摘要
Abstract
Objective:To explore the regulatory effect of eicosapentaenoic acid(EPA)on lipopolysaccharide(LPS)-induced HBZY-1 cell pyroptosis based on the Nrf2/NLRP3/GSDMD signaling pathway.Additionally,nuclear factor E2 related factor 2(Nrf2)agonist(4-OI)and nucleotide-bound oligomerized domain-like receptors(NLRP3)receptor inhibitor(MCC950)were chosen to explore the regulating mechanism of EPA on Nrf2.Methods:According to different drug treatments strategy,HBZY-1 cells were divided into five groups:control group,LPS group,LPS+EPA group,LPS+EPA+4-OI group,and LPS+EPA+MCC950 group,in which 10 μg/mL LPS was applied to induce the cell pyroptosis in vitro,and the concentration of EPA,4-OI,MCC950 was all 200 µmol/L,the drugs were added to the culture plate and treated for 48 h.In addition,CCK-8 assay,LDH release test,and TUNEL staining were performed to detect cell damage.The release of inflammatory factors IL-1β and IL-18 in the cell supernatant were detected by ELISA kits,protein levels of Nrf2,NLRP3,Caspase-1 and GSDMD were detected by Western Blotting.Results:Cell viability assay showed that 10 μg/mL LPS stimulation for 48 hours significantly reduced the cell viability(P＜0.05),but 20-200 μmol/L EPA had significantly protective effects(P＜0.05).Moreover,200 μmol/L EPA significantly attenuated the cell damage caused by LPS(P＜0.01),reduced the release of inflammatory factors(P＜0.01),LDH release(P＜0.01)and DNA fragmentation,inhibited the activation of the pyroptosis,and promoted the expression of Nrf2 protein(P＜0.01).In addition,co-treatment with 4-OI and MCC950 found that 4-OI can promote the inhibitory effect of EPA on pyroptosis(P＜0.05),reduce the expression of related proteins and the release of IL-1 β and IL-18(P＜0.05),and enhance the protection effect of EPA(P＜0.01),while MCC950 has no significant on the effect of EPA.Conclusion:EPA inhibits the expression of various molecules in the pyroptosis of HBZY-1 cells induced by LPS through Nrf2,and exerts a significant cytoprotective effect.关键词
二十碳五烯酸/焦亡/细胞损伤/核因子E2相关因子2(Nrf2)Key words
eicosapentaenoic acid(EPA)/pyroptosis/cell damage/nuclear factor E2 related factor 2(Nrf2)分类
医药卫生
