摘要
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.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2008年第1期86-89,共4页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(30260035)
关键词
端粒
端粒酶
窦前期
颗粒细胞
表达
调控
telomere
telomerase
preantral granulosa cell
distribution and control
