Effects And Mechanisms Of Yangjing Capsule Extract On Testosterone Production In Leydig Cells

Objective:To explore the effects and mechanisms of Yangjing capsule extract on testosterone production in testicular Leydig cells.Methods:Mouse Leydig tumor cells (MLTC-1) were cultured and treated by Yangjing capsule extract (0.01～1mg/mL) for24h. Testosterone levels in medium were evaluated by radioimmunoassay. Expressions of steroidogenic key enzymes (StAR, CYP11A1, HSD3B2) were examined by Real-time RT-PCR and immunoblotting. Then MLTC-1cells were treated by Yangjing capsule extract (0.01～1mg/mL) for48h in serum free medium. Cell viability was measured by MTT assays. Cell cycle and apoptosis were analyzed by flow cytometry. Expression of the activated caspase-3was analyzed by real-time RT-PCR and colorimetric protease assay.Results:Yangjing capsule extract increased the testosterone production. H89, PKA inhibitor, significantly inhibited Yangjing-stimulated testosterone production. Yangjing capsule extract improved the expression of StAR, CYP11A1, HSD3B2mRNAs and proteins in MLTC-1cells. In another side, Yangjing capsule extract improved the cell viability and decreased cells in G0/G1phase while increased cells in S-phase of MLTC-1cells in a dose-dependent manner. Further study showed Yangjing capsule extract prevented cell apoptosis via inhibiting the expression of caspase-3.Conclusion:Yangjing capsule extract can improve the steroidogenesis and inhibit apoptosis to increase the testosterone production in MLTC-1cells.