Application of ginsenoside Rc in preparation of products for resisting skin high temperature and light damage

Abstract

The invention discloses application of ginsenoside Rc in preparation of a product for resisting skin high temperature and light damage. The skin high temperature and light damage resistance is reflected in the following aspects: resisting inflammation induced by high temperature; the inflammation induced by high temperature and ultraviolet light products FICZ is resisted. Research results show that ginsenoside Rc can significantly reduce the induction effect of FICZ on IL-6 and IL-8 in a high-temperature environment, and provides a potential protection mechanism for resisting inflammatory reactions aggravated by the high-temperature condition. The discovery not only provides a theoretical basis for application of ginsenoside Rc in defense of skin inflammation induced by environmental factors, but also reveals a potential anti-inflammatory mechanism of ginsenoside Rc on a cellular level, and provides a scientific basis for subsequent application and development in the field of skin care products.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to the application of ginsenoside Rc in the preparation of products that resist skin high temperature and light damage. Background Art

[0002] Ginsenoside Rc, a major saponin active ingredient in Panax ginseng, has attracted widespread attention due to its significant biological activity in the biomedical field, especially in skin health management. [1] Specifically, the effects of ginsenoside Rc in anti-inflammatory, promoting skin healing, anti-oxidation and enhancing skin barrier function have made it an important research subject in the fields of skin science research and skin care product development. [2-4] .

[0003] As global climate change, particularly global warming, poses increasing challenges to human health, the skin, as the body's largest external barrier, is particularly sensitive to changes in the external environment. Skin cells face challenges ranging from minor temperature fluctuations to significant temperature increases, significantly impacting their normal function and survival. While ginsenoside Rc has achieved a certain degree of application in the health sector, particularly in skincare, research and utilization of ginsenoside Rc in its protection against temperature-mediated effects remains underdeveloped. Existing technologies have not fully explored the potential of ginsenoside Rc for addressing skin damage in high-temperature environments.

[0004] References:

[0005] 1. Lim, TG, Lee, CMC, Dong, Z., Lee, K. (2015). Ginsenosides and their metabolites: a review of their pharmacological activities in the skin. Archives of Dermatological Research, 307, 397-403.

[0006] 2. Paul, S., Shin, HS, Kang, SC (2012). Inhibition of inflammations and macrophage activation by ginsenoside-Re isolated from Korean ginseng (Panaxginseng CAMeyer). Food and Chemical Toxicology, 50 (5), 1354-61.

[0007] 3.Yu,T.,Rhee,MH,Lee,J.,Kim,SH,Yang,Y.,Kim,HG,...Cho,JY(2016).Ginsenoside Rc from Korean Red Ginseng(Panax ginseng CAMeyer)AttenuatesInflammatory Symptoms of Gastritis,Hepatitis and Arthritis.The AmericanJournal of Chinese Medicine,44(3),595-615.

[0008] 4. Kim, E., Kim, W. (2018). An Insight into Ginsenoside Metabolite CompoundK as a Potential Tool for Skin Disorder. Evidence-based Complementary and Alternative Medicine: eCAM, 2018. Summary of the Invention

[0009] One object of the present invention is to provide a new use of ginsenoside Rc.

[0010] The new use of ginsenoside Rc provided by the present invention is its use in preparing products that resist skin high temperature and light damage.

[0011] The resistance to skin high temperature and light damage is embodied in the following aspects:

[0012] (1) Resist inflammation induced by high temperature;

[0013] (2) Resist inflammation induced by high temperature and ultraviolet light product FICZ.

[0014] The inflammation is caused by high temperature inducing skin cells to secrete inflammatory factors IL-6 and / or IL-8.

[0015] The inflammation is caused by high temperature and ultraviolet light product FICZ inducing skin cells to secrete inflammatory factors IL-6 and / or IL-8.

[0016] Under high temperature conditions, the UVB photoproduct 6-Formylindolo[3,2-b]carbazole (FICZ) has a significant enhancing effect on the expression of inflammatory factors such as interleukin-6 (IL-6) and interleukin-8 (IL-8) in skin keratinocytes during ultraviolet irradiation.

[0017] The method of resisting inflammation induced by high temperature or the method of resisting inflammation induced by high temperature and ultraviolet light product FICZ is to inhibit the gene expression level and / or protein expression level of inflammatory factors IL-6 and IL-8 by ginsenoside Rc.

[0018] The high temperature may be in the range of 37-38.5°C.

[0019] The above-mentioned product of the present invention can be a medicine and / or a cosmetic.

[0020] The structural formula of ginsenoside Rc is shown in (Formula I):

[0021]

[0022] Another object of the present invention is to provide a skin care product that resists skin damage caused by high temperature.

[0023] The skin care product for resisting skin damage from high temperature provided by the present invention comprises ginsenoside Rc as an active ingredient.

