色瑞替尼对精子制动作用和质膜损伤的体外研究

发布时间：2018-09-10 11:33

【摘要】：目的研究前期筛选到的精子功能调节候选化合物色瑞替尼的体外精子制动作用和机制。方法计算机辅助精液分析(computer assistant sperm analysis,CASA)系统测定色瑞替尼对人和小鼠高活力精子20 s内的制动效果;低渗膨胀(hypo-osmotic swelling,HOS)实验、SYBR-14/PI染色探究精子质膜的完整性;用电镜观察色瑞替尼处理后精子质膜的变化;用色瑞替尼处理VK2/E6E7、End1/E6E7、Ect1/E6E7细胞,CCK-8及荧光染色法检测细胞活性。结果色瑞替尼对于(5~10)×106/mL人精子的20 s瞬间杀精最小有效浓度(minimal effective concentration,MEC)为(74.87±31.46)μmol/L,而相同条件下测到的杀精药物壬苯醇醚(nonoxynol-9,N-9)的MEC为(219.75±20.89)μmol/L。CASA系统分析表明,色瑞替尼对精子的制动作用有时间-浓度依赖关系。HOS实验及SYBR-14/PI染色法验证色瑞替尼对精子质膜有一定的损伤,并可致精子死亡。电镜显示色瑞替尼处理后的精子质膜破裂,头尾分离较多。色瑞替尼对上皮细胞有一定损伤。结论色瑞替尼对精子有快速的制动作用,具有潜在的避孕药效。
[Abstract]:Objective to study the in vitro sperm immobilization and mechanism of Seritinib, a candidate compound for spermatozoa regulation. Methods computer assisted semen analysis (computer assistant sperm analysis,CASA) system was used to determine the immobilization effect of seritinib on human and mouse hyperactive spermatozoa for 20 seconds, and hypotonic expansion (hypo-osmotic swelling,HOS) test was used to study the integrity of sperm plasma membrane by SYBR-14 / Pi staining. The changes of plasma membrane of spermatozoa treated with seritinib were observed by electron microscope, CCK-8 of VK2/E6E7,End1/E6E7,Ect1/E6E7 cells treated with seritinib and the activity of spermatozoa were detected by fluorescence staining. Results the minimal effective concentration (minimal effective concentration,MEC) of semitinib for (5 ~ 10) 脳 106/mL human sperm in 20 s was (74.87 卤31.46) 渭 mol/L, while the MEC of nonylphenyl alcohol ether (nonoxynol-9,N-9) was (219.75 卤20.89) 渭 mol/L.CASA under the same conditions. The effect of seritinib on the immobilization of spermatozoa was time-concentration-dependent. HOS test and SYBR-14/PI staining showed that Seritinib could damage the plasma membrane of spermatozoa and cause sperm death. Electron microscope showed that the plasma membrane of spermatozoa was ruptured and the separation of head and tail was more after treatment with seritinib. Seritinib has some damage to epithelial cells. Conclusion Seritinib has a rapid immobilization effect on spermatozoa and has potential contraceptive effect.
【作者单位】：复旦大学上海医学院生殖生物学研究室;复旦大学生殖与发育研究院上海市计划生育科学研究所国家人口和计划生育委员会计划生育药具重点实验室;
【基金】：国家重点研发计划(2016YFC1000905) 国家自然科学基金面上项目(81671508) 上海市科技创新行动计划项目(15431903000)~~
【分类号】：R698.2

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