Application of ginsenoside Rh1 in preparation of product for relieving skin photoaging

Abstract

The invention belongs to the technical field of biology, and particularly relates to application of ginsenoside Rh1 in preparation of a product for relieving skin photoaging. It is found for the first time that the rare ginsenoside Rh1 efficiently removes ROS caused by UV and inhibits cell aging by activating a Sirt3 / Nrf2 / HO-1 pathway, and a brand new strategy is provided for developing a natural anti-photoaging preparation with repair and protection functions.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to application of ginsenoside Rh1 in preparation of a product for relieving skin photoaging. BACKGROUND

[0002] Skin aging is a complex biological process mediated by endogenous metabolic disorders and exogenous environmental stress. Endogenous aging is mainly driven by genetic factors and cell metabolic function decline, manifested as collagen loss and elastic fiber rupture. Exogenous aging is mainly photoaging, and the core cause is oxidative stress induced by ultraviolet radiation. Ultraviolet rays are divided into UVA with a wavelength of 320nm-400nm, UVB with a wavelength of 280nm-320nm and UVC with a wavelength of 200nm-280nm, among which UVA and UVB are the main causes of photoaging. UVB can directly damage epidermal cell DNA due to high energy and strong penetration, and activate the excessive generation of active oxygen, ultimately accelerating the process of skin photoaging.

[0003] Current intervention strategies for photoaging mainly include chemical antioxidants, physical barrier protection and instrument treatment. Chemical antioxidants such as vitamin C, vitamin E and coenzyme Q10 can reduce oxidative damage by neutralizing free radicals, but high concentrations for long-term use can easily cause skin irritation, photosensitivity and other problems. Physical sunscreen agents such as titanium dioxide and zinc oxide can reflect or absorb ultraviolet rays, but their nanoparticles can penetrate the skin and cause inflammation, and cannot repair the oxidative damage that has already been formed. Laser, radio frequency and other instrument treatments can improve skin texture in the short term, but have limitations such as high treatment cost, long recovery period and risk of pigmentation. Therefore, the development of natural, safe and functional anti-photoaging ingredients that can repair and protect has become a research hotspot.

[0004] As the main active ingredient of traditional Chinese medicine ginseng, ginsenoside has attracted attention in recent years due to its multi-target anti-aging properties. Among them, the rare ginsenoside Rh1, i.e. 20(S)-protopanaxatriol, is a metabolite of protopanaxadiol-type saponins, and has better bioavailability and antioxidant activity. Studies have shown that ginsenoside Rh1 can reduce liver damage by inhibiting the TAK1 / STAT3 inflammatory pathway, and improve vascular endothelial oxidative stress by activating the Nrf2 / HO-1 signaling axis. In the field of skin, some ginsenosides such as Rg3 and Rb1 have been confirmed to inhibit the expression of metalloproteinase-1 (MMP-1) and promote collagen synthesis. Therefore, further research on ginsenoside Rh1 is needed to provide more applications. SUMMARY

[0005] To solve the above technical problems, the present application provides application of ginsenoside Rh1 in preparation of a product for relieving skin photoaging.

[0006] The technical scheme of the present application is as follows.

[0007] Application of ginsenoside Rh1 in preparation of products for relieving skin photoaging.

[0008] The present application first discovers that ginsenoside Rh1 can effectively remove ROS caused by UV and inhibit cell aging by activating the Sirt3 / Nrf2 / HO-1 cascade signal pathway, thereby providing a new strategy for developing natural anti-photoaging products with repair and protection functions. In a UV-induced mouse photoaging model, ginsenoside Rh1 can reduce epidermal thickening, relieve erythema and skin moisture loss, and increase collagen fibers in the dermis. By adjusting the Bcl-2 / Bax protein ratio, cell apoptosis is inhibited; in a UVB-induced human skin fibroblast cell model, ginsenoside Rh1 significantly delays cell aging, reduces the proportion of SA-β-gal positive cells, and thus reduces HDF cell oxidative stress. The study found that ginsenoside Rh1 can activate mitochondrial deacetylase Sirt3, promote the nuclear translocation of nuclear factor Nrf2, up-regulate the expression of downstream heme oxygenase-1, and inhibit UVB-induced skin oxidative stress and aging.

