CN115969723A - Application of salidroside in the preparation of cosmetics with anti-wrinkle and firming functions - Google Patents

Abstract

The invention relates to the field of cosmetics, and provides application of salidroside in preparation of cosmetics with skin wrinkle resisting and tightening effects. The salidroside is found for the first time to be capable of promoting the proliferation, migration and collagen synthesis of fibroblasts by regulating the expression of TGF beta 1 in macrophages, and the collagen synthesis of the fibroblasts is an important mechanism for resisting wrinkles and tightening skin. And salidroside can remove ROS in zebra fish embryos, has an antioxidant effect, and supports the functions of resisting wrinkles and tightening salidroside. Meanwhile, a rabbit skin irritation test proves that salidroside has no irritation to skin and can be applied to skin care products.

Description

Application of salidroside in the preparation of cosmetics with anti-wrinkle and firming functions

technical field

The invention relates to the field of skin care cosmetics, in particular to the application of salidroside in the preparation of cosmetics with anti-wrinkle and firming functions.

Background technique

Salidroside, or Salidroside, is a natural glycoside and one of the active ingredients of the plant Rhodiola rosea L. It has anti-fatigue, anti-hypoxia, anti-microwave radiation and other pharmacological effects. Collagen loss is one of the biggest causes of sagging skin, the appearance of wrinkles, and other signs of skin aging. Fibroblasts are the most important cell components in the dermis of the skin, capable of synthesizing and secreting collagen and elastin. The reduction in the number of fibroblasts, the change in shape, the weakening or decline of secretion and synthesis functions are closely related to skin aging, and macrophages play an important role in regulating the function of fibroblasts. In skin tissue, macrophages can release TGFβ1, and TGFβ1 acts on fibroblasts to promote the synthesis of type I and III procollagen in fibroblasts, thereby increasing the collagen content in the dermis.

The main effects of salidroside in the prior art include anti-oxidation, whitening, and anti-radiation, and other effects have not been reported yet.

Contents of the invention

In view of this, the purpose of the present invention is to provide the application of salidroside in the preparation of cosmetics with anti-wrinkle and firming functions; its application in skin anti-wrinkle and firming has been studied, and the skin stimulation effect of salidroside has been measured. .

The research on the effect and activity of salidroside on skin anti-wrinkle and firming is firstly the effect of salidroside on the expression of TGF-β in macrophages. As shown in Figure 2, 100 μM salidroside can significantly increase the expression level of TGFβ1 mRNA in the macrophage Raw264.7 cells; at the same time, 100 μM salidroside can significantly promote the release of TGFβ1 protein in Raw264.7 cells.

The research on the activity of salidroside in skin anti-wrinkle and firming includes the effect on macrophage-mediated fibroblast proliferation. As shown in Figure 3, the Raw264.7 cell supernatant treated with 100 μM salidroside can significantly promote the proliferation of fibroblast cell line L929 cells compared with the Raw264.7 cell supernatant.

The study of the activity of salidroside in skin anti-wrinkle and firming includes the effect on macrophage-mediated fibroblast migration. As shown in Figure 4, the Raw264.7 cell supernatant treated with 100 μM salidroside can significantly promote the migration of fibroblast cell line L929 cells compared with the Raw264.7 cell supernatant.

The research on the effect and activity of salidroside on skin anti-wrinkle and firming also includes the effect on macrophage-mediated fibroblast collagen synthesis. As shown in Figure 5, the Raw264.7 cell supernatant treated with 100 μM salidroside can significantly promote the mRNA expression of Collagen I and Collagen III in the fibroblast cell line L929 cells compared with the Raw264.7 cell supernatant.

The research on the activity of salidroside in skin anti-wrinkle and firming also includes the effect on ROS scavenging. As shown in Figure 6, the scavenging rate of salidroside for zebrafish embryo ROS at the test concentration of 0.05g/L was 43%, and the scavenging rate of salidroside for zebrafish embryo ROS at the test concentration of 0.01g/L was 35%. %, indicating that salidroside can significantly remove ROS in zebrafish embryos, has an antioxidant effect, and supports the functions of anti-wrinkle and firmness.

