Ethnopharmacological relevance: Wu-Zi-Yan-Zong-Wan (WZYZW) is a classical traditonal Chinese herbal formula and a Chinese patent medicine used to treat male infertility. However, the chemical components of WZYZW and its mechanism are not yet fully clarified. Aim of the study: The purpose of this study is to observe the effect and underlying mechanism of WZYZW on ameliorating blood-testis barrier (BTB) dysfunction in mice with spermatogenic dysfunction induced by administration of Tripterygium wilfordii Hook. f. multiglycosides (GTW). Materials and methods: WZYZW was administered by gavage to mice with GTW-induced spermatogenic dysfunction (kidney essence deficiency pattern) for 40 days. Testis tissues were obtained for subsequent histopathological analysis. Biotin tracing was used to evaluate the permeability of Sertoli cell tight junctions. The levels of proinflammatory cytokines including interleukin (IL)-6, IL-17A, IL-1 alpha and tumor necrosis factor (TNF)-alpha were analyzed by ELISA. The expression levels of proteins related to tight junction including ZO-1, JAM-A and occludin were analyzed by western blotting. The ultrastructures of tight junctions were observed by transmission electron microscopy. Results: WZYZW ameliorated GTW-induced testicular spermatogenic dysfunction. Levels of IL-6, IL-17A, IL-1 alpha, and TNF-alpha in the groups receiving low, medium, and high doses of WZYZW decreased in a dose-dependent manner. WZYZW impeded a biotin tracer from permeating the BTB, protecting its integrity in GTW-treated mice. In addition, our results showed no significant changes in the protein expressions of ZO-1, JAM-A, and occludin after WZYZW administration compared with the GTW group. Meanwhile, WZYZW exhibited a linear arrangement and restored the typical "sandwich" structure of BTB. No acute poisoning incidences were observed in all groups during the experiment. Conclusions: Our findings demonstrate that WZYZW may ameliorate some GTW-induced BTB dysfunction, possibly by regulating proinflammatory cytokine levels. In vitro studies on the regulation of BTB permeability by WZYZW and its active components are further required.