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目的探讨养精胶囊改善生精功能进而治疗男性不育症可能的作用机制。方法在GC-1细胞中加入养精胶囊提取液,分为空白组和低、中、高剂量组,在作用24、48、72 h后采用MTT法检测各组的吸光值;作用48 h后,采用流式细胞仪检测细胞周期及细胞凋亡;用实时荧光定量PCR法和Western blot法检测凋亡相关基因caspase-3的表达。结果在作用24、48、72 h后,养精胶囊提取液低、中、高剂量组细胞增殖明显加快,其吸光度与同期空白组相比有统计学意义(P<0.01);养精胶囊提取液低、中、高剂量组作用GC-1细胞48 h后,S期细胞比例逐渐增加,与空白组相比有统计学意义(P<0.05);细胞凋亡率明显降低,与空白组相比有统计学意义(P<0.01);caspase-3mRNA表达呈下降趋势,中、高剂量组与空白组相比有统计学意义(P<0.01);caspase-3蛋白表达呈下降趋势,高剂量组与空白组相比有统计学意义(P<0.01)。结论养精胶囊具有促进GC-1细胞增殖、抑制细胞的凋亡,降低caspase-3的表达水平的作用,且呈剂量依赖性。
Objective To investigate the possible mechanism of Yangjing Capsule in improving the function of spermatogenesis and further treating male infertility. Methods The extract of Yangjing Capsule was added into GC-1 cells and divided into blank group and low, medium and high dose groups. After 24, 48 and 72 hours, the absorbance of each group was detected by MTT assay. After 48 hours The cell cycle and apoptosis were detected by flow cytometry. The expression of caspase-3 was detected by real-time fluorescence quantitative PCR and Western blot. Results After 24h, 48h and 72h, the cell proliferation of Yangzhong capsule extract in low, medium and high dose groups was significantly accelerated, and the absorbance was significantly higher than that in the blank control group (P <0.01) Compared with the blank group, the percentage of S phase cells increased gradually in GC-1 cells treated with low, medium and high dose of GC-1 for 48 h (P <0.05) (P <0.01). The expression of caspase-3 mRNA showed a decreasing trend. The expression of caspase-3 protein in middle-dose and high-dose groups was significantly lower than that in the blank group (P <0.01) Compared with the blank group, there was statistical significance (P <0.01). Conclusion Yangjing Capsule can promote the proliferation of GC-1 cells, inhibit the apoptosis and decrease the expression of caspase-3 in a dose-dependent manner.