Wuzi Yanzong Wan Regulates The Permeability Of Tight Junctions Of Sertoli Cells In Mice

Recently,male infertility has becomed a hot and difficult topic in the field of reproductive medicine in the world.Spermatogenic dysfunction is a major cause of male infertility.According to the traditional Chinese medicine,the kidney stores essence and governs reproduction,and kidney deficiency can result in spermatogenic dysfunction.Wuzi Yanzong Pills is a classic formula in the clinical treatment of male infertility,so its mechanism of improving spermatogenic function has attracted people's attention widely.The timely restructuring of testis Sertoli cell tight junction is known to play an important role in spermatogenesis.Whether Wuzi Yanzong Pills can regulate tight junctions remains to be further studied.Regarding the Sertoli cell tight junction permeability as the breakthrough point,this research explores the mechanism of the spermatogenic function of Wuzi Yanzong Pills in vivo experiments.The experimental research is mainly divided into the following four parts.Objective:To investigate the effect of Wuzi Yanzong Pills on Sertoli cells tight junction permeability.Methods:Fifty male C57 mice were randomly divided into five groups,i.e.the control group,the model group,the low-dose Wuzi Yanzong Pills(WYP)group,the medium-dose WYP group and the high-dose WYP group,respectively,10 in each group.The animals were treated daily with vehicle,glycosides tripterygium wilfordil(GTW)either alone(30mg/kg·d)or followed by WYP(1.5,3,or 6.0g/kg·d),respectively for 40 days.Biotin tracing method was used to evaluate the permeability of Sertoli cell tight junction.The levels of proinflammatory cytokines including IL-la?IL-6?IL-17a and TNF-a were analyzed by ELISA.The expression levels of tight junction-associated proteins including ZO-1?JAM-A?Occludin were analyzed by Western Blot.The ultrastructure of tight junction was observed by the transmission electron microscope.Results:Mice in the model group exhibited increased tight junction permeability and increased IL-la,IL-6,IL-17a and TNF-a levels compared with the control group.Whereas there was no significant change in the expression levels of ZO-1?JAM-A and Occludin.At the same time,the model group showed a ultrastructural damage in the testis,which showed vacuolization in Sertoli cells and abnormal structures in tight junction.Compared with the model group,the permeability of tight junction showed normal and the levels of IL-1?,IL-6,IL-17a and TNF-a were decreased.At the same time,the ultrastructural of the testis was complete in the WYP groups.There was no significant change in the expression levels of ZO-1?JAM-A and Occludin.Conclusion:Wuzi Yanzong Pill can improve spermatogenic function by regulating the testicular Sertoli cells tight junction structure and function.