rhTNF-a对人正常精子超微结构的实验研究

Electron Microscopic Technique to Assess Effect of rhTNF-a on Ultra Structure of Human

目的观察rhTNF-a在体外对人正常精子超微结构的影响。方法26例健康成年男性手淫法取得精液,percoll梯度离心法得到的精子为精子模型,将精子浓度调整为10×106/mL,在37℃、5%CO2的环境中,分别与终浓度为60、90、270pg/mL的rhTNF-a孵育,同时设立对照组。在0.5h和4h时间段取样。TEM观察试验组和对照组精子的超微结构。结果实验组与对照组相比较60、90、270 pg/mL的超微结构发生不同程度的变化:细胞膜变薄,扭曲或成波浪状改变,大部分精子细胞膜不连续,细胞膜与顶体外膜间隙增宽,透亮度增强,顶体变厚,甚至可见顶体外膜破裂,顶体内容物流失;线粒体密度降低,透光度增强,严重者线粒体外形变圆,内有空泡形成;核膜与核内染色质未发生改变;纤维轴丝9+2模式未改变。结论rhTNF-a可破坏精子的顶体和膜结构及线粒体结构而降低精子受精能力。

Objective To study the effect of rhTNF- a on ultrastructure of health human sperm in vitro. Metheds Fresh semen samples were collected and analyzed according to the protocols by masturbation in 26 healthy adult men. Semen samples from 16 health men were prepared using percoll centrifugation, the sperm suspension was diluted to a concentration of l0 × 10^6/mL in Ham＇s F10 medium and incubated with rhTNF -a solution （final concentration 60,90,270pg/ mL,respectively ）for 0.5h and 4h at 37℃ in 5%CO2, and comparative studies were made with a control group. Samples were scanned by TEM. Results ultrastructure of health human sperm in experiment groups （final concentrate ion 60, 90 and 7.70 pg/mL） and that in control groups have significant differences. The segmental breakages of acrosome with a show of atypical vacuoles are contents of low electronic density. The acrosome is dilated and disrupted. The spaces widened with spillover material in between are discernible on the border. The plasma membrane is impaired. Conclusion. rhTNF - α can destroy the ultrastructure of the health human sperm in vitro.