The study on the stimulating effect of salidroside on the skin applied the rabbit skin irritation model. As shown in Table 2 and Fig. 8 and Fig. 9, compared with the control group, after smearing salidroside solution, no obvious erythema and edema phenomenon occurred, and H&E section staining results showed that the rabbit skin tissue had no pathological changes (Fig. 10), so salidroside has no significant stimulating effect on rabbit skin.

The invention provides the application of salidroside as an agent for promoting collagen synthesis in the preparation of cosmetics with anti-wrinkle and firming effects.

Preferably, the cosmetic also includes auxiliary materials.

Preferably, the dosage form of the cosmetic includes cream, emulsion, water, gel, oil, powder, mud, aerosol, patch, film or nano-preparation.

The invention provides the application of salidroside in the preparation of a reagent for promoting the expression of TGFβ1 in macrophages.

The invention provides the application of salidroside in the preparation of a reagent for promoting proliferation of fibroblasts.

The invention provides the application of salidroside in the preparation of a reagent for promoting the migration of fibroblasts.

The invention provides the application of salidroside in the preparation of a reagent for promoting fibroblast collagen synthesis.

The invention provides the application of salidroside in the preparation of a reagent for removing ROS.

Preferably, the salidroside promotes fibroblast proliferation, migration and collagen synthesis through the mediation of macrophages, removes ROS in the body, and achieves anti-wrinkle and firming effects.

Compared with the prior art, the present invention has the following beneficial effects: the present invention provides the application of salidroside in the preparation of cosmetics with anti-wrinkle and firming functions; collagen synthesis of fibroblasts is an important mechanism for skin anti-wrinkle and firmness; The salidroside synthesized in the present invention can regulate the expression of TGFβ1 in macrophages, thereby promoting the proliferation, migration and collagen synthesis of fibroblasts. The results of zebrafish embryo ROS scavenging test support the anti-wrinkle and firming effects of salidroside. Rabbit skin irritation test proves that salidroside has no irritation to skin and can be used in skin care products.

Description of drawings

Figure 1 is the effect of salidroside on the viability of Raw264.7 cells;

Figure 2 is the effect of salidroside on TGFβ1 mRNA expression and protein release in Raw264.7 cells;

Figure 3 is the effect of salidroside-treated macrophage supernatant on fibroblast proliferation;

Fig. 4 is the effect of the macrophage supernatant of salidroside treatment on fibroblast migration;

Fig. 5 is the macrophage supernatant that salidroside is processed influences on the collagen synthesis in fibroblast;

Figure 6 is the level of ROS in the zebrafish embryos treated with salidroside;

Figure 7 is the detection process of salidroside on the scavenging effect of ROS in zebrafish embryos;

Fig. 8 is the representative picture of the rabbit skin irritation reaction after salidroside administration 4h;

Figure 9 is a representative picture of the skin irritation response of rabbits after salidroside administration for 72h;

Figure 10 is the H&E staining result of salidroside's irritative response to rabbit skin.

Detailed ways

The invention provides the application of salidroside as an agent for promoting collagen synthesis in the preparation of cosmetics with anti-wrinkle and firming effects.

In the present invention, the cosmetic also includes auxiliary materials, and the present invention does not specifically limit the type of the auxiliary materials, and the auxiliary materials that can be used in the field of cosmetics can be used.

In the present invention, the dosage form of the cosmetic preferably includes cream, emulsion, water, gel, oil, powder, mud, aerosol, patch, film or nano-preparation.

The present invention also provides the application of salidroside in the preparation of a reagent for promoting the expression of TGFβ1 in macrophages.

The invention also provides the application of salidroside in the preparation of reagents for promoting fibroblast proliferation, migration and collagen synthesis.

The present invention also provides the application of salidroside in the preparation of reagents for removing ROS.

In the present invention, the salidroside promotes fibroblast proliferation, migration and collagen synthesis through the mediation of macrophages, removes ROS in the body, and achieves anti-wrinkle and firming effects.

