Xin Jia congrong tusizi decoction reverses ferroptosis in granulosa cells to rescue ovarian function decline via p53/Nrf2/SLC7A11/GPX4 signaling pathway

  • J Ethnopharmacol. 2026 Aug 10:367:121659. doi: 10.1016/j.jep.2026.121659.
Min Liu  1 ,  Yi Sun  2 ,  Xue Liu  3 ,  Shaobin Wei  4 ,  Jing Yang  5 ,  Yan Wang  6 ,  Keming Wu  7 ,  Yeke Wu  8
Affiliations
  • 1. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 2. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 3. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 4. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 5. School of Bioscience and Technology, Chengdu Medical College, Chengdu, 610500, China. Electronic address: [email protected].
  • 6. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 7. Department of Gynecology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
  • 8. Department of Stomatology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China. Electronic address: [email protected].
PMID: 42000005 DOI: 10.1016/j.jep.2026.121659
Abstract

Ethnopharmacological relevance: Ovarian function decline (OFD) represents a principal contributor to infertility among women of reproductive age, yet no definitive therapy has been established. It has been suggested that Ferroptosis exists in OFD. A traditional Chinese medicine formula called Xin Jia Congrong Tusizi Decoction (XJCTD) has shown potential in improving OFD. However, the mechanism remains unclear.

Aim of the study: This study aims to elucidate the protective effect of XJCTD on ovarian function in OFD and to clarify its underlying molecular mechanisms.

Materials and methods: Triptolide (TP)-induced rat and COV434 cell models were established to evaluate XJCTD efficacy. The role of the p53-Nrf2 axis was interrogated using p53 overexpression plasmids, Nrf2 siRNA, and inhibitors Pifithrin-α. Ovarian pathological changes were assessed using H&E staining. The major compounds of XJCTD were identified by UPLC-Q-TOF-MSE analysis and network pharmacology analysis. Direct interactions between core compounds and p53 were validated using molecular docking, surface plasmon resonance (SPR); and the interaction between XJCTD and TP53 was confirmed by cellular thermal shift assay (CETSA). Expression of p53 and Nrf2 was analyzed by Western blot (WB) or immunofluorescence, and their physical interaction validated by co-immunoprecipitation. We evaluated the impact of XJCTD on ferroptosis-related markers in both ovarian tissues and COV434 cells, including Fe2+, MDA, ROS and GSH. Ferroptosis-related proteins were further examined by WB and immunohistochemistry.

Results: XJCTD significantly attenuated TP-induced ovarian injury and Ferroptosis by suppressing Fe2+, lipid peroxidation, and ROS accumulation while restoring GSH levels. Furthermore, XJCTD facilitated p53 ubiquitination and degradation, consequently alleviating Nrf2 suppression to activate the SLC7A11/GPX4 pathway. Rescue experiments demonstrated that p53 overexpression or Nrf2 silencing reversed these protective benefits. SPR analysis confirmed direct binding between p53 and core XJCTD bioactive components, including Epimedin A, Isoquercitrin, and Isoacteoside.

Conclusions: XJCTD functions as a suppressor of Ferroptosis to protect ovarian function. The underlying mechanism involves the targeted degradation of p53, which alleviates the inhibition of Nrf2 nuclear accumulation and enhances the SLC7A11/GPX4 antioxidant defense system.

Keywords
Ferroptosis; Ovarian function decline; Reactive oxygen species; Reproductive system disorders; Traditional Chinese medicine; p53/Nrf2/SLC7A11/GPX4 pathway.
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