mice colon tissues RNA sequencing

Sishen Wan (SSW), a classical traditional Chinese medicine decoction, is described to treat ulcerative colitis (UC) patients, but the molecular mechanisms of the main active ingredients of SSW on the interaction between mitochondria and T cells are still unclear. This study aimed to determine the main active ingredients of SSW, predict and explore the possible regulatory mechanism of main active ingredients of SSW in modulating mitochondrial function and ameliorating mitochondrial damage, followed by regulating T cell balance during UC development. Colorimetric test and qRT-PCR showed that SSW effectively ameliorated the colonic injuries and alleviated the oxidative stress in dextran sulphate sodium (DSS)-induced UC mice. Network pharmacology, molecular docking, dynamic simulation and 3D-QSAR exhibited that angelicin, corylifolinin, psoralen and rutaecarpine, derived from SSW, were identified as the main components of SSW, which might interact with CYP2C9 and CYP1A1 due to the lowest binding energy. RNA-seq analysis and western blot (WB) exhibited that SSW alleviated UC via regulating genes related to mitochondrial function and T cell responses based on RNA-seq data. Cytc-related targets and T cell-associated proinflammatory cytokines were downregulated, while mtDNA repairing-related targets were upregulated with SSW intervention. Moreover, the caspase, inflammasome and Th1 and Th17 polarizing-related genes are positively correlated with COX, caspase and inflammasome-associated genes, respectively. Taken together, this study not only identifies the main bioactive ingredients of SSW and their possible ligands, but also provides angelicin, corylifolinin, psoralen and rutaecarpine may alleviate oxidative stress and mitochondrial damage, followed by modulating Th1 and Th17-related proinflammatory cytokines.