low-sugar-based ginsenoside metabolites in digestive juice and intestinal bacteria in the digestive system, and then can be absorbed and utilized. When the ginseng product is externally used, intestinal bacteria capable of metabolizing ginsenoside are not arranged on the skin, and the ginsenoside exists in a polysaccharide-based prototype, so that the availability is extremely low. Therefore, the modification of the polysaccharide-based saponin into the low-sugar-based saponin has extremely important significance for the development of external products.

Currently, there are mainly four aspects of the mechanism of anti-skin aging: inflammatory signaling pathways, nrf2 signaling pathways, MAPK signaling pathways, and NF- κb signaling pathways, common factors that cause skin aging, such as UVB, ROS, exposure groups, etc., are all associated with these four signaling pathways. However, sirt signaling pathways, which have important effects on cellular aging and are known as "longevity genes", have been less studied for their use in combating skin aging. The Sirt signal channel has important regulation effect on the material metabolism and the energy metabolism of cells, has direct or indirect connection with the aging of the cells through various ways, and is a potential important way for developing anti-skin aging and anti-wrinkle technologies. A schematic of the mechanism by which Sirt signaling channels affect cell senescence is shown in fig. 8. Therefore, the development of products acting on Sirt signal channels is of great importance for the development of techniques against skin aging.

In the prior art, no product for realizing anti-aging and wrinkle removal by regulating the expression of a Sirt signal channel exists.

Disclosure of Invention

Aiming at the technical problems, the invention provides an anti-wrinkle composition based on a Sirt signal channel, a preparation method and application thereof, and aims to achieve the following purposes: promoting Sirt1 expression and realizing remarkable anti-wrinkle and anti-wrinkle effects.

In order to achieve the aim of the invention, the invention adopts the following technical scheme:

a method of preparing an anti-wrinkle composition based on Sirt signaling pathways comprising the steps of:

step 1, strain culture

Inoculating lactococcus lactis subspecies into a liquid culture medium under the conditions of normal temperature and normal pressure; the formula of the liquid culture medium is as follows:

yeast extract 7.5 g, glucose 10.0 g, tomato juice (pressed juice) 100.0 mL, protein wine 7.5 g, KH ₂ PO ₄ 2.0g, tween 800.5 mL, distilled water 900.0 mL, pH 7.0.

The lactococcus lactis subspecies: the strain is commercially available and the strain preservation number is CICC 20402.

After inoculation, the culture is carried out for 35 to 38 hours under the condition of 37 ℃ and oxygen.

Step 2, fermenting to produce enzyme

After 36 hours of cultivation, the culture broth was transferred to a fermentation medium with an inoculum size of 1-2%.

The fermentation medium is as follows: 40-44g of ginseng total saponins, 4.5-6g of silkworm chrysalis meal, 3.5-5g of cow milk meal, 1.8-2.2g of soybean isoflavone, 1.8-2.3g of mannitol, 0.9-1.3g of diammonium citrate, 0.9-1.1g of ascorbic acid, 0.8-1.2g of vitamin B and K ₂ HPO ₄ 0.25-0.35g、ZnSO ₄ ·7H ₂ O 0.15-0.25g、MnSO ₄ ·H ₂ O 0.05-0.06g、（NH ₄ ）SO ₄ ·H ₂ O 0.05-0.06g、Ca（NO ₃ ） ₂ 0.8-1.2g, fucoidin 5-7g, glutamic acid 0.8-1.2g, distilled water 1.0L, pH6.8-7.0. Culturing at 36-38deg.C for 24-28 hr, adding inducer, shaking the culture medium, and culturing for 24-36 hr to obtain enzyme-containing culture solution.

The inducer comprises ginsenoside Rb3, lentinus Edodes extract, cordyceps extract, and radix Glehniae extract according to the ratio of 1-3:1-2:2-3:1 to 1.4 mass ratio.

