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Wu Zi Yan Zong Wan Improves Triptolide-Induced Testicular Spermatogenic Dysfunction Through the Spermatogenic
Lei Zhang1,2, Binghao Bao2, Weizhen Wu2,3
1Department of Traditional Chinese Medicine, The First People's Hospital of Lianyungang, Jiangsu, China.
None:
Wu Zi Yan Zong Wan (WZYZW) is a traditional Chinese formula known for treating male infertility, though its mechanism of action is unclear. This study investigates how WZYZW may alleviate spermatogenic dysfunction in mouse testes induced by triptolide (TP). We established a TP-induced spermatogenic dysfunction mouse model and randomly divided 30 male C57BL/6J mice into five groups: control, model, and low-, medium-, and high-dose WZYZW groups (six mice per group). After a 35-day intervention, testicular tissues were analyzed for histopathology via hematoxylin-eosin and TUNEL staining. Protein expression of type I collagen, laminin, fibronectin, and spermatogonial stem cell (SSC) markers, including GFRα1, Ret, and PLZF, were evaluated by immunohistochemical staining. GDNF and CSF1 levels were measured by ELISA, and the PLCγ/PKC pathway was analyzed via Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Compared with the model group, WZYZW significantly increased testicular index (high-dose, p < .01), improved Johnsen score (all doses, p < .05), reduced spermatogenic cell apoptosis, and improved the spermatogenic microenvironment by increasings type I collagen (high-dose, p < .01), laminin (low-dose, p < .01; medium/high-dose, p < .05), fibronectin (high-dose, p < .01), GDNF (medium/high-dose, p < .05), and CSF1 (high-dose, p < .05). It significantly upregulated the expression of GFRα1, PLZF, C-kit, Ret, and SCF, with the improvement of GFRα1(medium/high-dose vs. low-dose, p < .01), Ret (medium/high-dose vs. low-dose, p < .05), SCF (high-dose vs. low-dose, p < .05) showing a certain dose-dependency. The qRT-PCR and Western blotting results showed that WZYZW significantly enhanced PLCγ/PKC pathway activity. The study indicates that WZYZW can improve the spermatogenic microenvironment and promote the self-renewal and differentiation of SSCs through the PLCγ/PKC pathway, thereby improving spermatogenic function.
