Application of ginsenoside Rg2 in preparation of medicine for resisting renal cell injury caused by cisplatin chemotherapy

Abstract

The invention discloses an application of ginsenoside Rg2 in preparation of a drug for resisting cisplatin chemotherapy-induced renal cell injury, and discovers that the ginsenoside Rg2 has a unique effect on improvement of cisplatin-induced acute renal cell injury, and can relieve kidney cell killing caused by cisplatin chemotherapy without affecting cancer treatment. The ginsenoside Rg2 can be used as an auxiliary drug for cancer cis-platinum chemotherapy, and a new medication strategy is provided for relieving toxic and side effects of chemotherapy of cancer patients.

Description

Technical Field

[0001] The present invention relates to a new application of ginsenoside Rg2, in particular to an application of ginsenoside Rg2 in preparing a drug for resisting cisplatin chemotherapy-induced renal cell damage. Background Art

[0002] Cisplatin (cDDP) is the full name of cis-1,2-dichlorodiammineplatin. It is a non-specific drug for the cell cycle and has cytotoxicity. It can inhibit the DNA replication process of cancer cells and damage their cell membrane structure. It has a strong broad-spectrum anti-cancer effect and is currently the most widely used anti-cancer drug in medicine. However, cisplatin accumulates in large quantities in the body and has side effects such as vomiting, ototoxicity, neurotoxicity, nephrotoxicity, and allergic reactions. Among them, nephrotoxicity is the most significant. According to statistics, the incidence of renal damage caused by clinical cisplatin chemotherapy is about 30%. The mechanism of cisplatin-induced renal damage is not very clear, but it can be determined that the process is complex, which may involve cytotoxicity, oxidative damage, inflammatory response, apoptosis, mitochondrial dysfunction and autophagy. At present, no drug has been found that can effectively treat cisplatin-induced renal toxicity.

[0003] Ginseng (Panax ginseng CA Meyer.) is the dried root and rhizome of the genus Panax in the Araliaceae family. It is a precious Chinese herbal medicine. Its main active ingredients are ginsenosides, ginseng polysaccharides, ginsenoside A, etc. It has multiple pharmacological effects such as anti-tumor, anti-aging and improving the body's immunity. Existing technology shows that the subtle differences in the chemical structures of different ginsenosides have a significant impact on their biological functions. Even for ginsenosides with the same molecular formula, the difference in optical rotation alone can lead to huge differences in their chemical activities.

[0004] Ginsenoside Rg3 can significantly inhibit the growth of tumors in liver cancer mouse models. When used in combination with cisplatin, it can enhance the chemotherapy effect and inhibit the toxicity of cisplatin to a certain extent. At the same time, ginsenoside Rg3 can increase the sensitivity of gastric cancer cell lines to cisplatin by downregulating the PD-L1 gene, thereby inhibiting the proliferation of gastric cancer cells.

[0005] The combination of ginsenoside Rh2 and cisplatin in a prostate cancer mouse model not only reduces side effects but also induces cancer cell apoptosis by significantly enhancing caspase-3 mRNA expression. In vitro low-dose ginsenoside Rh2 may increase the intracellular cisplatin concentration of human esophageal cancer cells by reducing intracellular p53 expression, thereby enhancing the killing effect of cisplatin.

[0006] Ginsenoside Rg5 can synergistically inhibit the growth of esophageal cancer cells and promote cell apoptosis with cisplatin, and its mechanism may be related to the downregulation of Bcl-2 and p-AKT protein expression.

[0007] Ginsenoside Rg2 has a protective effect on the nervous system and cardiovascular and cerebrovascular systems. The invention patent application with Chinese patent application number 201110126282.6 discloses an application of ginsenoside Rg2 in the preparation of anticoagulant drugs; the invention patent with Chinese patent application number 201810970997.1 discloses an application of ginsenoside Rg2 in the preparation of products for preventing and treating arrhythmias.

[0008] To date, although the inhibitory effect of ginsenoside Rg2 on cancer cell activity has been reported, there are no reports on the combined use of ginsenoside Rg2 and cisplatin in the treatment of cancer, nor are there any reports on ginsenoside Rg2 in resisting cisplatin-induced renal damage. Summary of the invention

[0009] The present invention is based on the discovery that ginsenoside Rg2 has the effect of improving cisplatin-induced acute renal cell injury, thereby inventing an application of ginsenoside Rg2 in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell injury.