Step 3, extracting enzyme

After fermentation, the thalli are removed by centrifugation, and then the fermentation liquor is concentrated by an ultrafiltration membrane to remove impurities such as protein, salt, sugar and the like with molecular weight lower than 20kDa, thus obtaining crude enzyme.

Step 4, enzyme reaction

Dissolving crude enzyme in phosphate buffer solution with pH of 7.0 and 0.05M, wherein the mass ratio of the crude enzyme to the phosphate buffer solution is 1:48-52 to obtain the enzyme solution.

Rb3 with the mass percentage of 40% is added into a phosphate buffer solution with the concentration of 0.05M and the pH value of 7.0, and the mixture is fully dissolved to obtain a substrate. The mass ratio of Rb3 to phosphate buffer is 1:15-25. The Rb3: is commercially available.

Mixing the enzyme solution and the substrate solution according to the mass ratio of 1:1, and reacting for 18-26h at 35-38 ℃.

Step 5, purification

After fermentation, filtering to remove thalli, concentrating the culture solution to 5-20% of the volume of the culture solution to obtain concentrated solution. Adding 3-4 times of ethanol into the concentrated solution, oscillating for 20-30min, and centrifuging to retain centrifugate. The ethanol concentration was 95%.

Purifying the centrifugate with macroporous adsorption resin; after the saponin is adsorbed by the resin, washing to remove impurities; washing the column with 50% ethanol to remove polysaccharide-based saponin; then eluting the saponin with 95% alcohol, evaporating the eluent to dryness to obtain powder, namely the anti-aging composition.

An anti-wrinkle composition based on Sirt signaling pathways comprising the following components: ginsenoside CMc, ginsenoside CMx, and ginsenoside CK; the mass ratio is CMc: CMx: ck=1:10-15:3-4.

Compared with the traditional method, the invention has the following beneficial effects:

the anti-wrinkle composition based on the Sirt signal channel has an excellent anti-aging effect, can be used for preparing cosmetics such as emulsion, essence, nutrient solution, face cream and the like, can obviously promote the expression of Sirt1 protein, and has obvious anti-wrinkle and anti-wrinkle effects.

The nutrient solution prepared by the anti-wrinkle composition is used for skin care, and the number, the area and the length of wrinkles can be obviously reduced.

Drawings

FIG. 1 is an HPLC detection chart of the product of example 1;

FIG. 2 is an electrophoresis chart of the detection of liver Sirt1, FOXO1, PPARα, PGC1- α proteins in example 3;

FIG. 3 is a graph showing the analysis of Sirt1 content in the liver of the rat in example 3;

FIG. 4 is a graph showing the analysis of the amount of factor FOXO1 in the liver of the rat in example 3;

FIG. 5 is a graph showing the analysis of specific levels of peroxisome proliferator-activated receptor PPARα in the liver of rats in example 3;

FIG. 6 is a graph showing the analysis of specific content of hydrogen effector PGC 1-alpha in the liver of rats in example 3;

FIG. 7 is a photograph of Primos-CR canthus wrinkles in the test area of volunteers at days 0, 28, 56 of example 4;

FIG. 8 is a schematic of the mechanism by which Sirt signaling channels affect cell senescence.

Detailed Description

Example 1A method for preparing an anti-wrinkle composition based on Sirt signaling channels

The method comprises the following steps:

step 1, strain culture

After inoculation, the cells were incubated at 37℃for 36 hours under aerobic conditions.

