钙离子通道蛋白CatSper在吸烟小鼠模型中的表达  

Expression of sperm cation channel CatSper in smoking mice model

作　　者：姚丽娟[1] 郑圣霞[1] 吴双正[1] 孙晓科[1] 周桂香[1] 周明[1] 

机构地区：[1]安徽医科大学附属省立医院,合肥市230001

出　　处：《实用医学杂志》2014年第3期357-360,共4页The Journal of Practical Medicine

基　　金：安徽医科大学校科研基金资助项目(编号:2011xkj078)

摘　　要：目的:建立吸烟小鼠模型,检测小鼠精子特异性钙离子通道蛋白CatSper在RNA和蛋白水平表达的变化,分析与精子活力之间的关系,为吸烟导致雄性生殖损害的分子机制研究提供可靠的实验数据。方法:建立吸烟小鼠模型,运用非连续密度梯度离心分离精子并进行活力检测,RT-PCR检测小鼠精子mRNA表达的变化,免疫组织细胞化学技术观察小鼠CatSper蛋白的表达,免疫印迹进一步对CatSper蛋白表达的变化趋势进行统计学分析。结果:正常对照组,轻、中和重度吸烟组(a+b)级精子百分率[(87.6±16.2)%vs(72.9±13.0)%vs(68.1±16.0)%vs(44.6±15.0)%,P<0.05]、CatSper mRNA相对表达量(0.915±0.202 vs 0.712±0.121 vs 0.522±0.126 vs 0.258±0.177,P<0.05)、免疫印迹相对表达量(0.882±0.208 vs0.693±0.202 vs 0.471±0.251 vs 0.263±0.198,P<0.05)均依次降低,免疫组化结果显示吸烟使CatSper蛋白在睾丸组织中表达降低;相关性分析结果表明(a+b)级精子百分率与CatSper mRNA和蛋白呈正相关。结论:吸烟明显抑制小鼠钙离子通道蛋白CatSper的表达,精子活力与CatSper的表达呈正相关,提示CatSper表达降低可能是吸烟导致雄性精子活力损伤的分子机制之一。Objective To provide reliable experimental data for molecular mechanism research on male reproductive impairment by establishing the mice model of smoking to detect the expression of specific cation channel CatSper mRNA and protein in sperm, and analyze its relation with sperm motility. Methods Mice model of smoking was established, sperm samples were separated by Percoll discontinuous density gradients and sperm motility was detected, mice sperm CatSper mRNA was determined by RT-PCR, the distribution of CatSper protein was observed by immunocytochemical technique, CatSper protein level was detected by western blotting. Results In normal control group, mild smoking group, moderate smoking group, and severe smoking group, the proportion of sperm motility （a ＋ b） grade were （87.6 ± 16.2）% vs. （72.9 ± 13.0）% vs. （68.1± 16.0）% vs. （44.6± 15.0）% （P 〈 0.05）, the relative contents of CatSper mRNA were 0.915 ±0.202 vs. 0.712 ±0.121 vs. 0.522 ± 0.126 vs. 0.258 ± 0.177 （P 〈 0.05）, the relative contents of CatSper protein were 0.882 ± 0.208 vs. 0.693 ± 0.202 vs. 0.471 ±0.251 vs. 0.263 ±0.198 （P 〈 0.05）. CatSper protein expression was depressed by smoking in testis testified by immunocytochemical technique, and the results indicated significant negative correlation between CatSper mRNA/ protein and the proportion of sperm motility （a ＋ b） grade. Conclusions The expression of cation channel CatSper was significant inhibited by smoking with a positive correlation with sperm motility, which suggest that the lower expression of CatSper may be involved in the molecular mechanism of male reproductions impairment by smoking.

关 键 词：吸烟 钙离子通道蛋白CatSper 精子活力 

分 类 号：R163.2[医药卫生—公共卫生与预防医学]