精子的氧化损伤与线粒体解偶联蛋白2(UCP2)表达的关联性  

Relationship betweenoxidative damage of human sperm and uncoupling protein 2 expression

作　　者：王晓娜[1] 朱春芳[2] 张家燕[1] 朱楠[1] 陈华[2] 张帆[2] 葛红山[2,1] Wang Xiaona Zhu Chunfang Zhang Jiayan et al(The Second Clinical College of Wenzhou Medical University, Zhejiang 325000, Chin)

机构地区：[1]温州医科大学附属第二医院 [2]温州医科大学第二临床学院,325000

出　　处：《医学研究杂志》2017年第7期62-66,共5页Journal of Medical Research

基　　金：浙江省自然科学基金资助项目(LQ15H040006);浙江省科技厅公益性研究项目(2014C33162);浙江省教育厅项目(Y201327892);浙江省计划生育委员会项目(2014KYB345);温州市科技局项目(Y20140479,Y20140615)

摘　　要：目的通过研究过氧化氢(hydrogen peroxide,H_2O_2)处理对精子活性氧的(reactive oxygen species,ROS)产生,精子运动参数,精子DNA碎片化程度及UCP2蛋白表达的影响,探讨精子的氧化损伤与UCP2蛋白表达的相关性。方法采用密度梯度离心法处理精液,选取离心后的沉淀精子为研究对象,以终浓度分别为50μmol/L,100μmol/L和200μmol/L的过氧化氢处理精子~1H,同时设立对照组(0μmol/L)。流式细胞仪检测各组精子活性氧(ROS)含量;计算机辅助精液分析系统(CASA)分析各组精子运动参数;精子染色质结构分析试验(SCSA)检测精子DNA碎片化程度;Western blot法检测各组UCP2蛋白的表达情况。分析精子的氧化损伤与UCP2表达的关联性。结果随着过氧化氢作用浓度的增加,各处理组的ROS含量,DNA碎片化程度和UCP2蛋白的表达均存在增加,且与对照组比较,差异均有统计学意义(P<0.05);同时各处理组精子运动参数等均呈下降趋势,尤其是100μmol/L和200μmol/L组,除侧摆幅度参数外,其他参数与对照组比较,差异均有统计学意义(P<0.05)。结论过氧化氢作用后对精子造成氧化损伤(ROS升高),导致精子运动参数下降和DNA碎片化程度增加,对应的UCP2蛋白表达的增加,证明精子的氧化损伤与UCP2蛋白的表达存在关联性,提示UCP2可能参与精子体内的抗氧化作用。Objective To study the effect of hydrogen peroxide on production of reactive oxygen species（ROS） , sperm motile parameters ,DNA integrity and the level of UCP2 protein in sperm, and investigate the relationship between oxidative damage of human sperm and UCP2 protein expression. Methods High energy sperm was isolated from semen by density gradient centrifugation method. After passage, high energy sperm was treated by the final concentrations of 50μmol/L, 100μmol/L and 200p, mol/L H2O2 for t n, respectively. Meanwhile, the control group was established. The content of ROS was detected by flow cytometry, sperm motile parameters by Computer- aided sperm analysis （CASA）, DNA integrity by sperm chromatin structure assay （SCSA）, UCP2 protein expression hy Western blot. Results Compared with the control group, ROS production and DNA integrity of treatment groups were significantly in- creased（ P 〈 0.05 ）. The level of UCP2 protein expression was up - regulated while the level as concentrations of hydrogen peroxide increased（P 〈0.05）. Exposure to hydrogen peroxide, sperm motile parameters appeared a decreasing trend. Especially, High concentrations （100μmol/L and 200p, mol/L） resulted in most motile parameters （except ALH） statistically significant difference from the control group（P 〈0. 05）. Conclusion Oxidative damage caused by hydrogen peroxide, so sperm motile parameters decreased and DNA integrity increased . Hydrogen peroxide induced the increase of sperm protein UCP2 expression. It proves that the correlation between oxidative damage of human sperm and UCP2 expression and suggests that UCP2 may play a protective role on antioxidant capability of human sperm.

关 键 词：精子 过氧化氢 UCP2蛋白 氧化损伤 

分 类 号：R3[医药卫生—基础医学]