精子特异性钙通道CatSper1对精子线粒体呼吸与能量代谢的影响

The Effects of CatSper1 on Mitochondrial Respiration and Energy Metabolism in Sperm

目的:应用Cat Sper1单克隆抗体抑制Cat Sper1的功能,检测精子线粒体呼吸功能及能量合成能力,观察Cat Sper1对精子线粒体呼吸与能量代谢的影响。方法:健康志愿者20例,检测精液均符合WHO健康标准。手淫法获取精液,经过上游法处理后每份精液分为A、B两组,分别与Earles液以及50μg/m L抗Cat Sper1多克隆抗体共孵育。在1 h,2 h,4 h后分别检测两组精子精液参数、细胞线粒体呼吸控制率RCR及精子细胞ATP含量。结果:与A组比较,B组精子各时间点a+b(%)尤其是a(%)均明显下降,差异具有统计学意义(P<0.01);1 h后B组精子a(%)即明显下降,2 h,4 h后精子下降缓慢,与1 h相比无明显差异,无统计学意义(P>0.05)。与A组比较,B组精子各时间点态3值、呼吸控制率RCR以及精子细胞ATP含量均明显下降,差异具有统计学意义(P<0.01);B组精子中,2 h、4 h节点精子态3值、呼吸控制率RCR以及精子细胞ATP含量与1 h无明显差异,无统计学意义(P>0.05)。结论:阻断精子特异性钙通道Cat Sper1,可降低精子细胞线粒体呼吸功能,减少精子生成ATP的能力,从而使精子活力降低。为精索静脉曲张引起精子Cat Sper1表达下降,从而导致不育的机制寻找可能的理论依据。

Objective： To observe the effects of Cat Sper1 on sperm mitochondrial respiration and energy metabolism by examining RCR and ATP after the inhibition of Cat Sper1 by Cat Sper1 monoclonal antibodies. Methods： The semen from 20 volunteers were conforming to standard of WHO, and were available by masturbation. Each sample was individually processed using a swim-up procedure and were divided into two groups（group A,B）. The sample in group A were incubated with Earles, while the sample in group B were respectively incubated with anti Cat Sper1 monoclonal antibodies at a concentration of 50 μg/m L. We measured the semen parameters,Cells mitochondrial respiratory control rate（RCR） and sperm ATP content. Results： Compared with group A, the percentage of sperm motility with grade a＋b, especially grade a in sperms was significantly decreased in group B at all time point（P〈0.01）. It becomes small of the descent rate of a（%） at the time of 2 h, 4 h, and there were no difference in a（%） among the time point in group B（P〉0.05）. The state III, RCR and ATP content in sperm were significantly decreased in group B than that in group A（P〈0.01）. But there were no difference in state III, RCR and ATP content in sperm among the time point of 1 h, 2 h and 4 h in group B. Conclusion： Inhibition of Cat Sper1 can reduce the sperm vitality via hindering cells mitochondrial respiratory function and decreasing the ability to generate ATP in sperm.The paper might find a new theory for the decrease of Cat Sper1 in sperm and infertility caused by varicocele.