Multi-omics reveals rutin directly targets RUNX1 to disrupt the RUNX1/TET2 complex and alleviate NAFLD via TLR4/NF-κB inhibition.

Non-alcoholic fatty liver disease (NAFLD) is a prevalent liver disorder driven by metabolic dysregulation and chronic inflammation, for which targeted pharmacotherapies remain limited. Rutin, a bioactive flavonoid from Sophora japonica and Fagopyrum esculentum, possesses notable anti-inflammatory and antioxidant properties. This study explored its pharmacological effects and underlying mechanism in NAFLD using a combination of in vivo and in vitro approaches. We found that rutin administration markedly attenuated hepatic steatosis, reduced oxidative stress, restored mitochondrial function, and improved liver injury markers, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), in both high-fat diet (HFD)-fed ApoE-/- mice and free fatty acid (FFA)-exposed HepG2 cells. Furthermore, rutin significantly suppressed the production of pro-inflammatory cytokines, including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Mechanistic studies integrating multi-omics and molecular biology approaches demonstrated that rutin directly binds to Runt-related transcription factor 1 (RUNX1), disrupts its interaction with ten-eleven translocation 2 (TET2), and thereby inhibits the downstream Toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway. Our results illuminate a novel pharmacological axis for rutin, positioning it as a promising multi-target candidate for NAFLD treatment by synchronously ameliorating lipid metabolism, oxidative injury, and inflammatory response.