Background: Traditional Chinese medicine bear bile has shown significant efficacy in improving nonalcoholic steatohepatitis (NASH) ; however, its use raises ethical concerns regarding animal welfare, creating an urgent need for alternatives. Cultured bear bile powder (CBBP) has emerged as a promising natural substitute because of its core compositional similarities, comparable therapeutic effects, and a more sustainable sourcing process that better aligns with animal protection principles. Systematic investigation of the efficacy and underlying mechanisms of CBBP against NASH is therefore of considerable significance. Objective: This study aimed to investigate the therapeutic effects and underlying mechanisms of CBBP in NASH. Methods: In this study, a NASH model was established in male C57BL/6 mice using a Western diet combined with carbon tetrachloride to explore the protective effects and underlying mechanisms of CBBP. The therapeutic effects of CBBP were systematically evaluated through liver-to-body weight ratio, the nonalcoholic fatty liver disease activity score, histopathological observations, and the levels of the liver injury markers and inflammatory factors. A comprehensive approach was employed to elucidate the underlying mechanisms, combining untargeted lipidomic analysis, network pharmacology, enzyme-linked immunosorbent assay, and immunohistochemistry staining to identify the key genes and pathways involved. Results: Treatment with CBBP effectively ameliorated liver pathological injury, suppressed liver fibrosis, reduced hepatic lipid accumulation, and decreased liver injury markers and inflammatory factors levels in both serum and liver of NASH mice. Untargeted lipidomic analysis revealed that the effects of CBBP were primarily associated with triglycerides (TGs) and phosphatidylcholines, significantly decreasing TG (74: 3) and TGs (18: 1₁8: 1₂3: 1) while increasing phosphatidylcholine (18: 0₂2: 6). Pathway enrichment analysis showed that CBBP modulated choline metabolism, glycerophospholipid metabolism, and cholesterol metabolism to improve lipid metabolic abnormalities in NASH. The underlying mechanisms and potential molecular targets of CBBP in regulating lipid metabolism were further investigated based on network pharmacology. The Kyoto Encyclopedia of Genes and Genomes results and subsequent validation experiments demonstrated that CBBP inhibited activation of the phosphatidylinositol 3-kinase-protein kinase B signaling pathway, which may contribute to its therapeutic effects in NASH. Conclusion: In summary, our study confirms the therapeutic potential of CBBP in NASH and provides valuable insights into its underlying mechanisms through combined lipidomic and network pharmacology analysis. The phosphatidylinositol 3-kinase-protein kinase B signaling pathway may play a key role in mediating the beneficial effects of CBBP in NASH.
Li et al. (Wed,) studied this question.