目的:观察体外培养的大鼠窦前期卵泡颗粒细胞中端粒酶表达的调控因素。方法:体外培养大鼠窦前期卵泡颗粒细胞,加入不同处理因素振荡培养,用端粒酶重复扩增酶联免疫吸附分析法(TRAP-ELISA法)分析端粒酶活性的改变。结果:在卵泡颗粒细胞中表达有端粒酶活性,且在人绒毛膜促性腺激素(hcG)、促卵泡生成素(FSH)、二丁酰环磷腺苷(dbcAMP)及维拉帕米(verapamil)作用下活性明显升高,而在反义c-myb作用下活性明显降低。同时用放射免疫法测定培养液中雌孕激素含量发现,在Verapamil及FSH作用下雌孕激素分泌量明显升高,在db-cAMP及hcG作用下分泌量无明显改变,而在反义c-myb作用下分泌量明显降低,在不同作用因素下的端粒酶活性与它相对应的E2分泌量成正相关r=0.953,P〈0.01。用四甲基偶氮唑盐(MTT)法测定,反义hTERT能明显抑制颗粒细胞的增殖。结论:窦前期卵巢的颗粒细胞中表达有端粒酶活性,强弱可进行调控。
Aim: To study the expression and control of telomerase in rat preantral ovary. Methods: Added different factors to the preantral ovarian granulosa cells, then using TRAP-ELISA to analyze the expression of telomerase. Results: Telomerase activation was detected in granulosa cells. Telomerase activation could be significantly enhanced by heG, FSH, verapamil and dbcAMP, and it was signifi- cantly reduced by antisense-c-myb ODN treatment. We also used radi y to determine proteronse and estrogen in these cell culture mediums. It was found that these two hormones' secretion were raised under verapamil and FSH; no change under heG and dbcAMllted with telomerase activity. In MTT assay, the antisense hTERT ODN could significantly inhibited the proliferation of o- varian granulosa cells.Conclusion: It is suggested that telomerase activation is present in ovary antral granulosa cell. Anti-c-myb might inhibit telomerase activation, while FSH, hcG, verapamil, dbcAMP might enhance the telomerase activation.