线粒体及其自噬与不同程度弱精子症的关系*

Correlation between mitochondria and mitophagy in different degrees of Asthenospermia*

摘要:

目的 观察弱精子症患者的射精精子中是否存在自噬及线粒体自噬现象，并探讨线粒体及其自噬对精子活动力的影响。方法 收集湖北医药学院附属人民医院男科及生殖中心就诊的精液样本，并依照计算机辅助精液分析系统（CASA）检测结果，分为正常对照组和轻、中、重度弱精子症组，每组30例。采用电镜观察各组精子线粒体形态及自噬小体分布；Western blotting检测精子线粒体膜蛋白及自噬标志性蛋白TOM20、LC3等的表达水平；共聚焦显微镜检测精子中的LC3及线粒体荧光斑点，以了解精子中发生的自噬是否属于特异性线粒体自噬。结果 不同程度弱精子症患者的精子中，线粒体分布及自噬发生程度均有不同的表现，两者存在共定位的特点，并且与弱精子症程度呈负相关（P?<0.05）。结论 线粒体自噬是人类精子的活性过程，自噬的活化在人类精子生理活动中起积极作用，保持并调节精子活动力。

Abstract:

Objective To observe the phenomenon of autophagy and mitophaghy of ejaculated sperm in the male patients with asthenospermia (AZS), and to determine the role of mitochondria and their autophagy among patients with different level of AZS in China. Methods According to the WHO manual (the 5th edition), published in 2010, semen samples were collected from the Andrology clinics in the People’s hospital which were affiliated to Hubei University of Medicine, and were divided into 4 group, the normal control group, mild group, moderate group and severe group (30 cases in each group). Electron microscope detection was performed to analyze the number and morphology of mitochondria and the distribution of autophagosomes in the sperm of each group. Western blotting method was used to judge the expression of TOM20, LC3 and other proteins related to mitochondrial autophagy. LC3 and mitochondrial fluorescence spots in sperm were detected by confocal laser microscopy to exam if autophagy in sperm is the idiopathic mitochondrial autophagy. Results In different degree of asthenospermia, the distribution of mitochondria and the degree of autophagy were different, they had the characteristics of CO location, and they were negatively correlated with the degree of asthenospermia (P?