Preparation method of 20(r)-ginsenoside rg3 and its application in the preparation of heat stress-induced spermatogenic damage medicine
A technology of ginsenoside and heat stress, which is applied in the field of medicine for the development of natural drug activity, to achieve the effect of remarkable efficacy, less dosage, and low-cost acquisition
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experiment example 1
[0027] Experimental Example 1: Preparation and isolation of R-Rg3 using ginsenoside Rd as a substrate
[0028] 1.1 Single factor investigation experiment
[0029] 1.1.2 The effect of acid concentration on the reaction
[0030] Accurately weigh 20 mg of ginsenoside Rd powder and dissolve in 1 mL of formic acid solutions with concentrations of 0.05%, 0.1%, and 0.2%, respectively, and react at 100° C. for 3 h. After the reaction, the sample was centrifuged to obtain a precipitate and the supernatant was concentrated. Part of the precipitate was removed and the methanol was settled in a 10 mL volumetric flask for HPLC analysis. The content of rare ginsenoside Rg3 in the precipitate was calculated according to the peak area. The results are shown in Table 1. It can be seen from the results that the content of Rg3 is low at 0.05%, and there is incomplete hydrolysis of Rd, while the generation of rare ginsenoside Rg3 is at a maximum at 0.1%, and the ratio of by-product Rk1 / Rg5 is lo...
experiment example 2
[0044] Experimental example 2: Protective effect of R and S configuration ginsenoside Rg3 on mouse Leydig cells TM3
[0045] The experimental method refers to the literature (Zhao Jian, et al. Establishment of oxidative stress model of TM3 cells induced by hydrogen peroxide, China Public Health, 2015, 31, 312-314). The specific experimental scheme is omitted.
[0046] As shown in Table 4, the experimental results show that the two configurations of Rg3 were applied to TM3 cells at three doses for 24 hours, and the dosage of hydrogen peroxide was 600 μM. configuration, especially the effect of 6μM in the high-dose group was more obvious.
[0047] According to literature reports and test results of the present invention (Hee-Won Park, et al., J Ginseng Res2013,37,457-467; Jin Yan, et al., Plant Foods Hum Nutr, 2015,70:141-145; Lee, et al. ., J.Ginseng Res.2012,36,102-106), the ratio of S configuration to R configuration Rg3 in red ginseng is about 2:1~3:1, as attached figure ...
experiment example 3
[0054] Experimental Example 3: The protective effect of 20(R)-ginsenoside Rg3 on the damage of germ cells induced by heat stress in the scrotum
[0055] 3.1 Materials and methods
[0056] 3.1.1 Materials, reagents and instruments
[0057] 20(R)-ginsenoside Rg3 (hereinafter referred to as Rg3), purity > 98.5% (HPLC); sodium pentobarbital (C 11 h 17 N 2 NaO 3 ), purity>99%.
[0058]Malondialdehyde (MDA), reduced glutathione (GSH) and superoxide dismutase (SOD) kits were purchased from Nanjing Jiancheng Bioengineering Institute; testosterone (Testosterone) kits were purchased from American R&D Corporation . BP211D electronic balance, Germany Sartorius company; DK-98-1 type electric heating constant temperature water bath, Tianjin Test Instrument Co., Ltd.; HC-2517 high-speed centrifuge, Anhui Zhongke Zhongjia Company; Epoch ultra-micro microplate spectrophotometer , American Boten Instrument Co., Ltd.; FSH-2A adjustable high-speed homogenizer, Jintan Huacheng Hailong Exper...
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