[0009] In another preferred embodiment, the product is a skin external preparation with a ginsenoside Rh1 concentration of 1 μM to 100 μM.

[0010] In another preferred embodiment, the product is a skin external preparation with a ginsenoside Rh1 concentration of 12.5 μM to 50 μM.

[0011] In another preferred embodiment, the ginsenoside Rh1 is the only effective ingredient in the skin external preparation.

[0012] In another preferred embodiment, the skin external preparation is any one of a cream, a gel, a solution, or a nano-liposome.

[0013] In another preferred embodiment, the solvent in the solution is a dimethyl sulfoxide solution with a mass percentage of 0.01% to 0.1% or an ethanol solution with a mass percentage of 0.01% to 0.1%. The dimethyl sulfoxide solution is obtained by diluting dimethyl sulfoxide with physiological saline.

[0014] In another preferred embodiment, the skin photoaging refers to photoaging caused by ultraviolet rays.

[0015] In another preferred embodiment, the photoaging caused by ultraviolet rays refers to skin oxidative stress and aging caused by ultraviolet rays.

[0016] Compared with the prior art, the present application has the following beneficial effects.

[0017] The present invention has demonstrated through experiments that ginsenoside Rh1 can reduce epidermal thickening, alleviate erythema and skin water loss, and increase dermal collagen fibers in a UV-induced mouse photoaging model. It also inhibits apoptosis by regulating the Bcl-2 / Bax protein ratio. Furthermore, ginsenoside Rh1 can reduce UVB-induced ROS production and photoaging in HDF cells. Further experiments revealed that ginsenoside Rh1 inhibits UVB-induced skin oxidative stress and aging by activating the mitochondrial deacetylase Sirt3, promoting the nuclear translocation of the nuclear factor Nrf2, and upregulating downstream heme oxygenase-1 expression. This suggests that the rare ginsenoside Rh1 can be used to prepare products that target SIRT3 to combat skin oxidative stress and alleviate skin photoaging, providing a new application for ginsenoside Rh1. Furthermore, experiments have shown that ginsenoside Rh1 is safe, reliable, and has significant efficacy without toxic side effects.

[0018] The present invention solves the technical problem of side effects of existing chemically synthesized drugs and devices for treating skin photoaging, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Figure 1 is a graph showing the effect of ginsenoside Rh1 on alleviating skin aging in vivo; A is a representative image of the changes in the back skin morphology of each group of mice after 8 weeks of ultraviolet irradiation, in which Control represents the UV-negative 0.01wt% DMSO solution + saline group, UV represents the UV-positive 0.01wt% DMSO + saline group, Rh1-25μM represents the Rh1 low-dose group, Rh1-50μM represents the Rh1 high-dose group, and Retino-A group; B is a representative image of epidermal thickness stained with HE, with a scale bar of 50μm; C is a graph showing the erythema volume and moisture content of mice; D is a graph showing the moisture content of the back skin of mice, in which - represents no addition, + represents the addition of the corresponding substance, 25 represents the addition of 25μM Rh1, and 50 represents the addition of 50μM Rh1; E is a graph showing the results of Western blot detection of apoptotic proteins; F is a graph showing the relative protein expression of Bcl-2 / Bax; n=6, ns, no significant difference, * P <0.05,** P <0.01,*** P <0.001; in the figure, 1 represents Control, 2 represents UV, 3 represents UV+Rh1-25 μM, 4 represents UV+Rh1-50 μM, and 5 represents UV+Retino-A; DMSO solution represents dimethyl sulfoxide solution.

