Main active components of Sishen Wan may modulate T cells-related proinflammatory cytokines via alleviating mitochondrial damage caused by oxidative stress in dextran sulphate sodium-induced ulcerative colitis

Introduction

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.

Methods

Colorimetric test and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the efficacy of SSW on inflammatory injuries of UC and preliminarily explore the mechanisms of SSW against oxidative stress. The main active components and their possible ligands were predicted by network pharmacology, molecular docking, dynamic simulation and three-dimensional-quantitative structure activity relationship (3D-QSAR). RNA-seq analysis and western blot (WB) was conduct to discover the impact of SSW on genetic profile changes, and discover and predict the potentials of anti-mitochondrial damage and proinflammatory T-cells of the selected bioactive compounds.

Results

SSW effectively ameliorated the colonic injuries and alleviated the oxidative stress in the dextran sulphate sodium (DSS)-induced UC. Angelicin, corylifolinin, psoralen and rutaecarpine, derived from SSW, were identified as the main components of SSW, and might interact with CYP2C9 and CYP1A1 due to the lowest binding energy. 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 cytochrome C oxidase (COX), caspase and inflammasome-associated genes, respectively.

Conclusion

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.