Step 2, fermenting to produce enzyme

The fermentation medium is as follows: 42.0g of ginseng total saponin, 5.0g of silkworm chrysalis meal, 4.0g of cow milk powder, 2.0g of soybean isoflavone, 2.0g of mannitol, 1.0g of citric acid diammonium, 1.0g of ascorbic acid, 12.0 g of vitamin B and K ₂ HPO ₄ 0.3g、ZnSO ₄ ·7H ₂ O 0.2g、MnSO ₄ ·H ₂ O 0.05g、（NH ₄ ）SO ₄ ·H ₂ O 0.05g、Ca（NO ₃ ） ₂ 1.0g of fucoidan, 6.0g of glutamic acid, 1.0L of distilled water and pH6.9. At 37 ℃ for 26 hours, then adding an inducer, shaking the culture medium uniformly after adding, and continuing to culture for 28 hours with the inducer added amount being 3% of the mass of the expanded culture medium to obtain the enzyme-containing culture solution.

The inducer comprises ginsenoside Rb3, lentinus Edodes extract, cordyceps extract, and radix Glehniae extract according to the following ratio of 2:1.5:3: 1.2, mixing.

Step 3, extracting enzyme

Step 4, enzyme reaction

Dissolving crude enzyme in phosphate buffer solution with pH of 7.0 and 0.05M, wherein the mass ratio of the crude enzyme to the phosphate buffer solution is 1:50, obtaining an enzyme solution.

Adding 40 mass percent of Rb3 into 0.05M phosphate buffer with pH of 7.0, and fully dissolving to obtain the substrate.

The mass ratio of Rb3 to phosphate buffer is 1:20. The Rb3: is commercially available.

The enzyme solution and the substrate are mixed according to the mass ratio of 1:1 and reacted for 20 hours at 37 ℃.

Step 5, purification

After fermentation, the bacterial cells are removed by filtration, and then the culture solution is concentrated to 10% of the volume of the culture solution, so as to obtain a concentrated solution. Adding 4 times of ethanol into the concentrated solution, oscillating for 20-30min, and centrifuging to retain centrifugate. The ethanol concentration was 95%.

Purifying the centrifugate with macroporous adsorption resin; after the saponin is adsorbed by the resin, washing to remove impurities; washing the column with 50% ethanol to remove polysaccharide-based saponin; then eluting the saponin with 95% alcohol, evaporating the eluent to dryness to obtain the product, namely the anti-aging composition. The product was subjected to HPLC detection, the HPLC detection pattern is shown in FIG. 1.

The detected product components comprise ginsenoside CMc, CMx, CK in mass ratio: CMc: CMx: ck=1:12:3.6.

Example 2 use of an anti-wrinkle composition based on Sirt signaling channels

Dissolving the anti-aging composition prepared in the example 1 in a solvent to prepare an anti-wrinkle nutrient solution; the mass ratio of the anti-aging composition to the solvent is 1:200.

The solvent is prepared from water, ethanol, glycerol and squalane according to the following ratio of 80:3:12:5 mass ratio.

Example 3 test of Effect of promoting protein expression on Sirt1 Signal pathway

The detection method comprises the following steps:

step 1, animal modeling grouping and administration

After the Wistar rats were adaptively fed for 1 week, they were randomly divided into 3 groups, 1 group was selected as a normal group, and the remaining 2 groups were subjected to modeling. The normal group was fed with normal feed for 6 weeks. The building block was fed with high fat diet for 6 weeks. 6. The rats were modeled after week on an empty stomach and were intraperitoneally injected with STZ 6mg/kg (dissolved in sodium citrate buffer, pH 4.5) to create a T2DM model. The normal group was intraperitoneally injected with the same amount of sodium citrate buffer as the modeling group. After injection 3 d, the random blood sugar is measured to be more than or equal to 16.7 mol/L, which indicates that the molding is successful. Rats with substandard blood sugar are continuously observed according to the previous steps after being subjected to intraperitoneal injection of 5 mg/kg STZ again, and if the blood sugar is not qualified after intraperitoneal injection for 2 times, the rats are killed under the anesthesia of the rejection molding module. After molding the model rats, they were randomly divided into model groups and drug administration groups.

Drug administration group: the composition prepared in example 1 was dissolved in distilled water to prepare a 0.1/mL solution, which was administered by gavage at a dose of 0.4. 0.4 g/kg. Rats in the normal and model groups were perfused with equal volumes of distilled water.