[0020] Figure 2Figure for results of ginsenoside Rh1 protecting HDF cells from UVB-induced cell senescence; wherein, A is a figure for SA-β-gal positive staining indicating senescent cells, scale bar is 50 μm; B is a figure for measuring ROS content by flow cytometry; C is a quantitative analysis result of B; D is a figure for measuring ROS content by fluorescence, scale bar is 10 μm, in the figure, DAPI is a nuclear dye, DCFH-DA is a fluorescent dye, verifying whether ROS is produced, the brighter the fluorescence, the more ROS is generated; E is a quantitative analysis result of D; F is a figure for Western blot detecting apoptosis proteins; G is a relative protein expression amount of Bcl-2 / Bax; H is a figure for Western blot detecting apoptosis proteins; I is a relative protein expression amount of Bcl-2 / Bax, n=3; ns is no significant difference, P <0.05,** P <0.01,*** P <0.001; in the figure, 1 represents Control, 2 represents UVB, 3 represents UVB+Rh1-12.5 μM, 4 represents UVB+Rh1-25 μM, 5 represents UVB+Rh1-50 μM.

[0021] Figure 3 Figure for analysis of ginsenoside Rh1 enhancing expression of Sirt3, wherein, A is a figure for Western blot detecting Sirt3 protein; B is a figure for relative protein expression amount of Sirt3 / β-actin; C is a figure for cell immunostaining of Sirt3, wherein, the position of the nucleus is marked by DAPI staining, scale bar is 10 μm; D is a figure for analysis of average fluorescence intensity of Sirt3 protein; E is a figure for Western blot detecting expression of Nrf2 protein in the nucleus; F is a figure for relative protein expression amount of Nrf2 / Lamin B1; G is a figure for Western blot detecting expression of HO-1 protein; H is a figure for relative protein expression amount of HO-1 / β-actin, n=3, ns is no significant difference, P <0.05,** P <0.01,*** P <0.001; in the figure, 1 represents Control, 2 represents UVB, 3 represents UVB+Rh1-12.5 μM, 4 represents UVB+Rh1-25 μM, 5 represents UVB+Rh1-50 μM.

[0022] Figure 4Figure 1 shows the results of ginsenoside Rh1 protecting HDF cells from UV-induced cellular senescence via the Sirt3 / Nrf2 / HO-1 pathway. A shows the expression of Sirt3, HO-1, and nuclear Nrf2 proteins by Western blot; B shows the relative protein expression of HO-1 / β-actin; C shows the relative protein expression of Sirt3 / β-actin; and D shows the relative protein expression of Nrf2 / Lamin B1. n = 3, ns, no significant difference, * P <0.05,** P <0.01,*** P <0.001; in the figure, 1 represents Control, 2 represents UVB, 3 represents UVB+Rh1-50μM, 4 represents UVB+Rh1-50μM+Si-Sirt3, and 5 represents UVB+Rh1-50μM+Si-NC.

[0023] Figure 5 Figure 1 shows the results of Nrf2 nuclear translocation. A shows the results of Nrf2 immunostaining in cells, with the nucleus marked by DAPI staining. The scale bar is 10 μm. B shows the analysis of Nrf2 fluorescence intensity. C shows the analysis of Nrf2 nuclear fluorescence intensity. n = 3, ns, no significant difference, * P <0.05,** P <0.01,*** P <0.001; in the figure, 1 represents Control, 2 represents UVB, 3 represents UVB+Rh1-50μM, 4 represents UVB+Rh1-50μM+Si-Sirt3, and 5 represents UVB+Rh1-50μM+Si-NC.

[0024] Figure 6 The figures are the results of ROS content and positive SA-β-gal staining indicating senescent cells; A is the result of flow cytometry measurement of ROS content; B is the result of fluorescence quantitative analysis of A; C is the result of positive SA-β-gal staining indicating senescent cells, scale bar is 50 μm, n=3; ns, no significant difference, *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0025] In order to more clearly understand the above-mentioned objects, features and advantages of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be noted that, in the absence of conflict, the embodiments of the present application and the features therein can be combined with each other.

[0026] Silencing regulator protein 3 is marked as Sirt3; hemoglobin oxygenase-1 is marked as HO-1; active oxygen is marked as ROS.