[0010] The technical solution of the present invention is: an application of ginsenoside Rg2 in the preparation of a drug for resisting cisplatin-induced renal cell damage.

[0011] Preferably, ginsenoside Rg2 is used in combination with cisplatin in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell damage.

[0012] Preferably, the concentration ratio of ginsenoside Rg2 to cisplatin is 20-80 μg / ml:10-20 μM.

[0013] Preferably, the concentration ratio of ginsenoside Rg2 to cisplatin is 20 μg / ml: 10-20 μM.

[0014] Preferably, the pharmaceutical purity of the ginsenoside Rg2 is at least 98%.

[0015] The present invention discovered that ginsenoside Rg2 has a unique effect in improving cisplatin-induced acute renal cell injury. It can alleviate the kidney cell killing caused by cisplatin chemotherapy without affecting the treatment of cancer, so that ginsenoside Rg2 can be used as an auxiliary drug for cisplatin chemotherapy of cancer, providing a new medication strategy for reducing the toxic side effects of chemotherapy in cancer patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Figure 5 is a graph showing the half inhibitory concentration (IC50) of kidney HEK293 cells after 24 hours of cisplatin treatment.

[0017] Figure 2 This is a diagram showing the half inhibitory concentration (IC50) of kidney HK-2 cells after 24 hours of cisplatin treatment.

[0018] Figure 3 This is a diagram showing the effect of ginsenoside Rg2 combined with cisplatin on renal HEK293 cell damage caused by anti-cisplatin chemotherapy.

[0019] Figure 4 This is a diagram showing the effect of ginsenoside Rg2 combined with cisplatin on kidney HK-2 cell damage caused by anti-cisplatin chemotherapy. DETAILED DESCRIPTION

[0020] Experiment on the effect of ginsenoside Rg2 on renal cell damage caused by cisplatin chemotherapy: 1. Sources of Materials and Reagents Kidney HEK293 cells (abbreviated as HEK293) and kidney HK-2 cells (abbreviated as HK-2) were stored in our laboratory and purchased from the American type culture collection (ATCC) cell bank.

[0021] DMEM and MEM culture media were purchased from Hyclone, USA, fetal bovine serum was purchased from Gibco, USA, cisplatin (cDDP) was purchased from Sigma, USA, trypsin was purchased from Hyclone, USA, DMSO was purchased from Sigma, USA, CCK8 kit was purchased from Beijing Weibai’ao Biotechnology Co., Ltd., and ginsenoside Rg2 was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with a purity greater than 98%.

[0022] 2. Experimental drug preparation 1) Use DMSO as solvent to fully dissolve ginsenoside Rg2 and prepare a 20 mg / mL stock solution; 2) After the mother solution is prepared, use culture medium to dilute the mother solution to the required 20μg / ml, 40μg / ml, and 80μg / ml.

[0023] 3. Experimental Methods 3.1 Cell culture HEK293 cells were cultured in DMEM medium containing 10% fetal bovine serum, 1% penicillin, 1% streptomycin and 1% amphotericin B. HK-2 cells were cultured in MEM (containing NEAA) + 10% FBS + 1% P / S. The cells were placed in a cell culture incubator at 37°C and 5% CO2. When the cells reached 70-80% confluence, they were digested and passaged with trypsin. Cells in the logarithmic growth phase and in good condition were used for subsequent experiments.

[0024] 3.2 Cell viability inhibition experiment with cisplatin alone HEK293 cells and HK-2 cells in the logarithmic growth phase were selected, digested with 0.25% trypsin, centrifuged at 1000 rpm for 5 min, the supernatant was discarded, the above culture medium was added, the cells were evenly blown, the cell concentration was adjusted to 1000 cells / well, and the cells were evenly inoculated in a 96-well plate. After culturing in an incubator for 24 hours, the complete culture medium was discarded and the drug-added culture medium was replaced. The control group was complete culture medium and cells, the experimental group was complete culture medium containing different concentrations of cisplatin, and the blank group was the same volume of complete culture medium. After culturing for 24 hours, 10 μL CCK8 was added to each well and incubated in the dark for 1 hour, and the absorbance (OD) value of the well plate at a wavelength of 450 nm was measured by an enzyme reader. Cell viability inhibition rate (Inhibition Rate) = 1-[A(drug added)-A(blank)] / [A(control)-A(blank)]. Half maximal inhibitory concentration (IC50) of cell viability was calculated using GraphPad Prism 5.02 software (GraphPad Software, San Diego CA, USA).

