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目的:探讨附睾蛋白酶抑制剂(epididymal protease inhibitor,EPPIN)基因在男性不育患者精子中的表达情况及其对男性生育力的影响。方法:收集少弱精子症组(n=8)、弱精子症组(n=12)及对照组(n=19)的精液标本,分别采用Western blotting、半定量RT-PCR和实时荧光定量PCR(qRT-PCR)方法检测各组精子中EPPIN的表达,并计算其相对表达量及与精液质量的相关性。结果:EPPIN基因在不同组精子中均有表达,在对照组、弱精子症组和少弱精子症组的EPPIN mRNA的相对表达量分别为0.09±0.02、0.29±0.08和1.00±0.18,各组间比较差异有显著统计学意义(P<0.01)。EPPIN蛋白的相对表达量与mRNA类似。EPPIN mRNA表达量与精子浓度、前向运动精子(progressive motility,PR)百分率和活力呈显著负相关。结论:EPPIN可能与人类精子数量及活力有关,推测可能参与少弱精子症的发病机制。
Objective: To investigate the expression of epididymal protease inhibitor (EPPIN) gene in sperm of male infertility patients and its effect on male fertility. Methods: Semen samples of oligoasthenospermia group (n = 8), asthenospermia group (n = 12) and control group (n = 19) were collected and analyzed by Western blotting, semi-quantitative RT-PCR and real- (qRT-PCR) method was used to detect the expression of EPPIN in each group of spermatozoa. The relative expression of EPPIN and sperm quality were calculated. Results: The expressions of EPPIN mRNA in different groups of sperms were 0.99 ± 0.02,0.29 ± 0.08 and 1.00 ± 0.18 in control group, asthenospermia group and azoospermia group, respectively The difference was statistically significant (P <0.01). The relative expression level of EPPIN protein is similar to that of mRNA. EPPIN mRNA expression was negatively correlated with sperm concentration, percentage of progressive motility (PR) and vitality. Conclusion: EPPIN may be related to human sperm quantity and vitality, suggesting that it may be involved in the pathogenesis of oligoasthenospermia.