Extracellular glucose is crucially involved in the fate decision of LPS-stimulated RAW264.7 murine macrophage cells
Extracellular glucose is crucially involved in the fate decision of LPS-stimulated RAW264.7 murine macrophage cells
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DOI：
10.1038/s41598-020-67396-6
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发表时间：
2020-06-29
影响因子：
4.6
通讯作者：
Uemura, Koichi
中科院分区：
文献类型：
--
作者：
Aki, Toshihiko;Funakoshi, Takeshi;Uemura, Koichi
焦亡是一种炎性细胞死亡，依赖于炎症性半胱天冬酶介导的Gasdermin D（GSDMD）裂解，以及随后在质膜上形成孔，通过该孔，白细胞介素（IL）-1 β和IL-18从细胞中释放。在促炎激活过程中，巨噬细胞将其代谢从有氧氧化磷酸化转变为无氧糖酵解。缺氧诱导因子（HIF）1 α参与诱导IL-1 β基因表达以及代谢向糖酵解的转变。然而，焦亡和糖酵解之间的关系，以及焦亡和HIF 1 α之间的研究很少。在这里，我们表明，脂多糖（LPS）刺激的RAW 264.7小鼠巨噬细胞的细胞在细胞培养在高葡萄糖培养基中时，导致焦亡。在细胞凋亡过程中，HIF 1 α激活短暂发生，随后下调至亚基础水平。当在高糖条件下用LPS刺激细胞时，观察到HIF 1 α下调和焦亡。我们还发现，细胞内的甲基乙二醛（MGO），糖酵解的副产品，增加细胞内水平时，在高糖条件下用LPS刺激。添加糖酵解抑制剂和雷帕霉素抑制HIF 1 α下调和细胞凋亡。这些结果表明，糖酵解不仅在LPS刺激的RAW 264.7巨噬细胞的促炎活化中起关键作用，而且在细胞凋亡中也起关键作用。
Pyroptosis, a type of inflammatory cell death, is dependent on the inflammatory caspase-mediated cleavage of gasdermin D (GSDMD), and the subsequent pore formation on plasma membranes through which interleukin (IL)-1 beta and IL-18 are released from cells. During proinflammatory activation, macrophages shift their metabolism from aerobic oxidative phosphorylation to anaerobic glycolysis. Hypoxia-inducible factor (HIF)1 alpha is involved in the induction of IL-1 beta gene expression as well as the metabolic shift towards glycolysis. However, the relationships between pyroptosis and glycolysis, as well as between pyroptosis and HIF1 alpha are poorly investigated. Here we show that lipopolysaccharide (LPS) stimulation of RAW264.7 murine macrophage cells results in pyroptosis when cells are cultured in high glucose medium. During pyroptosis, HIF1 alpha activation occurs transiently followed by downregulation to sub-basal levels. HIF1 alpha downregulation and pyroptosis are observed when cells are stimulated with LPS under high glucose conditions. We also found that intracellular levels of methylglyoxal (MGO), a side product of glycolysis, increase when cells are stimulated with LPS under high glucose conditions. The addition of glycolysis inhibitor and rapamycin suppresses HIF1 alpha downregulation and pyroptosis. These results show that glycolysis plays a crucial role not only in pro-inflammatory activation, but also in pyroptosis in LPS-stimulated RAW264.7 macrophages.