弱精子症患者精子外周致密纤维2基因及蛋白的表达  

Expressions of ODF2 mRNA and protein are down-regulated in the sperm of asthenospermia patients

作　　者：罗斌[1] 何维[2] 王世恒[2] 熊洪平 梁鑫[1] 单旭东[1] 吕自力[1] 

机构地区：[1]成都中医药大学第二附属医院生殖医学中心,四川成都610041 [2]成都中医药大学第二附属医院检验科,四川成都610041

出　　处：《中华男科学杂志》2017年第11期1002-1006,共5页National Journal of Andrology

基　　金：四川省科技厅科研院所成果转化项目(15010116);成都市科技项目(2014-HM01-00187-SF);成都中医药大学校基金(ZRQN1402);成都中医药大学科技发展基金(ZRYY1635)~~

摘　　要：目的:探讨弱精子症患者精子中外周致密纤维2(ODF2)mRNA及蛋白的表达水平,比较ODF2在健康男性与弱精子症患者中的表达差异。方法:根据WHO第5版,收集身体健康无疾病正常男性精液(n=15)和弱精子症患者精液(n=45)共60份,利用计算机辅助精液分析系统(CASA)进行精子统计,将弱精子症分为轻、中、重度3组,采用实时定量PCR和Western印迹对4组标本进行ODF2基因的表达量研究。结果:与正常组相比,轻度弱精子症组精子中ODF2基因表达无明显统计学差异(1.1120±0.5255 vs 0.6880±0.3720,P>0.05);中度弱精子症组ODF2基因表达降低,有显著的统计学差异(1.1120±0.5255 vs 0.4833±0.1863,P<0.05);重度弱精子症组ODF2基因表达明显降低,有极显著的统计学差异(1.1120±0.5255 vs 0.448 3±0.3408,P<0.01)。Western印迹中ODF2的OD值(ODF2/β-actin)在正常组和轻、中、重度弱精子症组分别为:0.4587±0.0521、0.3261±0.0714、0.1454±0.0536和0.1227±0.0457,与正常组相比,轻度弱精子症组无明显统计学差异(P>0.05),中度弱精子症组有显著的统计学差异(P<0.05),重度弱精子症组有极显著的统计学差异(P<0.01)。结论:ODF2基因及蛋白在弱精子症患者精子中的表达下调与精子活动力下降有密切联系,呈正相关性,提示ODF2基因可能参与精子活力的调控。Objective ： To investigate the mRNA and protein expressions of outer dense fiber 2 （ ODF2 ） in the sperm of the as- thenospermia patient and their differences from those in normal healthy men. Methods： According to the WHO criteria, we collected semen samples from 45 asthenozoospermia patients and 15 normal healthy volunteers. Using computer-assisted sperm analysis （ CA- SA） , we divided the semen samples from the asthenospermia patients into a mild, a moderate and a severe group, and determined tile mRNA and protein expressions of ODF2 in different groups by RT-PCR and Western blot. Results ： Compared with the normal healthy men, the expression of the ODF2 gene showed no statistically significant difference in the mild asthenospermia group （ 1. 112 0 ± 0. 525 5 vs O. 688 0 ± 0. 372 0, P 〉 0.05） but remarkably decreased in the moderate （0. 483 3 ± 0. 186 3, P 〈 0.05） arid severe asthenospermia patients （0. 448 3±0. 340 8, P 〈 0.01 ）. The OD value （ODF2/13-actin） of the ODF2 protein in the normal men exhibited no statistically significant difference from that in the mild asthenospermia group （0. 458 7 ±0. 052 1 vs O. 326 1 ± 0. 071 4, P 〉 0.05） , but markedly lower than in the moderate （0. 145 4 ± 0. 053 6, P 〈 0.05） and severe asthenospermia patients （0. 122 7 ± 0. 045 7, P 〈 0.01 ）, which was consistent with the results of RT-PCR. Conclusion： Decreased mRNA and protein expressions of ODF2 in the sperm are positively correlated with declined sperm motility of the asthenospermia patient, which is suggestive of the in-volvement of the ODF2 gene in the regulation of sperm motility

关 键 词：外周致密纤维2 弱精子症 实时定量PCR 蛋白印迹 

分 类 号：R698.2[医药卫生—泌尿科学]