Cu2+对小鼠精原细胞活性氧及线粒体功能的影响  

Effects of Cu2+ on reactive oxygen species and mitochondrial function in mouse spermatogonia

作　　者：陈汉明 康振龙 刘高阳[1] 庞聪颖 曾绮雯[1] 刘冰贤 李英[1] CHEN Han-ming;KANG Zhen-long;LIU Gao-yang;PANG Cong-ying;ZENG Qi-wen;LIU Bing-xian;LI Ying(College of Veterinary Medicine.South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院

出　　处：《中国兽医科学》2019年第12期1586-1593,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD0502203)

摘　　要：通过终浓度含Cu^2+为0μmol/L(对照组)、10μmol/L(低浓度组)、50μmol/L(中浓度组)、100mol/L(高浓度组)的体外培养体系分别处理小鼠精原细胞(GC-1 spg细胞),在培养12、24、36 h时,应用流式细胞仪检测GC-1 spg细胞的活性氧(ROS)水平及线粒体膜电位(Δψm)变化,应用高分辨率呼吸仪检测基础呼吸、质子漏、呼吸控制比率(RCR)、剩余氧通量(ROX),以评价不同浓度Cu^2+对体外培养GC-1 spg细胞ROS产生及线粒体功能的影响。结果显示:与对照组相比,培养12 h三个Cu^2+处理组的ROS水平显著升高,低浓度组和中浓度组的ROX显著升高,Δψm、基础呼吸、质子漏和RCR无明显变化;培养24 h中浓度组和高浓度组的ROS和ROX显著升高,三个Cu^2+处理组Δψm、中浓度组和高浓度组的基础呼吸和高浓度组的RCR显著下降,质子漏无明显变化;培养36 h中浓度组和高浓度组ROS水平显著升高、三个Cu^2+处理组的质子漏、ROX显著升高,三个Cu^2+处理组的Δψm、中浓度组和高浓度组的RCR显著下降,基础呼吸无明显变化。结果表明:在Cu^2+处理的GC-1 spg细胞中,ROS的产生先于Cu^2+诱导的线粒体功能障碍。To investigate the effects of different concentrations of Cu^2+on reactive oxygen species(ROS) production and mitochondrial function in GC-1 spg cells,the GC-1 spg cells were exposed to different concentrations of Cu^2+,respectively:0 μmol/L(control group),10 μmol/L(low-dose group),50 mol/L(medium-dose group),100 μmol/L(high-dose group).After treating for 12,24 and 36 h, the ROS production and the change of mitochondrial membrane potential (Δψm)were determined by flow cytometry(FCM), and the routine respiration, proton leak, respiratory control ratio(RCR),residual oxygen consumption(ROX) were assayed by high-resolution respirometry.The results showed that the ROS level in three Cu^2+treatment groups, the ROX in low-dose group and medium-dose group were significantly increased compared to the control group, whereas the Δψm, routine respiration, proton leak, and the RCR were inconspicuous among the four groups at 12 h.The ROS and the ROX in medium-dose group and high-dose group were significantly increased,and the Δψm in three Cu^2+treatment groups,the routine respiration in medium-dose group and high-dose groups,the RCR in high-dose group were significantly decreased compared to the control group,but there was no significant change in proton leak between the four groups at 24 h. Compared to the control group, the ROS in medium-dose group and high-dose group, the proton leak and the ROX in three Cu^2+treatment groups were significantly increased, the Δψm in three Cu^2+ treatment groups, the RCR in medium-dose group and high-dose group were significantly decreased, but routine respiration had no obvious change at 36 h.These results confirmed that ROS production in Cu^2+ treated GC-1 spg cells preceded Cu^2+-induced mitochondrial dysfunction.

关 键 词：CU^2+ GC-1 spg细胞 线粒体功能 活性氧 线粒体膜电位 

分 类 号：S859.8[农业科学—临床兽医学]