【摘要】 目的:观察rhTNF-a在体外对人正常精子线粒体功能和质膜完整性的影响。方法:40例健康自愿者正常精液,Peroll梯度离心将精子密度调整为10×106/mL,分别与终浓度0 pg/mL(对照组)、30 pg/mL、60 pg/mL、90 pg/mL、270 pg/mL的rhTNF-a孵育。在0.5 h、1 h、2 h、3 h、4 h时间段取样,用终浓度为10 mg/mL 的Rh123和PI双染色精子,流式细胞仪检测精子线粒体膜电位及质膜完整性的变化。结果:与对照组相比较,60 pg/mL、90 pg/mL、270 pg/mL各组线粒体功能良好的精子明显减少,这种变化趋势与TNF-a浓度和孵育时间呈正相关(P <0.01)。30 pg/mL组的精子线粒体功能与对照组相比无统计学差异(P >0.05);30 pg/mL、60 pg/mL、90 pg/mL、270 pg/mL各组的PI和Rh123双染色均阳性的精子数较对照组多,且与rhTNF-a的浓度和孵育时间呈正相关(P <0.01)。结论: rhTNF-a对人精子线粒体功能有一定的影响,提示有可能干扰精子的能量代谢,降低精子受精能力,并有可能通过线粒体凋亡途径,促进精子凋亡;同时影响精子质膜的完整性。更多还原

【Abstract】 Objective: To study in vitro effect of rhTNF-a on mitochondrial membrane potential andmembrane integrity of human sperm. Methods: Semen samples from 40 healthy men were prepared by Percollcentrifugation, and incubated with rhTNF-a solution (final concentration: 30 pg/mL, 60 pg/mL, 90 pg/mL and 270 pg/mL, respectively) for 0.5 h, 1 h, 2 h, 3 h and 4 h, while comparative studies were made with a control group. Semensamples (250 mL) were stained in the presence of 10 mg/mL Rh123 and PI, and mitochondrial membrane potentialand membrane integrity of human sperm were analyzed by flow cytometry. Results: Significant differences werefound between experimental and control groups on mitochondrial membrane potential, respectively (P<0.01)expect for final concentration 30 pg/mL group (P>0.05).The number of sperm with normal mitochondrial functiondecreased with increasing rhTNF-a concentration and incubating time. On the other hand, significant differenceswere found between experimental and control groups on membrane integrity, respectively (P<0.01). The number ofsperm with normal membrane integrity decreased with increasing rhTNF-a concentration and incubating time.Conclusion: rhTNF-a can inhibit the function of mitochondron in healthy human sperm, which might disruptsperm mitochondrial enzymatic activities. Meanwhile, rhTNF-a can damage sperm membrane integrity.更多还原