ETHNOPHARMACOLOGICAL RELEVANCE:Chaihu Guizhi Ganjiang Decoction (CGGD), a classical traditional Chinese medicine (TCM), has demonstrated efficacy against pancreatic fibrosis of chronic pancreatitis (CP). However, its regulatory effects on other key pathological aspects of CP remain unclear.
AIM OF THE STUDY:This study aimed to investigate the protective effects of CGGD against pancreatic acinar cell ferroptosis of CP and elucidated the underlying mechanisms.
MATERIALS AND METHODS:The CP rat model was induced by dibutyltin dichloride (DBTC) for 4 weeks, and the efficacy of CGGD on pancreatic injury and fibrosis was evaluated based on serum analysis and histopathological examination. Network pharmacology combined with proteomics analysis identified potential therapeutic targets of CGGD for CP. Transmission electron microscopy (TEM) confirmed characteristic morphological hallmarks of ferroptosis. Biochemical assays were used to measure tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), as well as iron content. The key mediators of ferroptosis (TFR1, FTH1, SLC7A11, GPX4, p53) were examined by RT-qPCR and Western blot, and GPX4 and p53 localization was further confirmed by immunofluorescence. In vitro, AR42J cells were treated with cerulein, CGGD drug-containing serum (CGGDs), Nutlin-3a (a p53 activator), Erastin (a SLC7A11 inhibitor), or Trp53 siRNA. The key mediators of inflammation and ferroptosis were evaluated using commercial kits, RT-qPCR, Western blot, and immunofluorescence staining.
RESULTS:CGGD significantly reduced pancreatic injury, cell death, and fibrosis in DBTC-induced CP rats. Mechanistically, p53 and ferroptosis were identified as critical targets of CGGD in CP. Specifically, CGGD suppressed p53 phosphorylation, inhibited ferroptosis pathway activation, and diminished oxidative stress both in vivo and in vitro. Notably, the reductions of ROS and restoration of GSH induced by CGGD were markedly reversed by cotreatment with Nutlin-3a or Erastin in cerulein-induced AR42J cells. Furthermore, the effectiveness of CGGDs in mitigating ferroptosis and oxidative stress was not further weakened following the knockdown of Trp53.
CONCLUSION:Our findings demonstrate that CGGD suppresses ferroptosis in pancreatic acinar cells via the p53/SLC7A11/GPX4 axis, thereby elucidating a novel therapeutic mechanism of CGGD for the treatment of CP.