人精子膜蛋白P34H表达与透明质酸酶活性的关系

Correlation between the expression of P34H and the activity of hyaluronidase in human spermatozoa

目的探讨人精子膜蛋白P34H表达和顶体内透明质酸酶活性的关系。方法收集88例精液标本,其中正常生育组20例,不育男性68例。参照世界卫生组织(WHO)标准对标本进行精液常规分析,根据精液参数的不同,将68例不育标本分为正常精液参数不育组和精液参数异常不育组。Western blotting检测P34H蛋白在人精子中的表达,免疫荧光观察P34H蛋白在精子表面的阳性表达率,采用改良透明质酸钠-明胶底物膜法检测精子顶体内透明质酸酶(HYD)活性(HYD阳性反应率、HYD活性强度)。结果正常精液参数不育组和精液参数异常不育组的精子P34H/β-actin平均吸光度、精子P34H标记阳性率均明显低于正常生育组,经统计学分析差异均有显著性(P<0.05);正常精液参数不育组和精液参数异常不育组的HYD活性(HYD阳性反应率、HYD活性强度)与正常生育组比较均有显著下降(P<0.05)。相关性分析提示,精子P34H蛋白表达量与HYD活性(HYD阳性反应率、HYD活性强度)存在正相关性,相关系数r=0.449、0.431,P<0.01;精子P34H阳性率与HYD活性(HYD阳性反应率、HYD活性强度)存在正相关性,相关系数r=0.727、0.691,P<0.01。结论男性不育患者精子P34H蛋白表达减少,精子P34H阳性率降低以及HYD活性(HYD阳性反应率、HYD活性强度)减弱。

Objective To investigate the correlation between the level of P34 H expression and the activity of hyaluronidase in human spermatozoa. Methods Eighty eight semen samples were collected,68 cases were in the infertile group,and 20 in the normal control group. Semen routine analysis was referred to the WHO standard method. According to the difference of semen parameters,68 cases of infertile males were divided into the infertile group with normal semen parameters and the abnormal semen parameters. Western blotting was used to detect the level of P34 H expression on spermatozoa. The P34H-positive rate on human spermatozoa was determined by indirect immunofluorescent staining using anti-P34 H antibody. The HYD-positive rate and HYD-activity intensity in all samples were examined by improved fixedsubstrate film method. Results The level of P34 H protein expression and the percentage of the P34H-positive rate in infertile groups with normal semen parameters and abnormal semen parameters were significantly lower than that in the control group（ P〈0.05）. The activity of HYD（ HYD-positive rate,HYD-activity intensity） in the infertile groups with normal semen parameters and abnormal semen parameters were also significantly lower than that in control group（ P〈0.05）. The relation between the P34 H protein expression and HYD-positive rate,HYD-activity intensity had a significant positive correlation（ r = 0. 449,0. 431; P〈0.01）; the relation between the the percentage of P34H-positive rate and HYDpositive rate,HYD-activity intensity had a significant positive correlation（ r = 0. 727,0. 691; P〈0.01）. ConclusionThe level of P34 H protein expression and the percentage of the P34H-positive rate are decreased,while the activity of HYD（ HYD-positive rate,HYD-activity intensity） is reduced in male infertility.