【摘要】 目的 探讨氧糖剥夺再灌注(OGD/R)损伤后腺苷酸活化蛋白激酶(AMPK)对小胶质细胞介导的炎性介质释放及信号传导的影响。方法 培养BV2细胞株,应用OGD 6h再灌注24 h建立缺血再灌注损伤离体模型,AICAR(5μmol/L、50μmol/L、100μmol/L)或Compound C(0.1μmol/L、1μmol/L、10μmol/L)不同程度激活或抑制AMPK磷酸化,MTT法检测细胞活性,免疫印迹及ELlSA方法,检测OGD/R后BV2细胞AMPK活性变化,以及时NF-κB信号通路中IκB磷酸化、TNF-α释放的影响。结果 各浓度AICAR均能够促进OGD/R后BV2细胞存活(P<0.05),抑制IκB磷酸化水平(P<0.05),AICAR(100μmol/L)能够增加AMPK磷酸化水平(P<0.05),抑制TNF-α的释放(P<0.01)。而Compound C(10μmol/L)促进BV2细胞死亡(P<0.01),降低OGD/R后AMPK及IκB磷酸化水平(P<0.05),促进TNF-α释放(P<0.05)。结论 AMPK磷酸化激活能够减轻OGD/R后BV2细胞介导的炎性损伤作用,而抑制AMPK磷酸化能够加重神经炎性反应。更多还原

【Abstract】 Objective To study the effects of AMP-activated protein kinase(AMPK) on microglia mediated inflammatory response in oxygen glucose deprivation/reperfusion injury(OGD/R).Methods An in vitro ischemia model was induced by oxygen and glucose deprivation 6 h and reperfusion 24 h in BV2 cell.AMPK was activated and inhibited respectively by AICAR(5 μmol/L,50 μmol/L,100 μmol/L) and Compound C(0.1 μmol/L,1 μmol/L,10 μmol/L) at different concentration.The survival rate in BV2 cell after OGD/R was observed by MTT method.The phosphorylation levels of AMPK and IkB were detected by Western blot.The release of TNF-α was detected by ELISA.Results Various concentrations of AICAR were able to promote BV2 cells survival(P<0.05) and inhibit the phosphorylation level of IκB(P<0.05)in OGD/R.AICAR(100 μmol/L) could increase the phosphorylation level of AMPK(P<0.05),inhibit the release of TNF-α(P<0.01) in OGD/R.While Compound C(10 μmol/L) promoted BV2 cell death(P<0.01),decreased the phosphorylation levels of AMPK and IkB(P<0.05),which could promote the release of TNF-α(P<0.05).Conclusions Activation of AMPK by phosphorylation can relieve the inflammatory damage mediated by OGD/R in BV2 cells,while inhibit the phosphorylation of AMPK can aggravate the inflammatory reaction.更多还原