Integrated single-cell transcriptomics and proteomics elucidate the molecular mechanisms and detoxification strategy of rifampicin-induced hepatotoxicity

Rifampicin (RIF), a cornerstone drug in tuberculosis treatment, is associated with hepatotoxicity, which represents a significant adverse effect that frequently causes discontinuation of therapy. However, a comprehensive evaluation of the mechanisms underlying RIF-induced hepatotoxicity remains limited, and the identification of highly effective, low-toxicity therapeutic interventions is urgently needed. In this study, we employed a RIF-induced mouse hepatotoxicity model to systematically investigate the cellular and molecular events associated with RIF-induced liver injury. By integrating single-cell RNA sequencing, bulk RNA-seq, and mass spectrometry-based proteomics and metabolomics, we identified region-specific hepatocyte damage characterized by elevated reactive oxygen species (ROS) levels and activation of the fatty acid oxidation pathway. At the molecular level, RIF treatment resulted in the upregulation of pregnane X receptor (PXR) and <i>Cyp3a11</i>, along with the downregulation of key antioxidant genes. Moreover, decreased mTOR expression and increased expression of fatty acid oxidation-related genes including <i>Acox1</i> and <i>Acaa1b</i> suggested an enhanced oxidative metabolism. Recruitment of macrophages further exacerbated hepatocyte damage. Importantly, <i>Rosmarinic acid</i> (RA) administration was shown to attenuate RIF-induced hepatotoxicity. These findings provide a comprehensive molecular and cellular perspective on RIF-induced hepatotoxicity and suggest the potential clinical application of RA as a therapeutic agent in the management of RIF-induced liver injury.