线粒体氧化磷酸化抑制剂FCCP体外对人精子动力学和线粒体功能的影响  

Influence of FCCP,inhibitor of mitochondrial oxidative phosphorylation,on motility and mitochondrial function of human spermatozoa in vitro

作　　者：许剑锋 王晓尉 张林媛[2] 傅龙龙 梁小薇 XU Jian-feng;WANG Xiao-wei;ZHANG Lin-yuan;FU Long-long;LIANG Xiao-wei(Department of Male Clinical Research/Human Sperm Bank,National Research Institute for Family Planning,Beijing100081;National Institute for Occupational Health & Poison Control,China CDC,Beijing100050)

机构地区：[1]国家卫生健康委科学技术研究所男性临床研究室,北京100081 [2]中国疾病预防控制中心职业卫生与中毒控制所毒理室,北京100050

出　　处：《生殖医学杂志》2019年第10期1232-1237,共6页Journal of Reproductive Medicine

基　　金：中华医学会临床医学科研专项资金-生殖医学青年医师研究与发展项目(18010310760)

摘　　要：目的通过线粒体氧化磷酸化(OXPHOS)特异性抑制剂FCCP对人精子活动力及其线粒体功能影响的研究,探讨氧化磷酸化在精子能量代谢中的作用。方法选择来自捐精志愿者的正常精液8份,优选后制备精子悬液,将每份精子悬液分为4组,分别与终浓度为0μmol/L(对照组)、2.5μmol/L、5μmol/L和10μmol/L的FCCP共孵育1h、3h、5h,以精子动力学参数、线粒体膜电位、精子细胞内ATP含量、精子质膜完整性作为评价指标,分析比较各组间的差异。结果(1)各组精子活动率和其他各项运动参数随着FCCP浓度增高呈下降趋势:孵育1h后,与对照组相比,仅10μmol/L组的精子活动率、前向运动百分率和精子头侧摆幅度(ALH)显著下降(P<0.05),其余指标无显著性变化,而其余浓度处理组的各指标变化均无统计学意义(P>0.05);孵育3h后,10μmol/L组的精子活动率、前向运动百分率、平均路径速率(VAP)、直线速率(VSL)、曲线速率(VCL)、ALH和鞭打频率(BCF)均显著降低(P<0.05),5μmol/L组的ALH和BCF指标显著下降(P<0.05);孵育5h后,与对照组相比,10μmol/L组的前述指标继续显著性下降(P<0.05),5μmol/L组的精子活动率和前向运动百分率也出现显著降低(P<0.05),而2.5μmol/L组各指标差异均无统计学意义(P>0.05)。(2)精子线粒体膜电位(MMP)和ATP含量随FCCP浓度增加逐渐降低,10μmol/L组的MMP和ATP含量显著低于对照组(P<0.05)。(3)质膜完整性比较中,10μmol/L组比对照组显著降低(73.94%vs.84.53%)(P<0.05)。(4)随着FCCP孵育时间延长,各组精子活动率和前向运动百分率呈下降趋势。结论不同浓度的FCCP体外处理精子后,精子活动率和其他各项运动参数,以及反映线粒体活性的线粒体膜电位和ATP含量呈浓度依赖性下降,FCCP所抑制的线粒体氧化磷酸化是精子能量代谢的重要途径。Objective:To investigate the effects of specific inhibitor FCCP for mitochondrial oxidative phosphorylation(OXPHOS)on sperm motility and mitochondrial function.Methods:Eight normal semen samples from volunteers were purified by density gradient centrifugation and suspended in BWW medium.Each sample was divided into 4 parts and the suspensions were incubated with FCCP in 0,2.5,5 and 10μmol/L for 1,3,5 hours respectively.Sperm motility parameters,mitochondrial membrane potential,ATP content in sperm cells and sperm plasma membrane integrity as evaluation indicators were analyzed and compared among the groups.Results:Sperm motility and other motile parameters declined with the increased concentration of FCCP.After incubation for 1 hour,the sperm motility,progressive motility and sperm amplitude of lateral head displacement(ALH)decreased significantly in the 10μmol/L group compared with the control group(P<0.05),while there was no significant difference in the other indicators.There was no significant change in each indicator of the other concentration of FCCP treatment groups(P>0.05).After incubation for 3 hours,sperm motility,progressive motility,average path velocity(VAP),straight-linear velocity(VSL),curvilinear velocity(VCL),ALH and beat-cross frequency(BCF)were significantly decreased in the 10μmol/L group(P<0.05),while only ALH and BCF were significantly decreased in the 5μmol/L group(P<0.05).After 5 hours of incubation,the aforementioned indicators in the 10μmol/L group continued to decrease significantly compared with the control group(P<0.05),and the sperm motility and progressive motility in the 5μmol/L group also decreased significantly(P<0.05).There was no significant difference in the each indicator in the 2.5μmol/L group(P>0.05).The mitochondrial membrane potential(MMP)and ATP content of sperm decreased gradually with the increase of FCCP concentration,and the MMP and ATP levels in the 10μmol/L group were significantly lower than those in the control group(P<0.05).The plasma membrane integrity

关 键 词：精子 精子活动力 线粒体 氧化磷酸化 

分 类 号：R339.2.1[医药卫生—人体生理学]