无精子症和生精功能正常者睾丸组织中血红素加氧酶的表达与差异性研究  

Expression of heme oxygenase enzyme in the testis tissue and azoospermia

作　　者：池艳[1] 毛向明[2] 张欣宗[1] 张峰彬[3] 顾宇春[4] 熊承良[1] 

机构地区：[1]华中科技大学同济医学院计划生育研究所/生殖医学中心专科医院,湖北武汉430030 [2]南方医科大学南方医院泌尿外科,广东广州510515 [3]浙江大学附属妇产科医院,浙江杭州310023 [4]英国伯明翰大学医学院

出　　处：《中华男科学杂志》2011年第8期712-716,共5页National Journal of Andrology

摘　　要：目的:对人类睾丸组织中表达血红素加氧酶(HO)的细胞进行定位;通过测定原发性无精子症及梗阻性无精子症患者睾丸组织中血红素加氧酶1(HO-1)的表达量与正常睾丸组织中HO-1表达量的差异性,来探讨其与无精子症发病的相关性。方法:应用免疫组化方法对人类睾丸组织中表达HO的细胞进行定位;采用逆转录-荧光定量PCR(FQ-PCR)方法定量检测无精子症患者与正常人睾丸组织HO-1及HO-2基因水平的表达量;应用W est-ern印迹检测各组之间HO蛋白水平表达量。结果:在正常睾丸组织,HO-1主要表达在支持细胞上;而HO-2在支持细胞和各级生精细胞中均有表达;FQ-PCR结果显示非梗阻性无精子症患者睾丸组织HO-1、HO-2的表达量均显著低于正常组及梗阻性无精子症组(P<0.05),差异具有统计学意义。而梗阻性无精子症患者睾丸组织表达HO-1、HO-2的量与正常组相比无显著性差异。W estern印迹结果显示HO-1蛋白水平的表达量差异与基因水平一致。而HO-2的蛋白水平在各组之间表达没有显著性差异。结论:非梗阻性无精子症患者睾丸组织中HO的表达量显著性降低,且HO-1无论是蛋白水平还是基因水平的差异一致。HO-1可以通过抗炎、抗氧化、抗凋亡的机制保护睾丸组织免受各种应激的损伤,从而维护正常的生精功能。可见,HO-1的减少可能与生精功能低下相关,这可能是非梗阻性无精子症的发病机制之一。Objective： To investigate the location of heme oxygenase （HO） enzyme in the human testis, and explore the correlation of the expression of HO enzyme with azoospermia by analyzing its different expression levels in the testes of nonobstructive azoospermia, obstructive azoospermia and normal men. Methods ： We detected the location of the cells expressing HO enzyme in the human testis tissue using immunohistochemistry, determined the mRNA and protein expression levels of HO-1 and HO-2 in the testes of azoospermia patients and normal healthy men by RT-tluorescence quantitative PCR （RT-FQ-PCR） and Western blot, and explored the correlation of HO expressions with the pathogenesis of azoospermia. Results ： HO-1 enzyme was expressed mainly in the Sertoli cells and HO-2 enzyme chiefly in the germ cells of the testis tissue. RT-FQ-PCR showed that the expression of HO-1 in the testis tissue was significantly lower in the nonobstructive azoospermia than in the normal and obstructive azoospermia groups （P 〈 0.05 ）, with no significant difference between the latter two. Western blot revealed no obvious difference between the expression level of HO-1 protein and that of HO-1 mRNA. There were no differences in the expression level of HO-2 protein among the three groups. Conclusion ： The expression level of HO enzyme is significantly decreased in the testis tissue of nonobstruetive azoospermia patients, and the expression of HO-1 protein is consistent with that of HO-1 mRNA. As HO-1 protects the testis tissue against various stress injuries through its antioxidant, anti-inflammatory and anti-apoptotie effects, its decreased expression level may he correlated with spermatogenie dysfunction, and therefore considered as a possible mechanism of nonobstrnetive azoospermia.

关 键 词：血红素加氧酶 无精子症 睾丸 

分 类 号：R698.2[医药卫生—泌尿科学]