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In this study,we investigated the effects of the Yangjing capsule extract,which is used to treat male sexual dysfunction and infertility,on steroidogenesis and apoptosis in testicular Leydig cells.Mouse Leydig tumor cells (MLTC-1) were cultured and treated with Yangjing capsule extract (0.01-1 mg/mL) for 24 h.The testosterone levels in the medium were evaluated by radioimmunoassay.Expression of steroidogenic key enzymes (StAR,CYP11A1,HSD3B) was examined using real-time RT-PCR and immunoblotting.Subsequently,MLTC-1 were treated with Yangjing capsule extract (0.01-1 mg/mL) for 48 h in a serum-free medium.Cell viability was measured by MTT assays.The cell cycle and apoptosis were analyzed using flow cytometry.The expression of activated caspase-3 was analyzed using RT-PCR and a colorimetric protease assay.Yangjing capsule extract showed higher testosterone production and the expression of StAR,CYP11A1,and HSD3B mRNAs and proteins than those in control group in MLTC-1.In addition,H89,a PKA inhibitor,significantly inhibited Yangjing-stimulated testosterone production.In another experiment,Yangjing capsule extract improved the cell viability of MLTC-1 and decreased the number of cells in the G0/G1 phase while increasing the number of cells in the S-phase in a dose-dependent manner.Further investigation showed that Yangjing capsule extract prevented cell apoptosis by inhibiting the expression of caspase-3.These results showed that Yangjing capsule extract can improve steroidogenesis and inhibit apoptosis in MLTC-1.