摘要
目的:探讨葛根素(Puerarin)能否通过抑制炎症反应,调控SLC7A11/GPX4信号轴介导的铁死亡途径改善脓毒症心肌细胞损伤.方法:采用脂多糖(LPS)刺激H9c2大鼠心肌细胞构成脓毒症心肌损伤细胞模型,采用空白对照(Control)组、脓毒症(LPS)组、脓毒症+葛根素(LPS+Pue)组.通过试剂盒检测心肌细胞活力,通过ELISA检测白细胞介素(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor,TNF-α)含量,通过试剂盒测定氧化应激指标谷胱甘肽(Glutathione,GSH)、丙二醛(Malondialdehyde,MDA)变化水平,通过Western blot检测调控铁死亡相关蛋白,如溶质载体家族成员(Solute Carrier Family 7,SLC7A11)、谷胱甘肽过氧化物酶4(Glutathione peroxidase 4,GPX4)的表达变化.结果:相较于Control组,在LPS刺激后,H9c2细胞活性降低,MDA水平上升,炎症因子增多,铁死亡调控蛋白SLC7A11、GPX4的表达水平下降(P＜0.05).相较于LPS组,在Pue给药处理后,随Pue浓度的增加细胞活性随之升高,呈剂量依赖性,同时,细胞炎症水平随之降低,铁死亡蛋白SLC7A11、GPX4的表达水平也随之上升(P＜0.05).结论:葛根素可通过上调SLC7A11/GPX4信号通路,减轻细胞内脂质过氧化,抑制铁死亡进程,进而改善LPS诱导的心肌细胞损伤.
Abstract
Objective:To investigate whether Puerarin can regulate SLC7A11/GPX4 signaling axis-mediated ferroptosis pathway to improve myocardial cell damage in sepsis by inhibiting inflammatory response.Methods:H9c2 rat cardiomyocytes were induced by LPS to form a model of septic myocardial damage cells,which were divided into Control group,sepsis(LPS)group and sepsis+puerarin(LPS+Pue)group.Cardiomyocyte viability by kit,interleukin(IL-6)and tumour necrosis factor-α(TNF-α)levels by ELISA,oxidative stress indicators glutathione(GSH)and malondialdehyde(MDA)levels by kit,and expression of regulatory ferroptosis-related proteins SLC7A11 and GPX4 by Western blot.Results:Compared to the Control group,after LPS stimulation,H9c2 cell activity decreased,MDA levels increased,inflammatory factors increased and the expression levels of ferroptosis regulatory proteins SLC7A11 and GPX4 decreased(P＜0.05).Compared to the LPS group,after Pue administration treatment,cellular activity decreased with increasing Pue concentration in a dose-dependent manner,while cellular inflammation levels decreased and the expression levels of ferroptosis proteins SLC7A11 and GPX4 increased(P＜0.05).Conclusion:Puerarin reduces LPS-induced inflammatory factors and oxidative stress levels in H9c2 cells by upregulating the SLC7A11/GPX4 signaling pathway,attenuating intracellular lipid peroxidation,and inhibiting ferroptosis.
基金项目
省部共建中亚高发病成因与防治国家重点实验室青年基金(SKL-HIDCA-2023-YY12)
维普
万方数据