Zuogui Pill alleviates chemotherapy-induced ovarian aging and oogonial stem cell injury by modulating cGAS-STING-mediated inflammatory signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Zuogui Pill (ZGP), a representative prescription for tonifying kidney and replenishing essence, is commonly employed in treating ovarian aging and related reproductive disorders. AIM OF THE STUDY: To investigate the effects of ZGP on chemotherapy-induced ovarian aging and oogonial stem cell (OSC) injury, with an emphasis on the involvement of cGAS-STING signaling. MATERIALS AND METHODS: UPLC-Q-TOF-MS was used to identify representative constituents of Zuogui Pill (ZGP). A chemotherapy-induced ovarian aging model was established by administering cyclophosphamide (CTX, 120 mg/kg) via intraperitoneal injection, and mice received oral ZGP treatment for six weeks. Phosphoramide mustard cyclohexanamine (PM)-exposed OSCs served as an in vitro model and were treated with ZGP-containing serum. The involvement of STING signaling was further examined through STING knockdown and pharmacological STING activation with DMXAA. Hematoxylin-eosin staining, ELISA, immunohistochemistry, western blot, qPCR, and immunofluorescence were performed to evaluate ovarian histomorphology, serum sex hormones, inflammatory responses, the expression of germline- and stem cell-associated markers, apoptosis-related changes, and pathway activation. RESULTS: ZGP improved serum sex hormone profiles, increased ovarian index, and promoted follicular development in CTX-treated mice. In both animal and cellular models, ZGP attenuated cGAS-STING pathway activation and reduced inflammatory cytokine and interferon-stimulated gene expression. ZGP also increased the expression of Oct4, MVH, and Fragilis, reduced the expression of P21, and ameliorated alterations in apoptosis-related proteins. STING knockdown alleviated PM-induced OSC injury, whereas ZGP attenuated DMXAA-stimulated STING activation. CONCLUSION: ZGP alleviated chemotherapy-induced ovarian aging and OSC injury, with improvements in ovarian morphology, sex hormone profiles, and germline- and stem cell-associated marker expression, as well as reduction in inflammatory and apoptosis-related changes. The effects of ZGP may involve modulation of cGAS-STING-mediated inflammatory signaling. These findings provide further insight into the potential clinical application of ZGP in chemotherapy-induced ovarian aging.