[0027] Ginsenoside Rh1 is provided by Ginseng Research Institute of Jilin University, the molecular weight is 638.88, the purity is greater than or equal to 98%, the molecular formula is C 36 H 62 O9, PubChem CID is 12855920, CAS number is 63223-86-9, and is referred to as Rh1 in the following examples. 7-week-old male BALB / c mice are purchased from Beijing Huafukang Biotechnology Co., Ltd.; 0.05g / 100g all-trans retinoic acid is purchased from MENARINI company. 0.05g / 100g retinoic acid is referred to as Retino-A in the following examples.

[0028] The role and mechanism of ginsenoside Rh1 in skin photoaging have not been elucidated, especially whether it plays an effect by regulating Sirt3 / Nrf2 / HO-1. Sirt3, as a key mitochondrial deacetylase, plays a core role in delaying aging by regulating antioxidant enzyme activity and maintaining metabolic homeostasis. UVB radiation can significantly down-regulate Sirt3 expression, leading to mitochondrial ROS accumulation and cell apoptosis. Activation of Sirt3 can promote the nuclear translocation of transcription factor Nrf2 and the expression of downstream antioxidant enzymes such as HO-1, forming a cascade defense network. However, there is no study to reveal the molecular mechanism of natural compounds targeting Sirt3 / Nrf2 / HO-1 pathway to improve skin photoaging.

[0029] Therefore, there is a lack of natural anti-photoaging components targeting Sirt3 signaling axis in the prior art, and traditional antioxidants have safety and efficacy limitations. The present application first discovers that the rare ginsenoside Rh1 can effectively remove ROS caused by UV and inhibit cell senescence by activating the Sirt3 / Nrf2 / HO-1 pathway, providing a new strategy for developing natural anti-photoaging preparations with repair and protection functions.

[0030] The application of ginsenoside Rh1 in preparing products for relieving skin photoaging is specifically described below.

[0031] Example 1: In vivo anti-photoaging of ginsenoside Rh1.

[0032] 7-week-old male BALB / c mice are selected for the experiment and are raised in a standardized experimental animal room with a temperature of 24±2℃, humidity of 60%±5%, and a 12h light-dark cycle, and are allowed to freely eat and drink. All animal experiments are approved by the Animal Ethics Committee of West China Hospital of Sichuan University, with approval number: 20241230001. The following dimethyl sulfoxide solution is referred to as DMSO solution.

[0033] The mice were randomly divided into 5 groups, 6 in each group, which were UV negative, 0.01% DMSO solution + physiological saline group, UV positive, 0.01% DMSO solution + physiological saline group, Rh1 low-dose group, Rh1 high-dose group and Retino-A group. Among them, 0.01% DMSO solution was obtained by diluting DMSO with physiological saline; the concentration of Rh1 in the Rh1 low-dose group was 25 μM, and the concentration of Rh1 in the Rh1 high-dose group was 50 μM. Among them, the physiological saline is a 0.9 wt% sodium chloride solution.

[0034] 30 min before UV irradiation, 150 μL of the above-mentioned 5 groups of solutions were respectively applied to the back skin of each mouse to evaluate the inhibitory effect of Rh1 on photoaging. UV irradiation was performed at a distance of 30 cm from the mouse skin, 5 times a week, for a total of 8 weeks, and the irradiation intensity is shown in Table 1. In the ninth week, the mice were anesthetized with tri- bromoethanol, and the color difference, moisture content were determined, and then the skin samples were taken for Western blot experiment and HE staining.

[0035] Table 1 UV irradiation intensity .

[0036] The experimental results are shown in Figure 1 . As shown in A of Figure 1 , after 8 weeks of UV irradiation, the mouse skin showed typical large area lesions. In contrast, the skin of the control mice during the experiment remained healthy, with no obvious wrinkles. After 8 weeks of UV irradiation, the UV group mice showed skin damage, manifested as coarse wrinkles, erythema, thickening and leather-like texture, indicating that the skin aging model was successfully established. On the contrary, the skin damage of the irradiated mice receiving the Rh1 low-dose group, the Rh1 high-dose group and the Retino-A group was significantly alleviated, with only a small amount of superficial wrinkles and mild erythema, and no obvious skin lesions were observed.

