The Effects Of5-Androstenedione、Luteinizing Hormone And Insulin On The Anti-Mullerian Hormone Secretion Of Human Granulosa Cells

Objective:To investigate the effects of5-Androstenedione (A)、Luteinizing Hormone (LH) and Insulin (INS) on the anti-mullerian hormone (AMH) secretion of human granulosa cells in vitro.Methods:Ovarian luteinizing granulosa cells from12womeu with normal cycles and infertility caused by structural and male disorders, who were undergoing oocyte retrieval as part of the in—vitro fertilization/embryo transfer program. Human luteinized granulose cells were cultured according to literature. Granulose cells were inoculated into24-well plates at a density of1.0x105viable cells/lml/well with DMEM medium containing10%FBS. The medium was replaced by serum—free medium after24hour. Ovarian luteinizing granulosa cells were treated with different concentrations ofA(I A1×10-4mol/L,ⅡA1×10-5mol/L,Ⅲ A1×10-6mol/L). LH(Ⅰ LH10ng/ml, Ⅱ LH5ng/ml, Ⅲ LH lng/ml)、INS (Ⅰ INS800Uiu/ml,Ⅱ INS400Uiu/ml,Ⅲ INS200Uiu/ml)、A+LH (Ⅱ A+ⅠLH,Ⅱ ⅡA+Ⅱ LH,Ⅲ Ⅲ A+Ⅲ LH)、A+INS (Ⅰ Ⅰ A+Ⅰ INS, Ⅱ ⅡA+Ⅱ INS, Ⅲ Ⅲ A+Ⅲ INS) and LH+INS((Ⅰ Ⅰ LH+Ⅰ INS, Ⅱ ⅡLH+Ⅱ INS, Ⅲ Ⅲ LH+Ⅲ INS) for48hour, AMH contents in the culture media were detected by ELISA kit.Results:As compared with those in blank control group, AMH contents in the culture media of A、A+LH and A+INS groups were significantly difference. AMH level was relatively higher with concentrations of A. A+LH and A+INS in the culture media increasing; In addition, no significant difference of AMH contents in LH、INS and LH+INS groups was found.Conclusion:AMH secretion of granulosa cells obtained was influened by androgen,and was not influened by LH and INS, AMH production arised with the uprise of androgen. LH may have coordinated in AMH secretion of human granulosa cells with androgen.