[0025] 3.3 Cell viability inhibition experiment with cisplatin and ginsenoside Rg2 HEK293 cells and HK-2 cells in the logarithmic growth phase were selected, digested with 0.25% trypsin, centrifuged at 1000 rpm for 5 min, the supernatant was discarded, the above culture medium was added, the cells were evenly pipetted, the cell concentration was adjusted to 1000 cells / well, and the cells were evenly inoculated in a 96-well plate. After culturing in an incubator for 24 hours, the complete culture medium was discarded and the drug-added culture medium was replaced. The control group was complete culture medium and cells, the experimental group was complete culture medium containing cisplatin (20 μM for HEK293 cells and 10 μM for HK2 cells) reaching or exceeding IC50 and different concentrations of ginsenoside Rg2 (0 μg / ml, 20 μg / ml, 40 μg / ml, 80 μg / ml), and the blank group was the same volume of complete culture medium. After culturing for 24 hours, 10 μL CCK8 was added to each well and incubated in the dark for 1 hour, and the absorbance (OD) value of the well plate at a wavelength of 450 nm was measured using an enzyme reader. Cell viability = [A(drug added)-A(blank)] / [A(control)-A(blank)].

[0026] 4. Experimental Results 4.1 Schematic diagram of the half inhibitory concentration (IC50) of kidney HEK293 cells treated with cisplatin for 24 hours Figure 1 As shown. Figure 1It can be seen that the half inhibitory concentration (IC50) of kidney HEK293 cells treated with cisplatin for 24 hours was 20.4 μM.

[0027] 4.2 Schematic diagram of the half inhibitory concentration (IC50) of kidney HK2 cells after 24 hours of cisplatin treatment Figure 2 As shown. Figure 2 It can be seen that the half inhibitory concentration (IC50) of kidney HK2 cells treated with cisplatin for 24 hours is 7.5 μM.

[0028] 4.3 Effect of ginsenoside Rg2 combined with cisplatin on renal HEK293 cell damage caused by anti-cisplatin chemotherapy Figure 3 As shown. Figure 3 It can be seen that the addition of different concentrations of ginsenoside Rg2 (20 μg / ml, 40 μg / ml, 80 μg / ml) effectively improved cisplatin-induced acute renal HEK293 cell injury, and the optimal concentration of ginsenoside Rg2 used was 20 μg / ml.

[0029] 4.4 Effect of ginsenoside Rg2 combined with cisplatin on HK2 cell damage in kidney caused by anti-cisplatin chemotherapy Figure 4 As shown. Figure 4 It can be seen that the addition of different concentrations of ginsenoside Rg2 (20 μg / ml, 40 μg / ml, 80 μg / ml) effectively improved cisplatin-induced acute renal HK2 cell injury, and the optimal concentration of ginsenoside Rg2 used was 20 μg / ml.

[0030] In summary, the experimental results of the present invention show that: The combination of ginsenoside Rg2 and cisplatin significantly improved the inhibitory effect of cisplatin on the vitality of different types of kidney cells, suggesting that ginsenoside Rg2 has the effect of alleviating the nephrotoxicity caused by cisplatin chemotherapy and can be used as an auxiliary drug for cisplatin chemotherapy for cancer.

Claims

1. An application of ginsenoside Rg2 in the preparation of a drug to resist cisplatin chemotherapy-induced renal cell damage.

2. The use of ginsenoside Rg2 according to claim 1 in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell damage, characterized in that Application of ginsenoside Rg2 combined with cisplatin in the preparation of drugs against cisplatin chemotherapy-induced renal cell damage.

3. The use of ginsenoside Rg2 according to claim 2 in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell damage, characterized in that The concentration ratio of ginsenoside Rg2 and cisplatin is 20-80 μg / ml: 10-20 μM.

4. The use of ginsenoside Rg2 according to claim 3 in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell damage, characterized in that The concentration ratio of ginsenoside Rg2 and cisplatin is 20 μg / ml: 10-20 μM.

5. The use of ginsenoside Rg2 according to claim 1, 2, 3 or 4 in the preparation of a drug for resisting cisplatin chemotherapy-induced renal cell damage, characterized in that The pharmaceutical purity of the ginsenoside Rg2 is at least 98%.

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