柯萨奇病毒-腺病毒受体siRNA对体外血睾屏障完整性的影响

Effects of coxsackie-adenovirus receptor-specific small interfering RNA duplexes on the integrity of in vitro blood-testis barrier

目的观察柯萨奇病毒-腺病毒受体(CAR)siRNA对体外血睾屏障完整性的影响,探讨睾丸CAR分子功能。方法体外建立血睾屏障模型,以CAR siRNA和阴性对照siRNA进行转染,然后以RTPCR和Western blot法检测相关基因mRNA或蛋白表达的变化,免疫荧光法检测蛋白分布,以及测量跨上皮电阻的改变。结果 CAR siRNA组较对照组CAR表达显著降低。与对照组比较,CAR siRNA显著降低了跨上皮电阻,同时,也显著降低了ZO-1的表达,使其分布发生紊乱。结论 CAR siRNA降低支持细胞上皮屏障功能,CAR是血睾屏障的重要调控分子。

Objective To investigate the function of coxsackie-adenovirus receptor（ CAR） in testis by observing the effects of CAR-specific small interfering（ si） RNA duplexes on the integrity of in vitro blood-testis barrier. Methods The in vitro Sertoli cell blood-testis barrier was established. Cells were transfected with nontargeting or CAR-specific siRNA duplexes. The mRNA and protein levels or the intracellular distribution of related genes were measured. The transepithelial electrical resistance（ TER） was assessed. Results CAR-specific siRNA duplexes induced CAR knockdown in Sertoli cells,and significantly decreased TER,and led to a significant decrease in the protein expression of ZO-1. The distribution of ZO-1 was also interfered. Conclusion CAR-specific siRNA duplexes reduces the function of Sertoli cell bloodtestis barrier,indicating CAR is an important molecule involved in regulating the blood-testis barrier.