1. The stomach was irrigated continuously for 6 weeks at times/d.

Step 2, detecting liver Sirt1, FOXO1, PPARα, PGC1- α protein expression

Livers of rats in the normal group, model group, and administration group were collected. Cutting liver tissue, cleaning in 2 mL EP tube, adding 300L RIPA lysate, repeatedly grinding, and performing ice lysis for 10 min at 4deg.C. The supernatant was collected by centrifugation at 12000 r/min radius 8 cm for 15 min and protein concentration was determined by BCA protein quantification. Preparing 10% of separation gel and 4.8% of concentrated gel, adding TEMED, shaking and filling the gel, adding a total protein sample with a corresponding volume, mixing with 5x protein gel electrophoresis loading buffer solution, boiling for denaturation, transferring to PVDF membrane, sealing in sealing solution for 90 min, adding Sirt1, FOXO1, PPARα and PGC1- α antibodies, standing overnight at 4 ℃, washing with TBST for 3 times, 10 min each time, adding secondary antibody for incubation for 90 min, washing the membrane, incubating ECL chemiluminescent solution with the membrane for 1min, imaging by a gel imaging system, and detecting the result is shown in figure 2.

The analysis chart of Sirt1 content in livers of rats in the normal group, the model group and the administration group is shown in figure 3; the analysis chart of the content of the transcription factor FOXO1 in the livers of the normal group, the model group and the administration group rats is shown in figure 4; the specific content analysis chart of peroxisome proliferator-activated receptor PPARα in the livers of the normal group, model group and administration group rats is shown in FIG. 5; the analysis of the specific content of the hydrogen effector molecule PGC 1-alpha in the livers of the rats in the normal group, the model group and the administration group is shown in FIG. 6.

Analysis of FIG. 2 shows that the levels of Sirt1, FOXO1, PPARα, and PGC1- α were significantly lower in the model group than in the normal group, indicating that expression of Sirt1, FOXO1, PPARα, and PGC1- α was affected in the rats after modeling. The levels of Sirt1, FOXO1, pparα, PGC1- α were significantly higher in the administered group than in the model group, demonstrating that intervention of the saponin composition prepared in example 1 effectively increased the expression of Sirt1, FOXO1, pparα, PGC1- α in rats, i.e., the saponin composition prepared in example 1 significantly upregulated the expression of proteins on the Sirt1 signaling pathway.

As can be seen from FIG. 2, the amount of protein expressed on the Sirt1 signal pathway in the rats of the administration group was 2 times or more that in the model group.

Example 4 anti-wrinkle nutrient solution use Effect experiment

30 volunteers were recruited for the anti-wrinkle nutrient solution trial experiment prepared in example 2, numbered VT001-VT056, respectively; from the first day of the experiment, volunteers used anti-wrinkle nutrient solution for skin care three times a day in the morning, in the middle and at night, without using other skin care products at the same time during the experiment. The number of Primos-CR canthus wrinkles, the area of wrinkles, and the length of wrinkles in the tested area of volunteers were measured and counted daily for 56 days. Photographs of Primos-CR canthus wrinkles of the tested area of VT001, VT006, VT026 volunteers at days 0, 28, 56 were taken at random, as shown in FIG. 7.