[0037] To further evaluate the anti-aging effect of Rh1, histological analysis of skin tissue sections was performed using H&E staining. The epidermis of healthy skin is composed of multiple layers of squamous cells covered by a thin layer of stratum corneum, and the dermis layer is mainly composed of dense connective tissue, including collagen fibers and elastic fibers, etc. Compared with the normal control group, the epidermis layer of the UV irradiation group mice increased significantly, and the collagen fibers in the dermis layer decreased significantly and arranged in disorder. However, the use of Rh1 low-dose group, Rh1 high-dose group and Retino-A group can reduce the thickness of epidermis layer and increase the collagen fibers in dermis layer, as shown in Figure 1B. Next, the skin erythema and skin moisture content were evaluated using a colorimeter and a skin moisture meter. These analyses showed that UV radiation increased the amount of skin erythema and decreased the moisture content. In contrast, the amount of skin erythema decreased and the moisture content increased in the skin of mice treated with the Rh1 low dose group, the Rh1 high dose group, as shown in C and D of FIG. 1. Figure 1 Excessive apoptosis is the culprit of skin photoaging. Therefore, the expression of the anti-apoptotic protein Bcl-2 and the pro-apoptotic protein Bax in the skin tissue of mice was detected. The apoptosis of the skin of mice in the UV group increased compared with the control group. In contrast, the apoptosis of mice treated with the Rh1 low dose group, the Rh1 high dose group, or the Retino-A group decreased as shown in E and F of FIG. 1. Figure 1 The above results show that Rh1 can prevent UV-induced skin aging in vivo.

[0038] Example 2: Ginsenoside Rh1 delays HDF cell aging in vitro.

[0039] (1) HDF cells were cultured in DMEM F12 medium containing 1 wt% penicillin-streptomycin and 10 w% fetal bovine serum, and the culture conditions were 37°C, 5% CO2 in air in a constant temperature incubator. When the cells reached about 80% confluence, they were passaged at a density of 5 x 103 cells per well in a 96-well plate. When the cells reached about 70% confluence, they were intervened, and different concentrations of Rh1 were added to the 96-well plate to detect their cytotoxicity. The ultraviolet lamp was preheated for 10 min before UVB irradiation, the culture medium was removed, and the cells were washed twice with PBS. A layer of PBS was added on the well plate to cover the cells, and then different concentrations of UVB irradiation were performed to find the most UVB intensity. After the intervention, 10 μL of CCK8 reagent was added to each well, and incubated at 37°C for 2 h. The absorbance value OD 450 .

[0040] Cell viability (%) = [(experimental group OD value - blank group OD value) / (negative control group OD value - blank group OD value)] x 100%.

[0041] The experimental results are shown in Tables 2 and 3. First, the cytotoxicity of Rh1 on HDF was evaluated, and it was found that when the concentration of Rh1 was higher than 50 µM, the viability of HDF would decrease, as shown in Table 2. Therefore, in the subsequent experiments, three concentrations of 12.5 µM, 25 µM and 50 µM were selected. The cells were exposed to 0 mJ / cm 2 , 20 mJ / cm 2 , 40 mJ / cm 2 , 60 mJ / cm 2 , 80 mJ / cm 2 and 100 mJ / cm 2in UVB and the cell viability was analyzed to determine the intensity of UVB used in the experiment. At 40 mJ / cm 2 , the HDF cell viability was about 75% as shown in Table 3. Therefore, the intensity of 40 mJ / cm 2 was chosen for the subsequent experiments.

[0042] Table 2 Cytotoxicity of different concentrations of Rh1 .

[0043] Table 3 Cytotoxicity of different intensities of UVB .

[0044] (2) When the cells reached about 80% confluence, they were passaged, and when the cells reached 70% confluence, the HDF cells were pretreated with Rh1 at concentrations of 12.5 µM, 25 µM, and 50 µM for 24 h, respectively. The ultraviolet lamp was preheated for 10 min before UVB irradiation. The culture medium was removed, and the cells were washed twice with PBS. A thin layer of PBS was added to the well plate to cover the cells, and UVB irradiation was performed at a distance of 20 cm from the cells at an intensity of 40 mJ / cm 2 . After irradiation, the PBS was removed, and DMEM F12 medium was added to the cells for continuous culture. After 24 h, the β-galactosidase experiment was performed to detect senescent cells, flow cytometry and fluorescence experiments were performed to detect the ROS content in HDF cells, and Western blot experiment was performed to detect the expression of apoptosis proteins.

