基于Nrf2/HO-1通路探讨反-查耳酮对肠上皮细胞铜死亡的影响OA

Objective To investigate the effects of trans-chalcone on cuproptosis in intestinal epithelial cells,focusing on the Nrf2/HO-1 signaling pathway.Methods An in vitro ulcerative colitis(UC)-associated cuproptosis model was established by treating NCM460 cells with 30 mg/mL dextran sulfate sodium(DSS)plus copper chloride(CuCl2).After 24 h of trans-chalcone treatment,cell viability was assessed via Cell Counting Kit-8(CCK-8);intracellular copper concentration,glutathione(GSH)levels,and reactive oxygen species(ROS)production were measured using respective assay kits.Inflammatory cytokines(TNF-α,IL-1 β,IL-6)were quantified by enzyme-linked immunosorbent assay(ELISA).Western blot analysis was used to detect protein expression of cuproptosis markers(FDX1,LIAS)and Nrf2/HO-1 signaling pathway-related proteins.Results Treatment of NCM460 cells with 30 mg/mL DSS plus 30 μmol/L CuCl2 signifi-cantly reduced cell viability,accompanied by marked alterations in cuproptosis-related indicators(intracellular copper concentration,GSH levels,and ROS production),confirming successfully establishing an inflammatory cell cuproptosis model.Notably,30 μmol/L trans-chal-cone effectively restored DSS+CuCl2-induced reduction in cell viability,effectively inhibited the secretion of inflammatory cytokines,de-creased intracellular copper ions and ROS levels,and enhanced GSH levels.Furthermore,trans-chalcone modulated the protein expression of cuproptosis markers(FDX1,LIAS)and key proteins in the Nrf2/HO-1 signaling pathway induced by DSS+CuCl2(all P＜0.05).Conclu-sion Trans-chalcone may alleviate DSS and CuCl2-induced cuproptosis in UC-associated intestinal epithelial cells by regulating the Nrf2/HO-1 signaling pathway.