Route-Dependent Proteomic Landscape in Mouse Models of Carbon Tetrachloride-Induced Hepatic Fibrosis

Hepatic fibrosis, a pathological consequence of chronic liver injury, is characterized by excessive extracellular matrix (ECM) deposition, increasing the risk of hepatocellular carcinoma. The carbon tetrachloride (CCl4)-induced mouse model is well-established for studying the pathogenesis and treatment of hepatic fibrosis, yet the effect of administration routes on fibrotic development and characteristics remains unclear. This study employed comparative proteomics to evaluate fibrosis induced via three CCl4 delivery methods: intraperitoneal (IP), subcutaneous (SC), and intragastric (IG) administration. The results demonstrated that the administration route critically determined fibrotic severity, with IG causing the most severe fibro-inflammatory injury, followed by SC and IP. Proteomic profiling identified distinct molecular pathways: IG and SC were closely associated with tissue remodeling, while IP was correlated with immune activation. Cross-species analysis further highlighted conserved profibrotic mechanisms and the hub gene patterns of hepatic ECM remodeling and metabolism differing from mice and humans. These findings underscore the importance of the CCl4 administration method as a crucial variable influencing the extent and nature of fibrosis in mouse models, and identify IG as the most effective modeling for advanced fibrosis. These insights offer a valuable foundation for refining experimental approaches to better mimic human hepatic fibrosis and enhance the translational relevance of research outcomes.