Qianggan Ruanjian Pill ameliorates liver fibrosis through regulation of the TGF-β1/Smad and PI3K/AKT signalling pathways

Abstract

Ethnopharmacological relevanceLiver fibrosis is a critical pathological process in the progression of chronic liver injury, ultimately resulting in cirrhosis, for which currently available therapeutic interventions remain inadequate. Among these, the Qianggan Ruanjian Pill (QGRJP) has emerged as a clinically experienced formula with notable therapeutic efficacy against liver fibrosis. However, the precise underlying mechanisms require further investigation. Aim of the studyIn this study, we investigated the key pathways and target genes of QGRJP that attenuate liver fibrosis and elucidated the underlying mechanisms. Materials and methodsHigh-performance liquid chromatography-mass spectrometry (HPLC-MS) was used to identify the major components of the QGRJP. Mouse models of liver fibrosis were established by injecting olive oil containing 25% carbon tetrachloride (CCl4), which was administered at different doses of QGRJP by gavage. Liver damage and function were assessed using serum biochemical detection, ultrasound imaging, and histopathological examination. The anti-fibrosis effect was assessed using immunohistochemistry, western blotting, and quantitative real-time PCR (qRT-PCR). The in vivo safety of the QGRJP was evaluated using weight monitoring and biopsy. Potential anti-liver fibrosis signalling pathways and key targets of QGRJP were identified using RNA-seq analysis and network pharmacology. The predicted targets and pathways were validated using in vitro and in vivo experiments. ResultsQGRJP significantly ameliorated CCl4-induced liver fibrosis, and its mechanism was correlated with the inhibition of hepatic stellate cell (HSC) activation and the inflammatory response via inhibition of the TGF-β1/Smad and PI3K/AKT pathways, leading to a significant reduction in the expression of collagen and other fibrosis-related proteins. Additionally, no obvious toxic side effects were observed in the major organs of the mice or in activated HSCs (aHSCs). ConclusionThis study demonstrated that QGRJP mitigated liver injury, inflammation, and fibrosis by inhibiting the TGF-β1/Smad and PI3K/AKT signalling pathways.