孕酮通过ABHD2受体介导的cAMP-PKA信号通路调控精子受精功能的研究  

Study on the progesterone regulating sperm fertilization through ABHD2 receptormediated cAMP-PKA signaling pathway

作　　者：江峰[1] 朱勇 陈颖[1] 汤晓峰[1] 杨岳洲 陈国武[1] 刘玉林[1] 孙晓溪[1] Jiang Feng;Zhu Yong;Chen Ying;Tang Xiaofeng;Yang Yuezhou;Chen Guowu;Liu Yulin;Sun Xiaoxi(Shanghai JIAI Genetics&IVF Institute-China USA Ceter,200011,China;Obstetrics and Gynecology Hospital of Fudan University,Shanghai,China)

机构地区：[1]上海集爱遗传与不育诊疗中心,上海200011 [2]复旦大学附属妇产科医院

出　　处：《中国男科学杂志》2020年第6期12-16,共5页Chinese Journal of Andrology

基　　金：上海市卫计委科研课题(项目编号:201740158);上海市市级科技重大专项"国际人类表型组计划(一期)"项目资助(项目编号:2017SHZDZX01)。

摘　　要：目的研究ABHD2在不育症患者精子膜表面的表达差异,探究孕酮是否通过ABHD2调控cAMP-PKA进而影响精子的功能活性。方法收集男性正常生育组精子、少弱精子组和顶体异常组精子样本(各30例),通过RT-PCR、蛋白Western-Blot印迹等方法分析转录水平及蛋白表达的差异,且采用体外添加孕酮、ABHD2抗体、PKA拟制剂H-89,选择观测正常生育组(对照组)与少弱精子组(实验组)男性精子ABHD2的蛋白表达水平,以及cAMP、PKA浓度等变化。结果少弱精子组和顶体异常组中ABHD2 m RNA转录水平以及ABHD2蛋白表达水平显著减少,具有统计学意义(F:90.84,P<0.01);在正常生育组中加入孕酮处理后,ABHD2蛋白水平表达明显增加;而少弱精子组中加入孕酮处理后,与未加孕酮相比,ABHD2蛋白水平表达增加不明显。进一步研究,在正常生育组中加入孕酮(P4)处理后,cAMP、PKA浓度极显著增加(P<0.01),继续加入ABHD2抗体或PKA拟制剂H89后,cAMP、PKA浓度均显著减少(P<0.01)。结论临床不育与正常生育男性精子之间ABHD2的转录和表达存在差异,孕酮通过ABHD2受体介导的cAMP-PKA信号通路调控精子受精功能。Objective To explore the expression of ABHD2 in the sperm membrane surface,and clarify whether ABHD2-mediated cAMP-PKA signaling pathway participates in progesterone regulating sperm function in infertile patients.Methods Semen samples from healthy,patients with oligoasthenospermia and the patients with abnormal acrosome reactions were collected,30 samples of each group.Their transcription and protein expression were comparatively analyzed.The protein expression level of ABHD2 and the changes in the concentration of cAMP and PKA in the normal fertile group(the control group)and the oligoasthenospermia group(the experimental group)were detected after treating with progesterone,ABHD2 antibody,or PKA inhibitor H-89.Results The transcription and protein expression levels of ABHD2 in the oligoasthenospermia group and the group of acrosome abnormality were all significantly lower than those of the normal group(F:90.84,P<0.01).After adding progesterone to the fertile group,the protein expression of ABHD2 in the fertile group was increased significantly.But there was no obvious increase in ABHD2 protein expression in the oligoasthenospermia group.Further research showed that the concentrations of cAMP and PKA were increased extremely significantly in the normal fertile group after treated with progesterone(P<0.01),while the concentration of cAMP and PKA were decreased significantly in the fertile group after treated with ABHD2 antibody or PKA inhibitor H-89(P<0.01).Conclusion There are differences in ABHD2 m RNA and protein expressions in sperm between males with clinical infertility and normal fertile males.The progesterone can regulate sperm fertilization through ABHD2 receptor-mediated cAMP-PKA signaling pathway.

关 键 词：精子 孕酮 ABHD2蛋白表达 cAMP-PKA信号通路 

分 类 号：R349.54[医药卫生—基础医学] R321.2