The technical solutions provided by the present invention will be described in detail below in conjunction with the examples, but they should not be interpreted as limiting the protection scope of the present invention.

Example 1

This example is the regulatory effect of salidroside on TGFβ1 gene expression in macrophages. The effect of salidroside on the viability of macrophages and the effect of salidroside on TGFβ1 mRNA expression and protein release in macrophages were measured respectively. Among them, macrophage Raw264.7 (ATCC TIB-71) cells were purchased from the Cell Bank of the Chinese Academy of Sciences. The specific implementation method is as follows:

(1) The effect of salidroside on the viability of macrophages

The cell line used was Raw264.7, the cell viability assay method was CCK8 experiment, and the salidroside action time was 48h. The results are shown in Figure 1, salidroside within 10 mM acts on Raw264.7 cells without significant cytotoxicity.

(2) Effect of salidroside on TGFβ1 mRNA expression and protein release in macrophages

The cell line used is Raw264.7. The detection method of TGFβ1mRNA expression level was qRT-PCR, and the action time of salidroside was 9h. The detection method of TGFβ1 protein release was ELISA, and the action time of salidroside was 12 hours. The results are shown in Figure 2, 100 μM salidroside can significantly increase the expression level of TGFβ1 mRNA and the release of TGFβ1 protein in Raw264.7 cells.

Example 2

This example is the effect of salidroside on the proliferation, migration and collagen synthesis of fibroblasts mediated by macrophages. The effect of salidroside on macrophage-mediated fibroblast proliferation, the effect of salidroside on macrophage-mediated fibroblast proliferation and migration, and the effect of salidroside on macrophage-mediated Effects on fibroblast collagen synthesis. Among them, macrophage Raw264.7 (ATCC TIB-71) cells and mouse fibroblast L929 were purchased from the Cell Bank of the Chinese Academy of Sciences. The specific implementation methods and results are as follows:

(1) Effect of salidroside on macrophage-mediated fibroblast proliferation

The cell lines used were macrophage Raw264.7 and fibroblast L929. The assay method of L929 cell viability is CCK8 assay. Specifically, the supernatant was collected after 100 μM salidroside was applied to Raw264.7 cells for 12 hours, and the cell viability was measured after the supernatant was applied to L929 cells for 24 hours. The results are shown in Figure 3: Raw264.7 cell supernatant treated with 100 μM salidroside (salidroside 100 μM) can significantly promote the proliferation of L929 cells compared with Raw264.7 cell supernatant (Control). Therefore, salidroside can promote the proliferation of fibroblasts by regulating macrophages.

(2) Effect of salidroside on macrophage-mediated fibroblast migration

The cell lines used were macrophage Raw264.7 and fibroblast L929. The assay method of L929 cell migration was scratch test. Specifically, the supernatant was collected after 100 μM salidroside was applied to Raw264.7 cells for 12 hours, and the cell migration was measured after the supernatant was applied to L929 cells for 24 hours. The results are shown in Figure 4: Raw264.7 cell supernatant treated with 100 μM salidroside (salidroside 100 μM) can significantly promote the migration of L929 cells compared with Raw264.7 cell supernatant (Control). Therefore, salidroside can promote the migration of fibroblasts by regulating macrophages.

(3) Effect of salidroside on collagen synthesis of fibroblasts mediated by macrophages

The cell lines used were macrophage Raw264.7 and fibroblast L929. Collagen synthesis was measured by qRT-PCR to measure the mRNA levels of Collagen I and Collagen III. Specifically, the supernatant was collected after 100 μM salidroside was applied to Raw264.7 cells for 12 hours, and the mRNA levels of Collagen I and Collagen III were measured after the supernatant was applied to L929 cells for 48 hours. The results are shown in Figure 5: compared with the Raw264.7 cell supernatant (Control), the Raw264.7 cell supernatant (salidroside 100 μM) treated with 100 μM salidroside can significantly promote the expression of Collagen I in L929 cells. mRNA expression with CollagenIII. Therefore, salidroside can promote the mRNA expression of Collagen I and Collagen III in fibroblasts by regulating macrophages.

