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.肝纤维化（hepatic fibrosis）是慢性肝损伤的病理性结局，以细胞外基质（extracellular matrix，ECM）过度沉积为特征，会增加肝细胞癌的发病风险。四氯化碳（carbon tetrachloride，CCl4）诱导的小鼠模型是研究肝纤维化发病机制与治疗策略的经典模型，但给药途径对纤维化发生发展及病理特征的影响仍未明确。本研究采用比较蛋白质组学方法，评估了三种CCl4给药方式：腹腔注射（intraperitoneal，IP）、皮下注射（subcutaneous，SC）与灌胃（intragastric，IG）诱导的肝纤维化情况。研究结果表明，给药途径是决定纤维化严重程度的核心变量：灌胃给药引发的纤维化炎症损伤最为严重，其次为皮下注射与腹腔注射。蛋白质组谱分析鉴定出了差异化的分子通路：灌胃与皮下注射给药模型与组织重构紧密相关，而腹腔注射给药模型则与免疫激活存在关联。跨物种分析进一步揭示了保守的促纤维化机制，且小鼠与人类的肝ECM重构及代谢核心基因模式存在显著差异。本研究结果强调，CCl4给药途径是影响小鼠模型纤维化程度与病理特征的关键变量，并证实灌胃给药是构建晚期纤维化模型的最优方式。这些研究发现为优化实验方案以更好地模拟人类肝纤维化、提升研究成果的转化相关性提供了极具价值的理论基础。