[0024] In the research process of the present invention, HaCaT human epidermal keratinocytes were used as a research model to reveal the inflammatory response of cells to light damage under elevated temperature conditions, and to evaluate the regulatory effect of ginsenoside Rc on the inflammatory factors IL-6 and IL-8 under elevated temperature conditions. The inventors found that under simulated elevated temperature conditions, after increasing the cell culture temperature by 1.5°C, the UVB photoproduct 6-Formylindolo[3,2-b]carbazole (FICZ) significantly enhanced the expression of inflammatory factors such as interleukin-6 (IL-6) and interleukin-8 (IL-8) in skin keratinocytes. This discovery reveals the association between heat and IL-6 and IL-8, key cell signaling molecules in inflammatory responses, and also emphasizes that high temperature environments may exacerbate the risk of UVB-induced skin damage.

[0025] In this study, we further explored the effects of ginsenoside Rc on inflammatory responses under these temperature conditions. The results showed that ginsenoside Rc significantly reduced the induction of IL-6 and IL-8 by FICZ under high-temperature conditions, providing a potential protective mechanism against inflammatory responses exacerbated by high temperatures. This finding not only provides a theoretical basis for the application of ginsenoside Rc in protecting against environmentally induced skin inflammation but also reveals its potential anti-inflammatory mechanism at the cellular level, providing a scientific basis for its subsequent application and development in skincare products.

[0026] In summary, the study of ginsenoside Rc not only enriches our understanding of its biological activity but also provides new strategies for the development of novel skincare products to address the limitations of existing technologies in protecting cells in high-temperature environments. This has important practical implications for skin health management, particularly in the context of global climate change. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The effects of ginsenoside Rc and FICZ on the activity of HaCaT cells treated with elevated temperature;

[0028] Figure 2 Ginsenoside Rc reduces the gene expression levels of IL-6 and IL-8 in HaCaT cells under elevated temperature conditions;

[0029] Figure 3 Ginsenoside Rc reduces the protein secretion levels of IL-6 and IL-8 in HaCaT cells under elevated temperature conditions; DETAILED DESCRIPTION

[0030] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0031] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0032] The reagents and instruments used in the following examples are as follows:

[0033] Keratinocytes (HaCaT cells) were purchased from the Cell Resource Center of the Chinese Academy of Medical Sciences.

[0034] Ginsenoside Rc reagent was purchased from Solarbio (Shanghai)

[0035] CCK8 kit was purchased from Sangon Biotech (Shanghai)

[0036] IL-6 and IL-8 ELISA kits were purchased from Solarbio (Shanghai)

[0037] Gene RNA was purchased from Thermo Fisher Scientific (USA)

[0038] cDNA synthesis kit was purchased from Thermo Fisher Scientific (USA)

[0039] qRT-PCR Master Mix Gibco (USA)

[0040] NanoDrop 2000 / 2000c spectrophotometer (Thermos Fisher Scientific)

[0041] mRNA TM 6Flex Real-Time PCR System (Thermo Fisher Scientific)

[0042] Example 1: Effects of ginsenoside Rc and FICZ on the activity of HaCaT cells treated with elevated temperatures

[0043] HaCaT cells were cultured at a rate of 1 × 10 5 Cells were inoculated into 96-well plates at a concentration of 100 μL / mL, and 100 μL of cell suspension was added to each well. The cells adhered to the well wall during overnight culture. Subsequently, the 96-well plate was removed, the original culture medium was removed, and a phosphate buffered saline (PBS) wash was performed. The experiment was divided into a background control group, a solvent control group, and a drug treatment group. The experimental settings included a background control group (cell-free low serum culture medium with a serum concentration of 2%), a solvent control group (DMSO treatment), FICZ (10 -6 mol / L) treatment group, ginsenoside Rc(10 -6 mol / L) treatment group and FICZ (10 -6 mol / L) and ginsenoside Rc(10 -6 After drug addition, the 96-well plates were incubated in 37°C and 38.5°C incubators for 24 hours. At the end of the incubation period, 10 μL of CCK8 reagent was added to each well according to the kit instructions. The plates were then placed in the incubator and incubated for 1 hour. The absorbance was then read using a microplate reader.

[0044] Effects of ginsenoside Rc and FICZ on the activity of HaCaT cells treated with elevated temperature Figure 1 shown.

[0045] Figure 1 The results showed that each treatment group had no significant effect on the viability of HaCaT cells within 24 hours, indicating that under this experimental condition, ginsenoside Rc and FICZ had no toxic effects on cells and were suitable for subsequent studies.

[0046] Example 2. Ginsenoside Rc reduces the gene expression levels of IL-6 and IL-8 in HaCaT cells under elevated temperature conditions

[0047] HaCaT cells were grown at 2×10 5 Each well of the 6-well culture plate was evenly distributed with 2 mL of culture medium. After culturing in the culture plate for 24 hours, the original culture medium was removed and replaced with a medium containing DMSO, FICZ (10 -6 mol / L), ginsenoside Rc(10 -6 mol / L)、FICZ(10 -6 mol / L) and ginsenoside Rc(10 -6 mol / L) mixed culture medium. A total of 4 treatment groups were set up, namely DMSO, FICZ (10 -6 mol / L), ginsenoside Rc(10 -6 mol / L)、FICZ(10 -6 mol / L) and ginsenoside Rc(10 -6 mol / L) were mixed with the culture medium of the treatment groups. After further culture in a 37°C or 38.5°C incubator for 24 hours, the culture medium was aspirated, and the cells in the six-well plate were gently washed with 1 mL of PBS. Then, 1 mL of lysis buffer was added to each well, along with 20 μL of β-mercaptoethanol. The mixed lysis buffer (600 μL per well) was used to lyse the cells, and the cell lysate was collected in an EP tube.

