二甲双胍对饱和脂肪酸诱导的大鼠H9C2型心肌细胞损伤的保护作用研究

目的 探讨二甲双胍(Met)对饱和脂肪酸所诱导的大鼠H9C2型心肌细胞损伤作用的保护机制.方法 在对照、棕榈酸(PA)及3种不同浓度梯度Met与PA联合组培养液中培养大鼠H9C2型心肌细胞株24 h.采用蛋白质印迹法(Western blot)测定各组细胞核因子κB(NF-κB)p65、细胞间黏附因子(ICAM1)、磷酸化核因子κB抑制蛋白(p-IκBα)及磷酸化腺苷酸活化蛋白激酶(p-AMPK)的蛋白表达,实时荧光定量PCR测定各组细胞NF-κB、单核细胞趋化因子(CCL2)和ICAM1的mRNA表达.结果 与对照组相比,PA组的细胞中NF-κB p65、ICAM1、p-IκBα蛋白表达量增加(P<0.05);与PA组相比,Met+PA联合组细胞NF-κB p65、ICAM1、p-IκBα的蛋白表达量随Met浓度梯度增加表现为不同程度递减(P<0.05),p-AMPK蛋白表达量随Met浓度增加而显著增加(P<0.05).与对照组相比,PA组CCL2、ICAM1的mRNA表达量增加(P<0.05);与PA组相比,Met+PA联合组细胞CCL2和ICAM1的mRNA表达量下降(P<0.05).结论 Met可以减轻由饱和脂肪酸诱导细胞黏附因子和趋化因子表达增加而引起的大鼠H9C2型心肌细胞损伤.

Protective effect of metformin on H9C2 rat myocardial cell damage induced by saturated fatty acid

Objective To investigate the mechanisms of metformin (Met) for protecting H9C2 myocardial cells damage induced by saturated fatty acids (palmitic acid,PA) in rats.Methods Rat H9C2 myocardial cell lines in blank control group,PA group and three different concentrations of metformin and PA combination groups were cultured for 24 h.Then Western blot was adopted to detect the protein expression of nuclear factor κB p65 (NFκB p65),intercellular cell adhesion molecule-1 (ICAM1),phosphorylated inhibitor of nuclear factor κB (p-IκBα) and phosphorylated adenosine monophosphate activated protein kinase(p-AMPK);the quantitative real-time PCR(qRT-PCR) was used to detect the mRNA expression of nuclear factor of nuclear factor κB(NFκB),chemokine (C-C motif) ligand 2 (CCL2) andintercellular cell adhesion molecule-1(ICAM1).Results Compared with the control groups,the protein expression of NFκB p65,p-IκBα and ICAM1 in the PA group was increased (P<0.05);compared with the PA group,the protein expression of NFκB p65,IκBα and ICAM1 in the Met+PA combination groups was decreased in various degrees with the Met concentration increase (all P<0.05),while the protein expression of p-AMPK was significantly risen with the Met concentration increase (all P<0.05).Compared with the control group,the mRNA expression of CCL2 and ICAM1 in the PA group was increased (P<0.05);compared with the PA group,the mRNA expression of CCL2 and ICAM1 in the Met+PA combination groups was decreased (P<0.05).Conclusion Met can alleviate H9C2 myocardial cellular damage caused by saturated fatty acid inducing increase of CCL2 and ICAM1 expression.