prohibitin表达与男性不育的相关性研究 - fdurop - 复旦大学.docVIP

Prohibitin表达与男性不育的相关性研究 复旦大学上海医学院 区嘉贤、黄思菲 指导老师 陈红 摘要：【目的】Prohibitin为高度保守的精子线粒体膜蛋白。体细胞Prohibitin表达缺失与线粒体膜去极化及氧自由基生成增加相关。已发现人类精液中氧化剂水平升高或导致男性不育，而精子活力是氧化损伤最早、最敏感的指标。基于Prohibitin在线粒体中具分区及协助呼吸链组装的功能，分别检测少精症与弱精症患者的精子线粒体膜电位与Prohibitin表达以阐明Prohibitin表达与精子线粒体膜电位、精子活力与男性生育能力的关系。【方法】收集68位30-40岁、拟行ICSI/IVF的男性精液样本，按2010年WHO标准分为正常、少精症与弱精症，分别进行精子浓度、活力与形态检测。活精子经MitoTracker Red CMXRos probe染色及流式细胞技术测定线粒体膜电位，固定后行免疫荧光检测Prohibitin与线粒体共定位情况，另收集样本行Western印迹检测精子线粒体内Prohibitin表达浓度。实验结果经单向 ANOVA与Newman-Keuls post-test分析；相关性通过Spearman秩相关系数和线性回归模型分析。【结果】按2010年WHO标准，正常、少精症与弱精症样本数分别为37、23与8。少精症（49.58±4.0；P0.05）与弱精症患者（26.91±5.8；P0.01）的线粒体膜电位较正常男性（63.23 ± 3.0）显著降低，当中弱精症患者的下降幅度大于少精症患者（P0.01）；少精症与弱精症患者的精子Prohibitin表达也较正常男性显著下降（P0.05）。统计学分析证实：正常、少精症与弱精症男性的精子Prohibitin表达、线粒体膜电位及精子活力呈正相关（P0.05）。【结论】Prohibitin或通过改变线粒体膜电位与提高氧自由基水平影响精子活力，进一步提示Prohibitin表达可作为不育男性精子质量的指标。 关键词：临床医学；线粒体；生物标志物；诊断学 ABSTRACT: Prohibitin is a highly conserved major sperm mitochondrial membrane protein whose absence in somatic cells is associated with mitochondrial membrane depolarization and increased generation of reactive oxygen species (ROS). Our recent findings suggest that high levels of oxidants in human semen may contribute to male infertility and that sperm motility could be the earliest and most sensitive indicator of oxidative damage. Based on Prohibitins roles in mitochondrial sub-compartmentalization and respiratory chain assembly, we examine sperm Prohibitin expression and mitochondrial membrane potential in infertile males with poor sperm motility (asthenospermia, A) and/or low sperm concentrations (oligoasthenospermia, OA). Here we demonstrate that mitochondrial membrane potential is significantly lower in sperm from A and OA subjects than in normospermic (N) subjects; the decrease is more severe for OA than for A subjects. Prohibitin expression is also significantly lower in sperm from A and OA subjects. Importantly, significantly positive correlations are found among Prohibitin expression, mitochondrial membrane