[0048] Using Gene Total RNA was extracted from cultured cells using an RNA purification kit (Thermo Fisher Scientific, Waltham, MA, USA). The quality and concentration of RNA were determined using a NanoDrop 2000 / 2000c spectrophotometer (Thermo Fisher Scientific). RNA was reverse transcribed into cDNA using a reverse transcription-assisted first-strand cDNA synthesis kit (Thermo). TM 6 Using mRNA in FlexReal-TimePCR TMTarget mRNA was amplified in 20 μL reactions using the 6Flex Real-Time PCR System (ThermoFisher Scientific). qRT-PCR Master Mix (Promega, Madison, WI, USA) was used. Relative quantities were calculated by normalizing the mean threshold cycle (Ct) values ​​to a reference gene (GAPDH) to generate the ΔCt of the target gene, and the relative fold change was calculated by subtracting the control Ct value to obtain 2-ΔΔCt.

[0049] GAPDH (F: 5'ATT GTT GCC ATC AAT GAC CC 3' (SEQ ID NO. 1); R: 5'AGT AGA GGCAGG GAT GAT GT 3' (SEQ ID NO. 2)); IL-6 (F: 5'CCT GAC CCAACC ACAAAT GC 3' (SEQ ID NO. 3); R: 5'ATC TGA GGT GCC CAT GCT AC 3' (SEQ ID NO.4)); IL-8 (F: 5'ACT GAG AGTGAT TGA TGG AC 3' (SEQ ID NO. 5); R: 5'AAC CCT CTG CAC CCA GTT TTC 3' (SEQ ID NO. 6)).

[0050] Figure 2 The results of the experiments showed that ginsenoside Rc significantly reduced the gene expression levels of IL-6 and IL-8 under experimental conditions of 38.5°C. These data emphasize the effectiveness of ginsenoside Rc as a potential anti-inflammatory agent.

[0051] Example 3. Ginsenoside Rc reduces the protein secretion levels of IL-6 and IL-8 in HaCaT cells under elevated temperature conditions

[0052] HaCaT cells were grown at 2×10 5 Each well of the 6-well culture plate was evenly distributed with 2 mL of culture medium. After culturing in the culture plate for 24 hours, the original culture medium was removed and replaced with a medium containing DMSO, FICZ (10 -6 mol / L), ginsenoside Rc(10 -6 mol / L)、FICZ(10 -6 mol / L) and ginsenoside Rc(10 -6 mol / L) mixed culture medium. There were 4 treatment groups, namely DMSO, FICZ (10 -6 mol / L), ginsenoside Rc(10 -6 mol / L)、FICZ(10-6 mol / L) and ginsenoside Rc(10 -6 mol / L) were mixed with the culture medium of the treatment group. After incubation at 37°C and 38.5°C for 24 hours, the supernatant was collected. The IL-6 and IL-8 levels in the supernatant were determined according to the instructions of the ELISA kit.

[0053] Figure 3 The experimental results showed that at 38.5°C, ginsenoside Rc significantly reduced the secretion of IL-8 protein in heat-induced HaCaT cells in both the DMSO and FICZ treatment groups, and also showed a downward trend in IL-6 protein expression. These findings suggest that ginsenoside Rc may have a potential therapeutic effect in responding to inflammation induced by high temperature environments, indicating its application prospects in the anti-inflammatory field.

Claims

1. Application of ginsenoside Rc in the preparation of products for resisting skin high temperature and light damage; The structural formula of ginsenoside Rc is shown in (Formula I):

2. The use according to claim 1, characterized in that: The resistance to skin high temperature and light damage is reflected in the following aspects: (1) resistance to inflammation induced by high temperature; (2) resistance to inflammation induced by high temperature and ultraviolet light product FICZ.

3. The use according to claim 1 or 2, characterized in that: The inflammation is caused by high temperature inducing skin cells to secrete inflammatory factors IL-6 and / or IL-8.

4. The use according to claim 1 or 2, characterized in that: The inflammation is caused by high temperature and ultraviolet light product FICZ inducing skin cells to secrete inflammatory factors IL-6 and / or IL-8.

5. The use according to claim 2, characterized in that: The method of resisting inflammation induced by high temperature or the method of resisting inflammation induced by high temperature and ultraviolet light product FICZ is to inhibit the gene expression level and / or protein expression level of inflammatory factors IL-6 and IL-8 by ginsenoside Rc.

6. The use according to any one of claims 1 to 5, characterized in that: The product is a medicine and / or a cosmetic.

7. A product for resisting skin damage caused by high temperature and light, wherein the active ingredient thereof includes ginsenoside Rc.

8. The product according to claim 7, characterized in that: The product is a medicine and / or a cosmetic.

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