弱精子症动物模型中尿纤溶酶原激活因子的含量及表达情况研究

Correlation of the Content and Expression of Urokinase Plasminogen Activator with Asthenospermia in Rat Models

目的:利用奥硝唑所致弱精子症动物模型,采用免疫组化方法和RT-PCR技术,了解尿纤溶酶原激活因子(uPA)在弱精子症动物模型中的含量及表达情况,初步探讨奥硝唑所致弱精子症动物模型的机制及uPA作为预防或治疗弱精子症药物的可能性。方法:48只雄性SD大鼠随机分为1d用药组,5d用药组,10d用药组,15d用药组,20d用药组和对照组,每组8只,用药组每天给予奥硝唑200mg/kg,对照组给予0.5%羧甲基维素钠连续灌胃,用药组分别在给药第1、5、10、15、20d后24h内,麻醉处死动物取附睾和睾丸。低渗肿胀试验检测精子细胞膜完整性,免疫组化方法动态观察睾丸与附睾组织中uPA蛋白表达情况,RT-PCR检测睾丸组织中uPAmRNA含量。结果:奥硝唑持续给药构建弱精子症时,精子膜完整性下降发生在给药的第10d,并一直维持低值。与建立弱精子症模型同步的uPA在睾丸和附睾组织蛋白表达及mRNA含量下降在用药15d,下降趋势上是平行的,而显效性稍滞后。用药15d组与用药20d组uPA蛋白表达、mRNA水平分别与对照组比较有统计学意义(P<0.05)。结论:精子细胞膜受损、运动能力下降与uPA表达及含量下降平行,奥硝唑所致弱精子症模型形成可能是由于uPA含量及表达下降所致。弱精子症形成原因较复杂,uPA含量及表达的下降可认为是弱精子症形成因素之一,检测uPA含量可能有助于弱精子症的诊断和预防。

Objective： To investigate the content and expression of the urokinase plasminogen activator （uPA） in the ornidazole-induced astbenospermia animal model, and to probe the mechanism of ornidazole inducing astbenospermia and the possibility of using UPA for the prevention and treatment of asthenospermia. Methods ： Forty-eight male rats were equally randomized into 5 medication groups （1 d, 5 d, 10 d, 15 d and 20 d） and a blank control group, and ornidazole （200 mg/kg） was given intragastrically every day to the former five while 0.5% carboxymethylcellulose Na （CMC-Na） to the latter for 20 successive days. Then the rats were sacrificed by intraperitoneal injection of pentobarbital at 1, 5, 10, 15 and 20 days respectively and the epididymides and testes harvested. The integrity of the sperm cell membrane was detected by hypoosmotic swelling experiments, the UPA expression in the testicular and epididymal tissues dynamically observed by immunohistochemistry and the level of uPA mRNA in the testis determined by RT-PCR. Resuits： The integrity of the sperm cell membrane was reduced at 10 days and remained low till the end of the medication, but with no statistic significance. Compared with the blank controls, the uPA expression and mRNA content in the testicular and epididymal tissues showed no conspicuous difference in the 1 d and 5 d groups, decreased insignificantly in the 10 d group, but significantly in the 15 d and 20 d groups （ P 〈 0.05 ）. Conclusion ： The defect of sperm cell membrane and decrease of sperm motility go in parallel with the reduced expression and content of uPA, which may be one of the factors for the development of asthenospermia.