IP3R在LH-EGFR诱导小鼠卵母细胞第一次减数分裂恢复中的作用  

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

作　　者：郝肖琼 李延康 付旭阳 贾方毅 HAO Xiao-qiong;LI Yan-kang;FU Xu-yang;JIA Fang-yi(Department of Physiology,Baotou Medical College;Inner Mongolia Baogang Hospital,Baotou 014040,China)

机构地区：[1]包头医学院生理学教研室 [2]内蒙古包钢医院,内蒙古包头014040

出　　处：《中国病理生理杂志》2020年第3期532-538,共7页Chinese Journal of Pathophysiology

基　　金：内蒙古自治区自然科学基金资助项目(No.2018BS03022);内蒙古自治区高等学校科学研究项目(No.NJZY18188);内蒙古自治区高等学校“青年科技英才支持计划”项目(No.NJYT-19-B23);包头市医药卫生科技计划项目(No.Wsjj2017025)。

摘　　要：目的:探究1,4,5-三磷酸肌醇受体(IP3R)在促黄体素-表皮生长因子受体(LH-EGFR)信号诱导卵母细胞第一次减数分裂恢复中的作用及机制。方法:构建小鼠卵丘-卵母细胞复合物(COCs)体外培养模型以及卵泡培养模型,研究IP3R特异性抑制剂2-氨乙基二苯基硼酸脂(2-APB)及heparin对LH/EGF诱导减数分裂恢复以及EGF诱导卵丘扩展的影响,利用real-time PCR技术检测卵丘扩展因子的表达,钙离子荧光探针Fluo 3-AM检测胞内钙离子水平变化,酶联免疫法检测胞内cGMP水平变化。结果:IP3R抑制剂2-APB和heparin能够有效抑制EGF诱导的减数分裂恢复(P<0.05),同时显著降低了COCs内cGMP的含量(P<0.05);2-APB和Heparin还能够抑制EGF诱导的卵丘扩展,同时显著降低卵丘扩展因子的mRNA表达水平(P<0.05);激活IP3R可升高细胞内钙离子水平,而2-APB及heparin能够有效降低EGF升高的卵丘细胞内钙离子水平(P<0.05);利用卵泡培养模型进一步的研究表明,2-APB及heparin同样显著抑制了LH诱导的卵母细胞减数分裂恢复(P<0.05)。结论:LH-EGFR信号通路通过IP3R升高卵丘细胞内钙离子水平,诱导卵母细胞第一次减数分裂的恢复。AIM: To explore the effect of inositol 1,4,5-trisposphate receptor(IP3R) in luteinizing hormone-epidermal growth factor receptor(LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs(COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate(2-APB) and heparin(IP3R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP3R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation(P<0.05) and decreased cGMP levels in COCs(P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression(P<0.05). The activation of IP3R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells(P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation(P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP3R, resulting in meiotic resumption.

关 键 词：卵母细胞减数分裂 钙离子 1 4 5-三磷酸肌醇受体 LH-EGFR信号通路 

分 类 号：R363.2[医药卫生—病理学] R329.25[医药卫生—基础医学]