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Objective To explore the pharmacological mechanism of Wuzi Yanzong Pill in the treatment of male spermatogenic dysfunction by network pharmacology in rat sertoli cells. Methods Targets of anti-spermatogenic dysfunction in Wuzi Yanzong Pill were screened and the biological function and signal pathway were analyzed by online database. The heat stress model was constructed with rat sertoli cells to detect changes in the production of reactive oxygen species (ROS) and the expression of Hsp70. Results The results of network pharmacology showed that Wuzi Yanzong Pill mainly improved male spermatogenesis dysfunction by regulating ROS level and extracellular stimulus response etc. The ROS fluorescence probe results showed that，compared with the blank group，the ROS production was significantly increased in the model group，and significantly decreased in the Wuzi Yanzong Pill group. Western blot results showed that the expression of Hsp70 in Wuzi Yanzong Pill group was higher than that in the blank group and lower than that in the model group. Conclusion Wuzi Yanzong Pill seems to protect rat sertoli cells under heat stress by reducing ROS production and Hsp70 expression. [ABSTRACT FROM AUTHOR]

Objective To discuss the effect and mechanism of Wuzi Yanzong Wan (Five Seeds for Multiplying Pills) in the proliferation of Sertoli cells in infant rats. Methods The Sertoli cells were separated from male SD rats (20 d old) and cultured, and then randomly divided into normal group, low-dose Wuzi Yanzong Wan group (5 g/L, low-dose group), mid-dose Wuzi Yanzong Wan group (10 g/L, mid-dose group) and high-dose Wuzi Yanzong Wan group (15 g/L, high-dose group). The changes of optical density (OD) of Sertoli cells were detected by using methyl thiazolyl tetrazolium test (MTT) in all groups. The expression of Ki67 positive granules in Sertoli cells were detected by using immunohistochemistry technique in all groups. The expressions of Akt and p-Akt in Sertoli cells were detected by using Western blot method. Results Compared with normal group, the proliferation of Sertoli cells treated with Wuzi Yanzong Wan was promoted significantly (P<0.05 or P<0.01). The expression of Ki67 positive granules increased significantly (P<0.05 or P<0.01), and relative expression of p-Akt increased significantly (P<0.05 or P<0.01). Conclusion Wuzi Yanzong Wan can possibly promote the proliferation of Sertoli cells in infant rats through activating Akt signal transduction pathway and up-regulating p-Akt expression. [ABSTRACT FROM AUTHOR]

Objective To study the mechanism of Wuzi Yanzong Wan (Five-Seed Progeny Pill) on im­proving heat stress resistance of Sertoli cells in rats based on data from the whole genome microarray. Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan (WZYZW) group, normal group and model group. The cells in three groups were given WZYZW solution and equal volume of PBS respectively. After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 °C for 15 minutes, while rats in normal group were put in water bath at 37 °C for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technolo­gy. Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated. There were totally 473 differently expressed genes in the normal group, including 168 up-regu-later! genes and 305 down regulated genes. Among these down-regulated gene expressions. RGD1305298. Trptl, D113, Abi3 and Ppplrl2b were significant in the WZYZW group and norma! group, while Dusp7 was significant among up-regulated gene expressions. The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regula­tion of act in cytoskeleton, MAPK signaling pathway and tight junction pathway, etc. Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of Sertoli cells may involve mainte­nance of tight junctions between Sertoli cells, inhibition of Sertoli cells apoptosis and promotion of cell dif­ferentiation and maturation. [ABSTRACT FROM AUTHOR]