[0045] The experimental results are shown in Figure 2 . To evaluate the effect of Rh1 treatment on UVB-induced cell aging, SA-β-Gal staining was performed after UVB and Rh1 treatment. The results showed that the number of SA-β-gal positive cells increased significantly after UVB irradiation and appeared blue. In contrast, the number of the above-mentioned cells in the Rh1 treatment group decreased significantly depending on the dose, as shown in A of Figure 2 . During skin aging, the accumulation of oxidative products and the dysfunction of organelles induced by oxidative stress disrupt cellular metabolic homeostasis, ultimately leading to cell apoptosis. As shown in B-E of Figure 2 , compared with the control group, the ROS in HDF cells increased after UVB irradiation, while the ROS decreased significantly after Rh1 treatment. The expression of apoptosis-related proteins Bcl-2 and Bax was then detected, and the results showed that, compared with the control group, the expression of anti-apoptotic protein Bcl-2 decreased, the expression of pro-apoptotic protein Bax increased, and Bcl-2 / Bax decreased after UVB irradiation, while apoptosis decreased significantly after Rh1 treatment as shown in Figure 2After determining these effects of Rh1, it was then determined whether Rh1 could reduce the occurrence of apoptosis by preventing the production of ROS. For this, the ROS inhibitor NAC was used, which has a significant free radical scavenging activity and is an effective antioxidant that can scavenge ROS produced in a series of different enzymatic reactions, thereby inhibiting oxidative damage. The results showed that NAC significantly inhibited the occurrence of apoptosis, with an inhibitory effect comparable to that of Rh1, indicating that in protecting cells from oxidative damage, Rh1 mainly reduces apoptosis by reducing the production of ROS. The above results show that Rh1 can prevent UVB-induced skin aging in vitro.

[0046] Example 3: Ginsenoside Rh1 enhances Sirt3 expression, promotes nuclear translocation of Nrf2 and expression of HO-1.

[0047] When the cells reached about 80% confluence, they were passaged, and when the cells reached 70% confluence, the HDF cells were pretreated with Rh1 at concentrations of 12.5 µM, 25 µM and 50 µM for 24 h, respectively. The ultraviolet lamp was preheated for 10 min before UVB irradiation. The culture medium was removed, and the cells were washed twice with PBS. A thin layer of PBS was added to the well plate to cover the cells, and UVB irradiation was performed at a distance of 20 cm from the cells and an irradiation intensity of 40 mJ / cm2. After irradiation, the PBS was removed, and DMEM F12 medium was added to the cells for continued culture. After 24 h, Western blotting was performed to detect the expression of Sirt3, Nrf2 and HO-1. 2

[0048] The experimental results are shown in Figure 3 . First, Western blotting was performed to detect whether Rh1 could change the expression of Sirt3. As shown in Figure 3 A and B, compared with the control group, UVB irradiation significantly reduced the expression of Sirt3 protein, and the expression of Sirt3 was significantly restored after treatment with 50 µM Rh1. Although 12.5 µM Rh1 and 25 µM Rh1 also increased the expression of Sirt3 to a certain extent, it did not reach statistical significance. At the same time, the expression of Sirt3 fluorescent protein was also detected by immunofluorescence experiment, and the results showed that Rh1 could significantly inhibit the weakening of Sirt3 fluorescence intensity caused by UVB irradiation, as shown in Figure 3 C and D. Next, the expression of Nrf2 protein in the nucleus was detected, and the results showed that Rh1 significantly increased the nuclear translocation of Nrf2, as shown in Figure 3 E and F. In addition, Rh1 also significantly increased the expression of HO-1, as shown in Figure 3 G and H. The above results show that RH1 enhances Sirt3 expression, promotes nuclear translocation of Nrf2 and expression of HO-1.​

[0049] Example 4: Ginsenoside Rhl protects HDF cells from aging by activating Sirt3 / Nrf2 / HO-1 pathway.

