糖酵解途径在人精子顶体反应的作用研究  

The role of glycolytic pathway in acrosome reaction in human spermatozoa

作　　者：张继良[1] 王朋[1] 戴数[1] 祝文婷[2] 张皓[2] 

机构地区：[1]丽水市人民医院检验科,浙江丽水323000 [2]上海交通大学医学院附属新华医院教育部和上海市环境与儿童健康重点实验室,上海200092

出　　处：《中国卫生检验杂志》2016年第15期2168-2170,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金资助项目(81401253)

摘　　要：目的探讨糖酵解途径在人精子顶体反应的作用及其分子机制。方法取健康志愿者捐献的精液,采用体外实验的方法,以糖酵解抑制剂S-3-氯-1,2-丙二醇(S-3-MCPD)处理人精子,处理组浓度分别为0.1 mmol/L、0.5 mmol/L和2.5 mmol/L,对照组加入相同体积溶剂。用考染法检测顶体反应发生率,免疫印迹法检测蛋白酪氨酸磷酸化,虫荧光素酶法检测ATP水平。结果 0.5 mmol/L和2.5 mmol/L S-3-MCPD处理组顶体反应发生率分别为34.5%和25.9%,与对照组(57.2%)比较,差异有统计学意义(P<0.01)。S-3-MCPD蛋白酪氨酸磷酸化存在显著剂量-效应关系。ATP含量与S-3-MCPD作用浓度呈显著负相关(r=0.769,P<0.01)。结论糖酵解途径在人精子顶体反应发生过程具有重要作用,其机制可能涉及糖酵解产生的ATP为蛋白酪氨酸磷酸化所需,而后者则是顶体反应的关键调节机制之一。Objective To investigate the role of glycolysis pathway in acrosome reaction in human spermatozoa and to explore the potential molecular mechanism. Methods The human sperm isolated from healthy volunteers donated semen were treated with S- 3- Chloro- 1,2- propanediol（ S- 3- MCPD）,a sperm- specific glycolytic inhibitor,with the final concentration of0. 1 mmol / L,0. 5 mmol / L and 2. 5 mmol / L,respectively. The same volume of vehicle was added into the control. Acrosome reaction was assessed with Coomassie stain,and protein tyrosine phosphorylation was measured with immunoblotting,and ATP level was measured using luciferase kit. Results The acrosome reaction rate of human sperm treated with 0. 5 mmol / L and2. 5 mmol / L S- 3- MCPD were 34. 5% and 25. 9%,respectively,and significantly lower than that in the control（ 57. 2%）,with the difference statistically significant（ P〈0. 01）. There was significant dose- effect relationship between S- 3- MCPD and tyrosine phosphorylation. The ATP level also showed a negative correlation with the effective concentration of S- 3- MCPD（ r =0. 769,P〈0. 01）. Conclusion Glycolysis plays a role in acrosome reaction in human sperm,and the mechanism may be involved in that the ATP generated from glycolysis are required for protein tyrosine phosphorylation,which is the key regulation of acrosome reaction.

关 键 词：糖酵解途径 顶体反应 蛋白酪氨酸磷酸化 ATP 人精子 

分 类 号：R446.19[医药卫生—诊断学]