二甲双胍激活AMPK通路减轻饱和脂肪酸诱导的RAW264.7巨噬细胞炎性反应  

Metformin alleviates saturated fatty acid-induced inflammation in RAW264.7 macrophage via activating AMPK pathway

作　　者：杨蕾[1] 李蓉[1] 李岱[1] 敬媛媛[1] 杨利[1] 陶安阳 周庆菊[1] 赵红燕[1] 

机构地区：[1]重庆医科大学附属第一医院内分泌科,重庆400016

出　　处：《基础医学与临床》2015年第7期879-883,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81200588);国家临床重点专科建设项目(2011)

摘　　要：目的观察二甲双胍对饱和脂肪酸(SFA)诱导的RAW264.7巨噬细胞炎性反应中炎性因子的影响及探讨其可能机制。方法 SFA干预RAW264.7巨噬细胞建立体外炎性反应模型;实验分为对照组、SFA干预组、二甲双胍+SFA干预组、AMPK抑制剂Compound C+二甲双胍+SFA干预组;实时定量PCR检测TNF-α和IL-6 mRNA的表达,ELISA法检测TNF-α和IL-6蛋白的分泌,Western blot分析腺苷酸活化蛋白激酶(AMPK)的磷酸化水平。结果与对照组比较,SFA干预组RAW264.7巨噬细胞TNF-α、IL-6 mRNA表达及蛋白分泌水平均显著升高(P<0.05);与SFA组比较,二甲双胍+SFA干预组TNF-α、IL-6 mRNA表达水平及蛋白分泌水平均降低(P<0.05),而细胞AMPK的磷酸化水平增强(P<0.05);与二甲双胍+SFA干预组比,Compound C+二甲双胍+SFA干预组TNF-α、IL-6 mRNA表达及蛋白分泌水平均升高,AMPK磷酸化水平降低(P<0.05)。结论二甲双胍激活AMPK降低饱和脂肪酸诱导的RAW264.7巨噬细胞炎性因子TNF-α、IL-6的分泌。Objective To investigate the effect of metformin on inflammatory cytokines of saturated fatty acid （SFA）-induced RAW264.7 macrophage.Methods RAW264.7 cells were treated with SFA to establish inflammatory model in vitro.Macrophages were divided into normal control group,SFA-treated group,metformin ＋ SFAtreated group and AMPK inhibitor Compound C ＋ metformin ＋ SFA-treated group.RT-PCR was used to detect mRNA expression levels of TNF-α and IL-6.ELISA was used to detect secretion levels of TNF-α and IL-6 in cell supernatants.Western blot was used to analyze phosphorylation level of AMP-activated protein kinase.Results Compared to normal control group,both mRNA expression and protein secretion of TNF-α and IL-6 increased in SFA-treated group （P ＜ 0.05）.Compared to SFA-treated group,both mRNA expression and protein secretion of TNF-α and IL-6 decreased in metformin ＋ SFA-treated group （P ＜0.05）,and AMPK phosphorylation level was significantly enhanced （P ＜ 0.05）.Compared to metformin ＋ SFA treated-group,mRNA expression and protein secretion of TNF-α and IL-6 increased,and AMPK phosphorylation level deceased in Compound C ＋ metformin ＋SFA-treated group （P ＜ 0.05）.Conclusions Metformin suppresses secretion of inflammatory cytokines TNF-αand IL-6 in saturated fatty acid-induced RAW264.7 macrophage via activation of AMPK.

关 键 词：RAW264.7巨噬细胞 二甲双胍 饱和脂肪酸 腺苷酸活化蛋白激酶 炎性因子 

分 类 号：R589.2[医药卫生—内分泌]