Chaize mixture alleviates NAFLD by regulating Glipr2-related autophagy/NLRP3 signaling

Chaize Mixture (CZM), formulated by combining Xiaochaihu Decoction and Zexie Decoction, is a time-tested Chinese herbal prescription historically utilized for managing metabolic disorders, including non-alcoholic fatty liver disease (NAFLD). Despite its extensive historical use, the precise mechanisms underlying its therapeutic efficacy remain to be fully elucidated. This study aimed to delineate the molecular mechanisms of CZM in both in vivo and in vitro models of NAFLD. It was found that CZM treatment led to decreased serum ALT (alanine aminotransferase) and AST (aspartate aminotransferase), hepatic TG (triglycerides), and reduced of Srebp1, Fas, Tnf-α, IL-6, IL-1β, Tgf-β, Timp1, Timp2, Acta2, and Adamstl2, while elevating Cpt1a levels. Histopathological assessment revealed that CZM alleviated inflammatory cell infiltration, and fat accumulation in hepatocyte. CZM could increase gut microbial diversity and richness. Further multi-technique comprehensive analysis showed that Glipr2 expression was elevated in the model group but significantly decreased after CZM, which was verified by PCR and Western blotting. In addition, CZM decreased microtubule-associated protein 1 light chain 3B (LC3B), p62, NLRP3, IL-1β, Caspase1 protein levels, and increased the levels of Beclin1. CONCLUSION: Our results showed that CZM could ameliorate steatosis, inflammatory response, and fibrosis in NAFLD by promoting autophagy and inhibiting NLRP3-mediated inflammation, which was achieved through regulating Glipr2.