1. The average value of the number of wrinkles at the corners of the eyes of the volunteer was measured and calculated before the experiment and was taken as a base value D0. On day 28 of the product, the average D28 of the number of wrinkles at the corners of the volunteer's eyes was detected and calculated. On day 56 of the product, the average D56 of the number of wrinkles at the corners of the volunteer's eyes was detected and calculated. The results (mean.+ -. Standard error) of the number of Primos-CR canthus wrinkles in the volunteer test areas at day 0, 28, 56 of the experiment are shown in Table 1;

TABLE 1

Note that: data in the table are mean ± standard error

Calculating the change rate of the number of wrinkles at the canthus for 28 days using the product and the change rate of the number of wrinkles at the canthus for 56 days using the product, as shown in table 2;

rate of change in number of canthus wrinkles on day n= (average number of canthus wrinkles after day N using product-average number of canthus wrinkles on day 0) ×100%/average number of canthus wrinkles on day 0

TABLE 2

The significance of the results of the number of wrinkles at the corners of the eyes of the volunteers in this experiment is shown in Table 3;

TABLE 3 Table 3

Note that: statistical method analysis was performed using a rank sum test and t test method, with test level a=0.05.

The significance labeling method is that "n.s" indicates no statistical difference, p >0.05, p <0.05 indicates significant difference (". Times." indicates that 0.01.ltoreq.p <0.05; ". Times." indicates that 0.001.ltoreq.p <0.01; ". Times." indicates that p < 0.001), and the number of products is used=30.

As shown in tables 1 to 3, after the volunteers continuously use the product anti-wrinkle nutrient solution for 28 days and 56 days, the number of Primos-CR canthus wrinkles in the tested area is remarkably reduced (p < 0.001) compared with the basic value, and the reduction rates are 27.52% and 44.97% respectively. The anti-wrinkle nutrient solution provided by the invention can effectively reduce the number of wrinkles, and has a remarkable anti-aging effect.

2. The average value of the volunteer's canthus wrinkle volume was measured and calculated prior to the experiment and taken as the base value D0. After 28 days of use of the product, the average D28 of the volunteer's corner of eye wrinkle volume was detected and calculated. After 56 days of use of the product, the average D56 of the volunteer's corner of eye wrinkle volume was detected and calculated. The results of the test (mean.+ -. Standard error) of the volume of Primos-CR canthus wrinkles in the volunteer test area at day 0, 28, 56 of the experiment are shown in Table 4;

TABLE 4 Table 4

Note that: data in the table are mean ± standard error

Calculating the change rate of the volume of the canthus wrinkles in 28 days of using the product and the change rate of the volume of the canthus wrinkles in 56 days of using the product, as shown in table 5;

rate of change in volume of corner of eye wrinkles on day n= (average of corner of eye wrinkles after day N using product-average of corner of eye wrinkles on day 0) ×100%/average of corner of eye wrinkles on day 0

TABLE 5

The significance of the results of the volume detection of the wrinkles at the corners of the eyes of the volunteers in this experiment is shown in Table 6;

TABLE 6

As can be seen from tables 4 to 6, volunteers continuously used the product anti-wrinkle nutrient solution for 28 days, the Primos-CR canthus wrinkle volume in the tested area was significantly reduced (0.01 Sp < 0.05) from the basal value, the reduction rate was 11.67%, and the Primos-CR canthus wrinkle volume in the tested area was significantly reduced (p < 0.001) from the basal value, the reduction rate was 17.56% after the volunteers continuously used the product anti-wrinkle nutrient solution for 56 days.

3. The average value of the length of the wrinkles at the corners of the eyes of the volunteers was measured and calculated before the experiment and was taken as a base value D0. After 28 days of use of the product, the average value D28 of the length of the wrinkles at the corners of the eyes of the volunteers was detected and calculated. After 56 days of use of the product, the average value D56 of the length of the wrinkles at the corners of the eyes of the volunteers was detected and calculated. The results of the test (mean.+ -. Standard error) of the length of Primos-CR canthus wrinkles in the volunteer test areas at day 0, 28, 56 of the experiment are shown in Table 7;

TABLE 7

Note that: data in the table are mean soil standard error

Calculating the change rate of the length of the canthus wrinkle in 28 days of using the product and the change rate of the length of the canthus wrinkle in 56 days of using the product, as shown in table 8;

rate of change in length of corner wrinkles at day n= (average value of corner wrinkles after day N using product-average value of corner wrinkles at day 0) ×100%/average value of corner wrinkles at day 0

TABLE 8

The significance of the results of the test volunteer's canthus wrinkle length is shown in table 9;

TABLE 9

As shown in tables 7-9, the Primos-CR eye corner wrinkle length of the tested area in 28 days and 56 days of continuous use of the product anti-wrinkle nutrient solution by the volunteers is remarkably reduced (p is more than or equal to 0.001 and less than or equal to 0.01) compared with the base value, and the reduction rates are respectively 10.21 percent and 11.34 percent.

