Integration of network pharmacology, lipidomics, and transcriptomics analysis to reveal the mechanisms underlying the synergy between susceptibility factor TNF-α and Polygonum multiflorum-induced idiosyncratic liver injury in mice.

Idiosyncratic Drug-Induced Liver Injury (IDILI) challenges drug development and clinical use. Its unpredictable nature complicates mechanistic research. Polygonum multiflorum Thunb (PM), a traditional tonic herb, has been used for centuries in China and East Asia. Now, PM-induced IDILI is a focus of traditional Chinese medicine safety research. Nevertheless, the underlying mechanisms of PM-triggered IDILI require further investigation. Previous clinical studies identified elevated Tumor Necrosis Factor-alpha(TNF-α) levels in patients susceptible to PM-induced liver injury. This finding prompted us to employ integrated network pharmacology, targeted lipidomics, and transcriptomics to investigate the combined effect and mechanism of PM and the susceptibility factor TNF-α in C57 mice. Histochemical staining, biochemical assays, and inflammatory cytokine analysis revealed significant hepatic inflammatory infiltration in the TNF-α+PM co-exposure group versus controls. This group exhibited markedly elevated levels of liver injury markers (ALT, AST, DBIL) and inflammatory mediators (IL-6, TNF-α) (P<0.05, P<0.01). Subsequent administration of the TNF-α antagonist etanercept restored hepatic architecture and significantly reduced cytokine levels (P<0.01), approaching baseline values. Notably, neither TNF-α nor PM monotherapy induced liver injury in mice. Network pharmacology analysis identified 33 bioactive PM components and 11 core targets. Targeted lipidomics revealed 32 differential lipid metabolites (DLMs), primarily glycerolipids (GL) and glycerophospholipids (GP). Transcriptomics demonstrated cooperative regulation of 199 differentially expressed genes(DEGs) by TNF-α and PM. Network enrichment analysis further uncovered lipid dysregulation mediated by Pparg, Cyp4a12a, Cyp4a12b, and Cyp4a31 genes, with RT-qPCR validation confirming these findings. In summary, TNF-α plays a central driving role in PM-induced liver injury. When co-administered with PM, it activates the PPAR signal ing pathway via downregulation of Cyp4a12a, Cyp4a12b, and Cyp4a31, thereby dysregulating lipid metabolism, promoting inflammatory mediator production, and ultimately causing liver injury.