地氟醚预处理对缺氧/复氧损伤ECV304细胞NF-κB活性的影响

The in vitro effect of desflurane preconditioning on nuclear factor-κB activation induced by anoxia /reoxygenation injury in ECV304 cells

目的探讨地氟醚预处理对缺氧/复氧(A/R)损伤ECV304细胞NF-κB活性的影响。方法选用ECV304细胞株.实验分为两部分:(1)将细胞分为空白对照组及TNF-α不同持续时间刺激组共6组,用West-ernblot法分析NF-κB的抑制因子IκB蛋白的降解和磷酸化。(2)将细胞分为5组,即空白对照组(Ⅰ组)、A/R组(Ⅱ组)、A/R+TNF-α10ng/mL刺激组(Ⅲ组)、地氟醚1.0MAC预处理+A/R组(Ⅳ组)和地氟醚1.0MAC预处理+A/R+TNF-α10ng/mL刺激组(Ⅴ组)。用间接免疫荧光法检测各组细胞NF-κB/p65亚基的定位。结果TNF-α刺激ECV304细胞后,IκB-α蛋白降解的时间高峰为30min,而IκB-α磷酸化的时间高峰为1h。A/R损伤可激活ECV304细胞中NF-κB的活性,而经1.0MAC地氟醚预处理后,由TNF-α激活的上述变化可受到抑制。免疫荧光分析显示,TNF-α刺激可使NF-κB/p65荧光标记由胞浆区转入胞核区,而经地氟醚预处理后,NF-κB/p65入核明显减少。结论抑制NF-κB的激活可能是地氟醚预处理减少ECV304细胞缺氧/复氧损伤的机制之一。

Purpose To investigate the effect of desflurane preconditioning on nuclear factor-κB(NF-κB) activation induced by anoxia /reoxygenation(A/R) injury in ECV304 cells in vitro.Methods Cell line was used.In Part A experiment,the ECV304 cells were divided into 6 groups:the control group and the other five groups stimulated by TNF-α with different duration.The protein levels of IκB-α(the inhibitory factor of the NF-κB) and its phosphorylation form,p-IκB-α,were detected by Western blot method.In Part B experiment,...