激活素A/抑制素A在LH峰效应的表皮生长因子信号通路中的作用机制  

Mechanism of Activin A/Inhibin A in Epidermal Growth Factor Signaling Pathway of LH Surge Effect

作　　者：陈颖[1] 李鹏[2] 袁平[2] 肖露[1] 锦凤 钱卫平[1] 

机构地区：[1]北京大学深圳医院生殖医学科,深圳518036 [2]香港中文大学深圳研究院,深圳518057

出　　处：《生殖与避孕》2014年第11期881-886,共6页Reproduction and Contraception

基　　金：国家自然科学基金青年基金项目;项目号:81100465

摘　　要：目的:探讨激活素A(ACTA)/抑制素A(INHA)在LH峰效应的表皮生长因子(EGF)信号通路中的作用机制。方法:1经孕马血清促性腺激素(PMSG)预处理后48 h,取小鼠卵丘-卵母细胞复合物(COCs),随机置入含0μg/L、5μg/L、10μg/L、20μg/L EGF的体外成熟体系中进行体外成熟(IVM)培养,2 h后收集卵丘颗粒细胞,进行Real-time PCR(RT-PCR)检测各组颗粒细胞中ACTβA亚基,ACTRⅡ、INHα亚基的表达水平。2上述体外培养体系中加入表皮生长因子受体(EGFR)的强抑制剂AG1478后,检测各组颗粒细胞中ACTβA亚基、ACTRⅡ、INHα亚基的表达水平。3 COCs分别置入含10μg/L EGF的体外成熟体系中1 h、2 h、3 h、4 h,进行IVM培养,收集卵丘颗粒细胞,提取RNA,用RT-PCR检测各组颗粒细胞中ACTβA亚基、ACTRⅡ、INHα亚基的表达水平。结果:1 EGF可刺激小鼠卵丘颗粒细胞ACTβA亚基的表达,抑制INHα亚基的表达,而对ACTRⅡ型受体m RNA的表达无明显影响。2 EGF对ACT/INHβA亚基的促进作用以及对INHα亚基的抑制作用具有时效性,在作用后2 h达到最强,随后逐渐减弱。3 EGF对ACT/INHβA亚基的促进作用以及对INHα亚基的抑制作用具有剂量依赖性,10μg/L时的作用效力最强。结论:说明ACTA/INHA是卵巢内部EGF网络的下游信号分子,而EGF对两者具有不同的调控方向,并呈现显著的时间依赖性和剂量依赖性。Objective： To investigate the mechanism of activin A/inhibin A （ACTA/INHA） in epidermal growth factor （EGF） signaling pathway of LH surge effect. Methods： 1） Cumulus-oocyte complex （COCs） fromPMSG-prirned 3-4 weeks old mice were isolated at GV stage. COCs were in vitro maturation （IVM） cultured in the control and 5 μg/L, 10 μg/L, 20 μg/L EGF treatment media. After 2 h, granular cells were collected. The mRNA of ACTβA subunit, ACTR Ⅱ, INHα subunit were detected by real-time PCR. 2） EGFR inhibitor AG1478 was added into the IVM media. Expression of mRNA ofACTβA subunit, ACTR Ⅱ, INHα subunit were detected. 3） COCs were IVM cultured in 10 μg/L EGF treatment media, 1 h, 2 h, 3 h, 4 h later, granular cells were collected. The mRNA ofACβA subunit, ACTR Ⅱ, INHot subunit were detected by real-time PCR. Results： 1） EGF can significantly increase the expressin ofACTβA subunit, inhibit the expression oflNHα subunit. There was no significant effect on the expression ofACTR Ⅱ mRNA. 2） The stimulation effect of EGF on ACTβA subunit and supression effect on INHα subunit were time-dependent and dose-dependent. Two hours after treatment at the dose of 10 μg/L were the optimal time and dose. Conclusion： ATCA/INHA are the downstream of EGF signaling pathway. EGF exerts opposite regulation on ACTA and 1NHA. It indicates that a certain ratio of ACTA/INHA may be appropriate for IVM culture of oocyte.

关 键 词：激活素A(ACTA) 抑制素A(INHA) 表皮生长因子(EGF) LH峰 

分 类 号：R714.8[医药卫生—妇产科学]