TRPV1离子通道对精子线粒体膜电位的影响

The Effect of TRPV1 on Human Sperm Mitochondrial Membrane Potential

目的探讨TRPV1(transient receptor potential vanilloid type1,TRPV1)离子通道对精子线粒体膜电位(mitochondrial membrane potential,MMP)的影响。方法手淫法收集健康生育男性精液,采用上游法分离精液精子。每份精液等分为五份,分别接受不同的处理。实验分五组,即四个实验组和一个对照组。实验组分别加入20μmmol/L capsaicin(B组)、50μmmol/L capsaicin(C组)、20μmmol/L capsazepine(D组)和50μmmol/L capsazepine(E组),对照组(A组)不加药物。采用计算机辅助的精液分析(CASA)测定精子活率及运动参数,JC-1探针法检测精子线粒体膜电位的变化。结果 TRPV1配体对精子各项检测指标的影响呈现剂量和时间依赖性。CASA检测显示,TRPV1配体作用30分钟后精子活率明显下降,各实验组精子活率分别为(53.60±8.05)%、(43.38±8.66)%、(48.32±8.20)%及(35.42±6.21)%,与对照组比较差异均有统计学意义(P<0.01)。实验组同一药物不同剂量之间比较,差异均有统计学意义(P<0.01)。此外,除BCF和ALH外,精子各项运动参数均出现有统计学意义的改变(vs control,P<0.01),同一药物不同剂量之间亦有统计学差异(P<0.01)。各组精子线粒体膜电位检测发现,与对照组比较,实验组荧光强度下降明显(P<0.01),并且同一药物不同剂量之间比较,荧光强度变化均有统计学差异(P<0.01)。结论 TRPV1配体可能通过与TRPV1受体结合或者非受体途径,作用于精子线粒体,降低线粒体膜电位,损害精子线粒体功能,最终影响精子存活及运动功能。

Objective To investigate the effect of TRPV1 on sperm mitochondrial membrane potential.Methods Semen samples of health reproductive men were collected by masturbation,the sperm of semen separated using upper swimming method.Each portion of semen was divided into five groups,receiving different treatments respectively.The experiment was divided into five groups,four experimen tal groups and a control group.Experimental group separately were added 20 μmmol /L capsaicin（B group）,50 μmmol /L capsaicin（C group）,20 μmmol /L capsazepine（D group） and 50 μmmol /L capsazepine（E group）,the control group（A group） were not added with drugs.The sperm viability and motion parameters were determined by computer-aided semen analysis（CASA）,the changes of sperm mito chondrial membrane potential were detected by JC-1 probe method.Results The results showed that TRPV1 ligands acted on the sperm in dose and time-dependent manner.CASA detection showed live sperm rate significantly decreased after TRPV1 ligands played 30 minutes.In all experimental groups sperm motility were（53.60 ± 8.05）%,（43.38 ± 8.66）%,（48.32 ± 8.20）% and（35.42 ± 6.21）% respectively,which had statistical significance compared with the control group（P 0.01）.There was statistical significance between different dosage of the same a gent in ex perimental group（P 0.01）.Except BCF and ALH,it showed statistically significant changes in the sperm motility parameters（vs control,P 0.01）,it had also significant difference in different doses of the same drug（P 0.01）.Sperm mitochondrial membrane potential detection found that in experimental groups it decreased significantly compared with the control group（P 0.01）,the changes of fluores cence intensity had significant differ ence among different dos es of the same drugs.Conclusions With TRPV1 receptor-binding or non-receptor pathway,TRPV1 ligands act on the sperm mitochondria,reduce mitochondrial mitochondria membrane potential,damage sperm mitochondrial function,and ultimately affect sperm viability and motility.