Wu Yong-Hong, Li Quan, Li Ping, Liu Bei
Department of Medical Technology, Xi'an Medical University, China.
Center of Stomatology, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Biochem Biophys Res Commun. 2016 Nov 18;480(3):289-295. doi: 10.1016/j.bbrc.2016.10.001. Epub 2016 Oct 3.
LPS stimulation in macrophages/monocytes induces TNFα production. We here tested the potential effect of GSK621, a novel AMP-activated protein kinase (AMPK) activator, against the process. In RAW264.7 macrophages, murine bone marrow-derived macrophages (BMDMs), and chronic obstructive pulmonary disease (COPD) patients' monocytes, GSK621 significantly inhibited LPS-induced TNFα protein secretion and mRNA synthesis. Inhibition of AMPK, through AMPKα shRNA knockdown or dominant negative mutation (T172A), almost abolished GSK621's suppression on TNFα in RAW264.7 cells. Reversely, forced-expression of a constitutively-active AMPKα (T172D) mimicked GSK621 actions and reduced LPS-induced TNFα production. Molecularly, GSK621 suppressed LPS-induced reactive oxygen species (ROS) production and nuclear factor kappa B (NFκB) activation. In vivo, GSK621 oral administration inhibited LPS-induced TNFα production and endotoxin shock in mice. In summary, GSK621 activates AMPK signaling to inhibit LPS-induced TNFα production in macrophages/monocytes.
巨噬细胞/单核细胞中的脂多糖（LPS）刺激可诱导肿瘤坏死因子α（TNFα）的产生。我们在此测试了新型AMP激活蛋白激酶（AMPK）激活剂GSK621对该过程的潜在影响。在RAW264.7巨噬细胞、小鼠骨髓来源的巨噬细胞（BMDM）以及慢性阻塞性肺疾病（COPD）患者的单核细胞中，GSK621显著抑制LPS诱导的TNFα蛋白分泌和mRNA合成。通过AMPKα短发夹RNA（shRNA）敲低或显性负突变（T172A）抑制AMPK，几乎消除了GSK621对RAW264.7细胞中TNFα的抑制作用。相反，组成型活性AMPKα（T172D）的强制表达模拟了GSK621的作用并减少了LPS诱导的TNFα产生。在分子水平上，GSK621抑制LPS诱导的活性氧（ROS）产生和核因子κB（NFκB）激活。在体内，口服GSK621可抑制小鼠中LPS诱导的TNFα产生和内毒素休克。总之，GSK621激活AMPK信号传导以抑制巨噬细胞/单核细胞中LPS诱导的TNFα产生。
