Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa
Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa
复制标题
DOI：
10.1210/jc.2007-2616
复制
发表时间：
2008-08-01
影响因子：
5.8
通讯作者：
Aitken, R. John
中科院分区：
文献类型：
--
作者：
Koppers, Adam J.;De Iuliis, Geoffry N.;Aitken, R. John
背景：男性不育与有缺陷的精子产生过多的活性氧（ROS）有关。然而，这种活动的亚细胞起源尚不清楚。目的：本研究的目的是确定精子线粒体在产生与精子功能缺陷相关的氧化应激中的重要性。方法：使用荧光探针MitoSOX red和BODIPY C-11结合流式细胞术检测细胞内线粒体ROS生成和脂质过氧化。对精子运动的影响通过计算机辅助精子分析来测量。结果：人类精子中线粒体电子传输流的中断导致复合物I（鱼藤酮敏感）或III（粘噻唑，抗霉素A敏感）通过独立于线粒体膜电位的机制产生ROS。在络合物III处激活ROS生成导致过氧化氢快速释放到细胞外空间，但没有检测到过氧化损伤。相反，在复合体I的线粒体内膜基质侧诱导ROS导致中部的过氧化损伤和精子运动的丧失，这可以通过同时存在的α -生育酚来防止。有缺陷的人精子自发产生线粒体ROS，其方式与运动性呈负相关。用二氢乙锭同时测量一般细胞ROS生成表明，这种测量中68%的变异性可以用线粒体ROS生成的差异来解释。结论：精子线粒体在缺陷精子的氧化应激中起着重要作用。
Context: Male infertility has been linked with the excessive generation of reactive oxygen species (ROS) by defective spermatozoa. However, the subcellular origins of this activity are unclear.Objective: The objective of this study was to determine the importance of sperm mitochondria in creating the oxidative stress associated with defective sperm function.Method: Intracellular measurement of mitochondrial ROS generation and lipid peroxidation was performed using the fluorescent probes MitoSOX red and BODIPY C-11 in conjunction with flow cytometry. Effects on sperm movement were measured by computer-assisted sperm analysis.Results: Disruption of mitochondrial electron transport flow in human spermatozoa resulted in generation of ROS from complex I (rotenone sensitive) or III (myxothiazol, antimycin A sensitive) via mechanisms that were independent of mitochondrial membrane potential. Activation of ROS generation at complex III led to the rapid release of hydrogen peroxide into the extracellular space, but no detectable peroxidative damage. Conversely, the induction of ROS on the matrix side of the inner mitochondrial membrane at complex I resulted in peroxidative damage to the midpiece and a loss of sperm movement that could be prevented by the concomitant presence of alpha-tocopherol. Defective human spermatozoa spontaneously generated mitochondrial ROS in a manner that was negatively correlated with motility. Simultaneous measurement of general cellular ROS generation with dihydroethidium indicated that 68% of the variability in such measurements could be explained by differences in mitochondrial ROS production.Conclusion: We conclude that the sperm mitochondria make a significant contribution to the oxidative stress experienced by defective human spermatozoa.