基础医学与临床 ›› 2015, Vol. 35 ›› Issue (7): 879-883.

• 研究论文 • 上一篇    下一篇

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

杨蕾,李蓉,李岱,敬媛媛,杨利,陶安阳,周庆菊,赵红燕   

  1. 重庆医科大学附属第一医院
  • 收稿日期:2015-01-04 修回日期:2015-04-23 出版日期:2015-07-05 发布日期:2015-06-23
  • 通讯作者: 李蓉 E-mail:rongli232006@163.com
  • 基金资助:
    国家自然科学基金项目;国家临床重点专科建设项目

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

  • Received:2015-01-04 Revised:2015-04-23 Online:2015-07-05 Published:2015-06-23

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<.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。

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, 炎症因子

Abstract: 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 + SFA-treated group and AMPK inhibitor Compound C + metformin + SFA-treated group. Real-time quantitative 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 blotting was used to analyze phosphorylation level of AMP-activated protein kinase. Results Compared to normal control group, both mRNA expression levels and protein secretion levels of TNF-α and IL-6 were elevated in SFA-treated group (P<0.05). Compared to SFA-treated group, both mRNA expression levels and protein secretion levels of TNF-α and IL-6 were 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 levels of TNF-α and IL-6 were elevated, and AMPK phosphorylation level was deceased in Compound C + metformin + SFA-treated group (P<0.05). Conclusion Metformin suppresses secretion of inflammatory cytokines TNF-α and IL-6 in saturated fatty acid-induced RAW264.7 macrophage via activation AMPK.

Key words: RAW264.7 macrophage, metformin, saturated fatty acid, AMP-activated protein kinase, inflammatory cytokine

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