从miR-143角度探讨黄芩汤抑制溃疡性结肠炎细胞铁死亡的作用机制研究OA

Objective To investigate whether Huangqin Decoction(黄芩汤,HQD)inhibits ferroptosis in ulcerative colitis(UC)by regulating miR-143 expression.Methods The study comprised two parts.In part one,Caco-2 cells were transfected with miR-143 mimic or miR-143 inhibitor.Both parts utilized Erastin to induce a ferroptosis model.In part two,cells were additionally treated with varying concentrations of HQD-containing serum,using Fer-1 as a control.Cell death was assessed by flow cytometry.LDH release,GSH and iron levels were measured using biochemical assays.Cell viability was determined by the CCK-8 assay.Intracellular reactive oxygen species(ROS)were measured using the DCFH-DA fluorescent probe.Expression levels of miR-143,GPX4,FTH1,and PTGS2 mRNA were quantified by qRT-PCR.Protein levels of GPX4,FTH1,and PTGS2 were analyzed by Western blot(WB).Results In part one,compared to the model group,the miR-143 mimic group showed significantly decreased cell death rate and iron levels(P<0.01),significantly increased GSH levels(P<0.01),significantly elevated GPX4 mRNA expression(P<0.01)and increased GPX4 protein levels(P<0.05),as well as significantly reduced PTGS2 mRNA and protein expression(P<0.01).Conversely,the miR-143 inhibitor group exhibited trends opposite to those observed in the mimic group.In part two,compared to the model group,both the medium-dose HQD group and the Fer-1 group demonstrated significantly increased cell viability(P<0.01),significantly decreased LDH release and ROS levels(P<0.01),significantly elevated mRNA and protein expression of GPX4 and FTH1(P<0.01),and significantly reduced mRNA and protein expression of PTGS2(P<0.01).Additionally,in part two,miR-143 expression was significantly increased in the medium/high-dose HQD groups compared to the model group(P<0.05).Conclusion HQD may inhibit ferroptosis in UC by regulating miR-143 expression.