苦杏仁苷调节HMGA1/NF-κB信号通路对LPS诱导的心肌细胞炎症反应的影响

Effect of Amygdalin on LPS-induced Myocardial Inflammation by Regulating HMGA1/NF-κB Signaling Pathway

目的:探讨苦杏仁苷(AMG)对脂多糖(LPS)诱导的心肌细胞炎症反应的影响及对高迁移率族ATHook蛋白1(HMGA1)/核因子-κB(NF-κB)信号通路的调节机制。方法:将H9c2心肌细胞分为对照组、LPS组、AMG低剂量组、AMG高剂量组、AMG高剂量+重组HMGA1组(AMG高+HMGA1组)。采用四甲基偶氮唑蓝(MTT)检测心肌细胞的增殖活性;流式细胞术检测心肌细胞凋亡率;酶联免疫吸附法(ELISA)检测细胞培养液炎性因子白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α;蛋白免疫印迹法(Western Blot)检测心肌细胞HMGA1/NF-κB通路相关蛋白的表达。结果:与对照组比较,LPS组心肌细胞存活率、NF-κB抑制蛋白α(IκBα)蛋白表达降低,心肌细胞凋亡率、炎性因子IL-6、IL-1β、TNF-α含量、HMGA1蛋白和NF-κB p65、IκBα磷酸化水平升高(P<0.05)。与LPS组比较,AMG各剂量组心肌细胞存活率、IκBα蛋白表达升高,心肌细胞凋亡率、炎性因子IL-6、IL-1β、TNF-α含量、HMGA1蛋白和NF-κB p65、IκBα磷酸化水平降低(P<0.05),且HMGA1过表达可逆转AMG对心肌细胞的保护作用(P<0.05)。结论:AMG通过抑制HMGA1/NF-κB信号通路,减轻LPS诱导的心肌细胞炎症反应。

Objective:To investigate the effect of amygdalin(AMG)on lipopolysaccharide(LPS)-induced myocardial inflammation and its regulatory mechanism of high mobility ATHook protein 1(HMGA1)/nuclear factor-κB(NF-κB)signaling pathway.Methods:H9c2 cardiomyocytes were divided into control group,LPS group,AMG low-dose group,AMG high-dose group,AMG high-dose+recombinant HMGA1 group(AMG high+HMGA1 group).The proliferative activity of cardiomyocytes was detected by tetramethylazolium blue(MTT).Cardiomyocyte apoptosis rate was detected by flow cytometry.The inflammatory factors interleukin(IL)-6,IL-1βand tumor necrosis factor(TNF)-αwere detected by enzyme-linked immunosorbent assay(ELISA).The expressions of HMGA1/NF-κB pathway related proteins in cardiomyocytes were detected by Western Blot.Results:Compared with the control group,the survival rate of cardiomyocytes and the expression of NF-κB inhibitory proteinα(IκBα)decreased in the LPS group,while the apoptosis rate,inflammatory factor IL-6,IL-1β,TNF-αcontent,HMGA1 protein and phosphorylation levels of NF-κB p65 and IκBαincreased(P<0.05).Compared with the LPS group,the survival rate of cardiomyocytes and the expression of IκBαprotein increased in AMG dose groups,while the apoptosis rate,inflammatory factor IL-6,IL-1β,TNF-αcontent,HMGA1 protein and phosphorylation levels of NF-κB p65 and IκBαdecreased(P<0.05).Overexpression of HMGA1 could reverse the protective effect of AMG on cardiomyocytes(P<0.05).Conclusion:AMG could attenuate LPS-induced myocardial inflammation by inhibiting the HMGA1/NF-κB signalling pathway.