绿原酸对心肌细胞炎症反应和MEK/ERK信号通路的影响

Effects of Chlorogenic Acid on MEK/ERK Signaling Pathway and Inflammatory Reaction in Cardiomyocyte

目的:探讨绿原酸对心肌细胞炎症反应和MEK/ERK信号通路的影响。方法:常规培养心肌细胞,利用脂多糖(lipopolysaccharide,LPS)诱导细胞炎症模型,将细胞分为正常对照组、LPS诱导组、LPS+50μg/mL绿原酸组、LPS+100μg/mL绿原酸组、LPS+200μg/mL绿原酸组;采用CCK8试验分析绿原酸对心肌细胞相对存活率的影响,流式细胞术检测绿原酸对心肌细胞凋亡的影响,ELISA法分析绿原酸对心肌细胞炎症反应的影响,免疫印迹法探讨绿原酸对心肌细胞MEK/ERK信号通路活化的影响。结果:LPS诱导组心肌细胞凋亡率为(52.3±4.2)%,高于正常对照组的(4.6±0.7)%;LPS诱导组炎症因子白细胞介素6(IL-6)为(1702±287)pg/mL,高于正常对照组的(529±102)pg/mL;LPS诱导组肿瘤坏死因子α(TNF-α)为(198±24)pg/mL,高于正常对照组的(81±13)pg/mL;LPS诱导组较正常对照组MEK及p-ERK蛋白表达水平显著上调(P<0.05)。与LPS诱导组相比,绿原酸可以提高细胞的存活率、抑制其凋亡、降低炎症因子IL-6和TNF-α的释放、抑制MEK/p-ERK蛋白的表达,相对于LPS诱导组差异均有统计学意义(P<0.05)。结论:绿原酸可降低LPS诱导的心肌细胞炎症反应,其机制可能与抑制MEK/ERK信号通路活化有关。

Objective:To study the effects of chlorogenic acid on MEK/ERK signaling pathway and inflammatory reaction in cardiomyocyte.Methods:After conventional culture of cardiomyocyte,LPS was used to induce cellular inflammation model,the cells were allocated to normal control group,LPS-induced group,LPS+50μg/mL chlorogenic acid group,LPS+100μg/mL chlorogenic acid group and LPS+200μg/mL chlorogenic acid group,CCK8 test was used to analyze the effects of chlorogenic acid on cardiomyocyte relative survival rate,flow cytometry was adopted to detect the effects of chlorogenic acid on the apoptosis of cardiomyocyte,ELISA method to analyze the influence of chlorogenic acid on inflammatory reaction of cardiomyocyte,Western blotting to explore the effects of chlorogenic acid on MEK/ERK signaling pathway activation.Results:Apoptosis rate of LPS-induced group was(52.3±4.2)%,higher than(4.6±0.7)%of normal control group;the concentrations of inflammatory factor IL-6 were(1702±287)pg/mL,higher than(529±102)pg/mL of normal control group;the concentrations of TNF-αin LPS-induced group were(198±24)pg/mL,higher than(81±13)pg/mL of normal control group;the expressions of MEK and p-ERK protein rose significantly in LPS-induced group compared with normal control group(P<0.05).Compared with LPS-induced group,chlorogenic acid could lift cellular survival rate,inhibit its apoptosis,reduce the release of inflammatory factor IL-6 and TNF-α,inhibit the expressions of MEK/p-ERK protein,when it was compared with LPS-induced group,the difference had statistical meaning(P<0.05).Conclusion:Chlorogenic acid could relieve LPS-induced inflammatory reaction in cardiomyocyte,and its mechanism might be related to inhibiting MEK/ERK signaling pathway activation.