Example 3

This example is a study on the scavenging effect of salidroside on ROS in zebrafish embryos.

The experimental organisms were zebrafish embryos. The experimental method is to transfer 48h zebrafish embryos to a 96-well plate, one fish embryo per well, set up 0.05g/L salidroside group, 0.01g/L salidroside group and blank control group, each group 24 zebrafish embryos. Place them in an incubator at 28±1°C and cultivate them until 72±1h after fertilization. Transfer to a 24-well plate, each well containing 12 fish embryos and 2mL reactive oxygen species (ROS) test solution, and then placed in a 28±1°C incubator for 2±1h. The fish embryos were placed sideways, and the fish embryos were photographed under a fluorescent stereomicroscope according to uniform photographing parameters. The specific experiment was entrusted to a third-party testing company Vitargent (International) Biotechnology Co., Ltd. to complete. The detection process is shown in Figure 7:

The calculation method of reactive oxygen species (ROS) scavenging rate is as follows:

In the above formula, T is the average value of the "average signal intensity" of fish embryo ROS in the test substance treatment group, and C is the average value of the "average signal intensity" of fish embryo ROS in the blank control group. A two-tailed T-test was performed on the ROS signal intensity of fish embryos in the test substance treatment group and the ROS signal intensity of the blank control group, and the p value was obtained.

Example 4

This example is a study on the stimulating effect of salidroside on rabbit skin.

The experimental animals were clean grade New Zealand rabbits (2-2.2 kg), which were purchased from the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine (Shanghai Chedun Experimental Animal Breeding Farm Co., Ltd.). The preparation method of the salidroside solution is as follows: 300 mg of salidroside drug powder is weighed and dissolved in 300 ml of physiological saline to obtain a 100 mg/ml salidroside solution. The hair on both sides of the rabbit's back spine was shaved without damaging the skin. Dip an appropriate amount of salidroside drug solution with a cotton swab and apply it directly on the skin, apply gauze on it after application, and then fix it with non-irritating adhesive tape. Physiological saline was applied to the shaved skin on the other side as a negative control. Under natural light, the skin reaction was observed with the naked eye after 4h, 24h, 48h, and 72h of application, and the skin irritation reaction was scored according to Table 1. After the experiment, the total score (stimulation index) of each rabbit was calculated to determine the skin irritation intensity.

Table 1 Skin irritation reaction scoring method

The experimental results are shown in Table 2 and Figure 8 and Figure 9. Compared with the control group, no obvious erythema and edema appeared after the salidroside solution was applied, and the results of H&E section staining showed that the rabbit skin tissue had no pathological changes (FIG. 10), so salidroside has no significant stimulating effect on rabbit skin.

Table 2 The scoring of salidroside on rabbit skin irritation

It can be seen from the above examples that the salidroside synthesized in the present invention can regulate the expression of TGFβ1 in macrophages, thereby promoting the proliferation, migration and collagen synthesis of fibroblasts. Rabbit skin irritation test proves that salidroside has no irritation to skin and can be used in skin care products.

The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims (7)

1. The application of salidroside in the preparation of cosmetics with anti-wrinkle and firming effects as a collagen synthesis agent. 2. application according to claim 1, is characterized in that, described cosmetic also comprises auxiliary material. 3. application according to claim 1, is characterized in that, the dosage form of described cosmetic comprises cream, emulsion, water, gel, oil, powder, mud, aerosol, patch, Films or nano-formulations. 4. The application of salidroside in the preparation of a reagent for promoting the expression of TGFβ1 in macrophages. 5. Application of salidroside in the preparation of reagents for promoting proliferation, migration and collagen synthesis of fibroblasts. 6. The application according to claim 1, characterized in that the salidroside is mediated by macrophages to promote fibroblast proliferation, migration and collagen synthesis. 7. The application according to claim 1, characterized in that the salidroside can scavenge ROS and has anti-oxidation effect.

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