Abstract：
Toll-like receptors 3 (TLR3) are innate immune receptors expressed on a wide range of cell types, including glial cells. Inflammatory responses altered by hyperglycemia highlight the need to explore the molecular underpinnings of these changes in cellular models. Therefore, here we estimated TLR3-mediated response of astrocytes cultured at normal (NG, 5 mM) and high (HG, 22.5 mM) glucose concentrations for 48 h before stimulation with polyinosinic:polycytidylic acid Poly(I:C) (PIC) for 6 h. Seahorse Extracellular Flux Analyzer (Seahorse XFp) was used to estimate the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Although adaptation to HG affected ECAR and OCR, the stimulation of cells with PIC had no effect on ECAR. PIC reduced maximal OCR, but this effect disappeared upon adaptation to HG. PIC-stimulated release of cytokines IL-1β, IL-10 was reduced, and that of IL-6 and iNOS was increased in the HG model. Adaptation to HG reduced PIC-stimulated synthesis of COX-derived oxylipins measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Adaptation to HG did not alter PIC-stimulated p38 activity, ERK mitogen-activated protein kinase, STAT3 and ROS production. Metformin exhibited anti-inflammatory activity, reducing PIC-stimulated synthesis of cytokines and oxylipins. Cell adaptation to high glucose concentration altered the sensitivity of astrocytes to TLR3 receptor activation, and the hypoglycemic drug metformin may exert anti-inflammatory effects under these conditions.
摘要：
Toll样受体3（TLR3）是在多种细胞类型上表达的先天免疫受体，包括神经胶质细胞.高血糖改变的炎症反应凸显了在细胞模型中探索这些变化的分子基础的必要性。因此,在这里，我们估计了正常培养的星形胶质细胞的TLR3介导的反应（NG，5mM）和高（HG，22.5mM）葡萄糖浓度48小时，然后用聚肌苷酸：聚胞苷酸聚（I：C）（PIC）刺激6小时。海马细胞外通量分析仪（海马XFp）用于估计细胞外酸化率（ECAR）和耗氧率（OCR）。虽然适应HG会影响ECAR和OCR，PIC刺激细胞对ECAR无影响。PIC降低了最大OCR，但是这种效果在适应HG后消失了。PIC刺激细胞因子IL-1β的释放，IL-10减少，HG模型中IL-6和iNOS的含量增加。通过超高效液相色谱-串联质谱（UPLC-MS/MS）测量的对HG的减少的PIC刺激的COX衍生的氧化脂素的合成。对HG的适应不会改变PIC刺激的p38活性，ERK丝裂原活化蛋白激酶,STAT3和ROS产生。二甲双胍表现出抗炎活性，减少PIC刺激的细胞因子和氧化脂素的合成。细胞对高葡萄糖浓度的适应改变了星形胶质细胞对TLR3受体激活的敏感性,降血糖药物二甲双胍在这些条件下可能发挥抗炎作用。