The control conditions for HPLC detection described in the present invention are as follows: the detection wavelength is 203nm. Mobile phase acetonitrile (a) -water (B): 0-20min,20% A,20-31min, 20-32% A,31-40min, 32-43% A,40-47min, 43-100% A. Column temperature was 35 ℃.

The raw materials adopted by the invention are all from commercial sources. The ginseng total saponins: the saponin content is 80%, the product model is HS-376, the gramineous organism; the Lentinus edodes extract comprises the following components: polysaccharide content 50%, model WYT; the Cordyceps extract comprises: polysaccharide content 30%, product model RC2303008; the radix glehniae extract: specification 30:1, product model BSSTQW01.

Except for specific descriptions, the proportions of the invention are mass ratios, and the percentages are mass percentages.

It is apparent that there are many more specific embodiments that can be varied within the inventive concept, and it should be stated herein that any modification made within the inventive concept of the present invention will fall within the scope of the present invention.

Claims

1. A method of preparing an anti-wrinkle composition based on Sirt signaling pathways, comprising: comprises the steps of strain culture, enzyme production by fermentation, enzyme extraction, enzyme reaction and purification; the strain is lactococcus lactis subspecies lactis.

2. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 1, wherein: the fermentation produces enzyme: the fermentation temperature is 36-38 ℃, and after culturing for 24-28 hours, the inducer is added for continuous culturing for 24-36 hours, thus obtaining the enzyme-containing culture solution.

3. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 2, characterized in that: the inducer comprises ginsenoside Rb3, lentinus Edodes extract, cordyceps extract, and radix Glehniae extract.

4. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 3, wherein: the proportion of the ginsenoside Rb3, the mushroom extract, the cordyceps extract and the glehnia root extract is 1-3:1-2:2-3:1-1.4.

5. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 1, wherein: the enzyme reaction: mixing the enzyme solution with the substrate, and reacting at 35-38 ℃ for 18-26h.

6. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 1, wherein: the enzyme reaction: dissolving crude enzyme in phosphate buffer solution, wherein the mass ratio of the crude enzyme to the phosphate buffer solution is 1:48-52 to obtain the enzyme solution.

7. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 1, wherein: the enzyme reaction: adding Rb3 into phosphate buffer solution, and fully dissolving to obtain a substrate; the mass ratio of Rb3 to phosphate buffer is 1:15-25.

8. A method of preparing an anti-wrinkle composition based on Sirt signal pathways according to claim 1, wherein: the fermentation produces enzyme: the fermentation medium adopted comprises the following components: total ginsenoside, pupa Bombycis powder, milk powder, soybean isoflavone, mannitol, diammonium citrate, ascorbic acid, vitamin B12, K ₂ HPO ₄ 、ZnSO ₄ ·7H ₂ O、MnSO ₄ ·H ₂ O、（NH ₄ ）SO ₄ ·H ₂ O、Ca（NO ₃ ） ₂ Fucoidan, glutamic acid.

9. An anti-wrinkle composition based on Sirt signaling pathways, characterized by: the composition comprises the following components: ginsenoside CMc, ginsenoside CMx, and ginsenoside CK.

10. An anti-wrinkle composition based on Sirt signaling pathway according to claim 9, wherein: the mass ratio of the ginsenoside CMC to the ginsenoside CMx to the ginsenoside CK is 1:10-15:3-4.