[0050] Example 4: Ginsenoside Rhl protects HDF cells from aging by activating Sirt3 / Nrf2 / HO-1 pathway. Figures 4-6

[0051] Firstly, the Sirt3 knockdown efficiency was found by Western blot as shown in Table 4.

[0052] Table 4 Sirt3 knockdown efficiency .

[0053] Then, after establishing the aging model by UVB radiation, it was found that the expression of Sirt3 protein was reduced in the UVB radiation group compared with the Control group, and Rhl treatment could restore its expression, and this effect was blocked by Sirt3 silencing, while the si-NC treatment group continued to maintain the restoring effect of Rhl as shown in A and B of Figure 4 Figure 4

[0054] In addition, it was found by immunofluorescence experiment that compared with the UVB radiation group, Rhl increased the expression of total fluorescent protein and nuclear fluorescent protein of Nrf2, suggesting the nuclear translocation of Nrf2. Sirt3 silencing inhibited the expression of total fluorescent protein and nuclear fluorescent protein, suggesting the inhibition of Nrf2 nuclear translocation, while the si-NC treatment group continued to maintain the restoring effect of Rhl as shown in A, B and C of Figure 5

[0055] Next, it was found by flow cytometry that Rhl significantly inhibited the production of ROS in HDF cells. Sirt3 silencing increased the production of ROS, inhibited the antioxidant effect of Rhl, while the si-NC treatment group continued to maintain the restoring effect of Rhl as shown in Figure 6 ​​​​A and B in FIG. 6C. Finally, it was found by SA-β-Gal staining that Rh1 significantly inhibited SA-β-gal positive cells, Sirt3 silencing increased the generation of SA-β-gal positive cells, reduced the anti-aging effect of Rh1, and si-NC treatment group continued to maintain the restoration effect of Rh1 as Figure 6 C in FIG. 6C. In summary, these data suggest that Sirt3 plays a key role in Rh1 improving skin aging, and Rh1 protects HDF cells from UVB-induced cell aging through the Sirt3 / Nrf2 / HO-1 pathway.

[0056] The above embodiments are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above embodiments, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and are included in the protection scope of the present application.

Claims

1. Application of ginsenoside Rh1 in the preparation of products for alleviating skin photoaging.

2. The use of ginsenoside Rh1 according to claim 1 in preparing a product for alleviating skin photoaging, characterized in that: The product is a skin external preparation with a ginsenoside Rh1 concentration of 1 μM to 100 μM.

3. The use of ginsenoside Rh1 according to claim 2 in the preparation of a product for alleviating skin photoaging, characterized in that: The product is a skin external preparation with a ginsenoside Rh1 concentration of 12.5 μM to 50 μM.

4. The use of ginsenoside Rh1 according to claim 3 in preparing a product for alleviating skin photoaging, characterized in that: The ginsenoside Rh1 is the only active ingredient in the skin external preparation.

5. The use of ginsenoside Rh1 according to claim 3 in preparing a product for alleviating skin photoaging, characterized in that: The skin external preparation is any one of a cream, a gel, a solution or a nanoliposome.

6. The use of ginsenoside Rh1 according to claim 5 in preparing a product for alleviating skin photoaging, characterized in that: The solvent in the solution is a dimethyl sulfoxide solution with a mass percentage of 0.01% to 0.1% or an ethanol solution with a mass percentage of 0.01% to 0.1%; The dimethyl sulfoxide solution is obtained by diluting dimethyl sulfoxide with physiological saline.

7. The use of ginsenoside Rh1 according to claim 1 in preparing a product for alleviating skin photoaging, characterized in that: The skin photoaging refers to light aging caused by ultraviolet rays.

8. The use of ginsenoside Rh1 according to claim 7 in preparing a product for alleviating skin photoaging, characterized in that: The ultraviolet-induced photoaging refers to skin oxidative stress and aging caused by ultraviolet rays.

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