生育男性和白细胞精子症患者精子线粒体基因缺失和含量的研究  

Sperm mtDNA Content and mtDNA^(4977bp) Deletion in Normal and Leukocytospermia Men

作　　者：周万灏[1] 马旭 姜辉[3] 袁任培[3] 陈茜[3] 隋玉洁[3] 贾孟春 

机构地区：[1]中国协和医科大学研究生院,北京100730 [2]国家人口与计划生育委员会科学技术研究所内分泌室,北京100081 [3]北京大学第三临床医院男科中心,北京100083

出　　处：《中华男科学杂志》2008年第5期391-395,共5页National Journal of Andrology

基　　金：国家自然科学基金(30670784)

摘　　要：目的:测定生育男性与白细胞精子症患者精子线粒体DNA(mtDNA)含量、mtDNA4977bp缺失及精浆活性氧(ROS)水平,探讨精液中白细胞增加、ROS水平升高与精子mtDNA变化的相关性。方法:选取门诊白细胞精子症患者78例(病例组)、正常生育男性31例(对照组),采用Percoll梯度离心法将精液样本分为上清层、30%层和80%精子层,分别应用荧光实时定量PCR技术对每个精子mtDNA含量及mtDNA4977bp缺失进行定量分析,同时应用流式细胞术测定精浆ROS水平。结果:病例组精浆中ROS含量显著高于对照组(P<0.01)。与对照组比较,病例组各层精子mtDNA含量均显著升高,其P值均<0.01。在上清层和80%层,病例组mtDNA4977bp缺失与对照组差异不显著;而在30%层,病例组mtDNA4977bp缺失明显高于对照组(P<0.01)。对照组与病例组各样本中ROS含量分别与30%层精子和80%层精子的mtDNA含量间呈显著正相关(r=0.347,P<0.01;r=0.456,P<0.01),与上清层精子mtDNA含量无相关性。结论:白细胞增多和ROS升高可能是导致精子mtDNA含量显著增高的原因之一,但是对于mtDNA4977bp缺失没有显著影响。Objective ： To determine the sperm mtDNA content, mtDNA4977bp deletion and ROS in the seminal plasma of normal and leukocytospermia men, and to investigate the correlation of the changes of sperm mtDNA with the increase of leukocytes and reactive oxgygen specias （ROS） in the seminal plasma. Methods： Seventy-eight semen samples from leukocytospermia patients and 31 from healthy donors were divided into 3 layers, supematant fluid, 30% sperm and 80% sperm, by PercoU gradient centrifugation, their sperm mtDNA content and mtDNA^4977bp deletion quantitatively analyzed by real-time PCR, and the level of ROS determined by flow cytometry. Results ： The ROS in the seminal plasma and the sperm mtDNA contents of the three layers were all significantly higher in the leukocytospermia group than in the healthy control （ P 〈 0.01 ）. In the supematant fluid and 80% layers, mtDNA^4977bp deletion showed no obvious difference between the control and the leukocytospermia group, but was significantly higher in the 30% layer of the latter （P 〈 0.01 ）. The ROS level was found positively correlated with the mtDNA content in the 30% （r = 0.347, P 〈 0.01 ） and the 80% layer （ r = 0.456, P 〈 0.01 ）, but not in the supernatant layer. Conclusion ： The increase of leukocytes and ROS may be one of the causes of the enhanced sperm mtDNA content, but has no significant impact on the mtDNA^4977bp deletion.

关 键 词：白细胞精子症 活性氧 精子mtDNA含量 mtDNA^4977bp缺失 荧光实时定量PCR 

分 类 号：R698[医药卫生—泌尿科学]