人参皂苷Rb1通过抑制PI3K/AKT信号通路保护脂多糖诱导的心肌细胞炎症反应

Protective effect of ginsenoside Rb1 on LPS-induced inflammatory response of cardiomyocytes through inhibiting PI3K/AKT signal pathway

目的探究人参皂苷Rb1在脂多糖(LPS)诱导的H9C2心肌细胞炎症反应中的作用及分子机制。方法建立LPS诱导的H9C2心肌细胞炎症反应模型。在用1μg/ml的LPS诱导H9C2心肌细胞炎症反应前,使用100μM人参皂苷Rb1或1μM PI3K抑制剂Wortmannin预处理1 h,2 h后收集细胞样本。利用q PCR检测白介素-6(IL-6),白介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)的m RNA表达水平;用ELISA试剂盒检测细胞上清IL-6,IL-1β,TNF-α的蛋白表达;用WB实验检测p-AKT和AKT的表达水平。结果人参皂苷Rb1显著抑制LPS诱导的IL-6,IL-1β,TNF-α的m RNA(P<0.001)和蛋白(P<0.001)表达水平;人参皂苷Rb1显著抑制LPS诱导的PI3K/AKT信号通路的激活(P<0.001);PI3K抑制剂Wortmannin也能显著抑制LPS诱导的p-AKT表达水平以及IL-6,IL-1β,TNF-α的m RNA(P<0.001)和蛋白(P<0.001)表达水平。结论人参皂苷Rb1通过抑制PI3K/AKT信号通路阻止LPS诱导的心肌细胞炎症反应。

Objective To study the effect and molecular mechanism of ginsenoside Rb1 in inflammatory response of H9C2 cardiomyocytes induced by lipopolysaccharide(LPS).Methods The model of inflammatory response of H9C2 cardiomyocytes induced by LPS was established.Before reducing inflammatory response with LPS(1μg/mL),H9C2 cardiomyocytes were pretreated with ginsenoside Rb1(100μM)or Wortmannin(PI3K inhibitor,1μM)for 1 h,and cell samples were collected after 2 h.The mRNA expressions of interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The protein expressions of IL-6,IL-1βand TNF-αwere detected by using ELISA,and expressions of p-AKT and AKT were detected by using Western blotting assay.Results Ginsenoside Rb1 inhibited significantly mRNA expressions(P<0.001)and protein expressions(P<0.001)IL-6,IL-1βand TNF-αinduced by LPS.Ginsenoside Rb1 inhibited significantly the activation of PI3K/AKT signal pathway induced by LPS(P<0.001).Wortmannin also inhibited significantly p-AKT expression,and mRNA expressions(P<0.001)and protein expressions(P<0.001)IL-6,IL-1βand TNF-αinduced by LPS.Conclusion Ginsenoside Rb1 can stop the inflammatory response of cardiomyocytes induced by LPS through inhibiting PI3K/AKT signal pathway.