目的观察TRL3激动剂poly(I:C)-LMW预处理对原代皮质神经元Toll样受体3(TLR3)表达的影响,探讨TLR3在缺糖缺氧诱导的原代皮质神经元损伤中的作用。方法取体外培养7d的大鼠原代皮质神经元细胞,对细胞分别予以正常条件下培养(空白对照组);缺糖缺氧2h,复糖复氧24h(OGD组);TRL3激动剂预处理12h(激动剂组);TRL3激动剂预处理12h后缺糖缺氧2h,复糖复氧24h(激动剂+OGD组)。免疫荧光法观察各组细胞生长状态,分别以Western blot法、RT-PCR法测定TLR3蛋白及TLR3mRNA的表达情况。结果 与空白对照组相比,激动剂及缺氧复氧处理方法均诱导原代皮质神经元TLR3、TLR3mRNA表达增强(P〈0.05);激动剂预处理后,与OGD组相比,激动剂+OGD组神经元细胞状态较好,数目较多(P〈0.05)。结论 TLR3参与缺糖缺氧诱导的原代皮质神经元的损伤过程,激动剂可减轻神经元缺糖缺氧后损伤。
Objective To explore whether Toll like receptors 3(TLR3) is involved in injured process which oxygen-glucose deprivation(OGD)induced primary cortical neuron,and the affection of pretreated by poly(I:C)-LMW(the agonist of TLR3) for its expression.Methods Primary cortical neuron was pretreated with poly(I:C)-LMW,it was induced by OGD after 12 hours(oxygen and glucose recovered after they was deprivated 2 hours and this continued 24 hours,the changes in the protein expression of TLR3 were detected by Western blot,the changes in the mRNA expression of TLR3 were detected by RT-PCR.The state of cells was observed by immunofluorescence.Results Agonist and OGD induced the over-expression of TLR3 and TLR3 mRNA(P0.05).Compared with the cells in the group of OGD,it had better state and more amount in the group which was pretreated with poly(I:C)-LMW(P0.05).Conclusion TLR3 is involved in injured process which OGD induced primary cortical neuron.And injuries of cells are alleviated after pretreated with poly(I:C)-LMW.