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目的探讨养精胶囊促进睾丸间质(Leydig)细胞合成睾酮(T)功能的具体作用机制。方法在Leydig细胞(MLTC-1细胞系)中加入不同剂量养精胶囊提取液24 h后,用化学发光法检测细胞上清T浓度;用免疫荧光显微镜分析类固醇快速调节蛋白(StAR)的表达情况;用RT-PCR及Western blot法测StAR mRNA及蛋白质的表达;用双荧光素酶报告基因实验检测StAR启动子的活性。结果低、中、高剂量养精胶囊提取液作用24 h后,均可以促进MLTC-1细胞合成睾酮的功能;养精胶囊提取液可以促进MLTC-1细胞StAR免疫荧光的表达;养精胶囊提取液可以促进MLTC-1细胞StAR mRNA和蛋白质的表达;养精胶囊提取液可以提高MLTC-1细胞中StAR启动子的活性。结论养精胶囊能通过调控Leydig细胞中StAR启动子的活性,促进StAR mRNA和蛋白的表达,进而提高睾酮合成的功能。
Objective To investigate the mechanism of Yangjing Capsule in promoting testosterone (T) function in Leydig cells. Methods Twenty-four hours after adding different doses of Yangjing Capsule extract to Leydig cells (MLTC-1 cell line), the concentration of T in the supernatant of the cells was detected by chemiluminescence method. The expression of StAR was analyzed by immunofluorescence microscopy The expression of StAR mRNA and protein was detected by RT-PCR and Western blot, and the activity of StAR promoter was detected by dual luciferase reporter assay. Results After 24 h treatment, MLTC-1 cells could promote the function of testosterone synthesis in MLTC-1 cells. SFE capsule could promote the expression of StAR immunofluorescence in MLTC-1 cells. Liquid can promote the expression of StAR mRNA and protein in MLTC-1 cells. The capsule extract can increase the activity of StAR promoter in MLTC-1 cells. Conclusion Yangjing Capsule can improve the function of testosterone synthesis by regulating the activity of StAR promoter in Leydig cells and promoting the expression of StAR mRNA and protein.