地氟醚预处理对缺氧/复氧损伤内皮细胞内游离Ca~(2+)浓度和钙网蛋白表达的影响

摘要

目的 观察地氟醚预处理对缺氧/复氧(A/R)损伤的内皮细胞内游离Ca~(2+)浓度及钙网蛋白表达水平的影响.方法 人脐静脉内皮细胞株(ECV304)细胞分为三组:A/R组(Ⅰ组)、地氟醚1.0 MAC预处理+A/R组(Ⅱ组)和空白对照组(Ⅲ组).应用Fluo-3/AM荧光探针法和Western blot方法检测细胞中游离Ca~(2+)浓度和钙网蛋白水平.结果 与Ⅲ组比较,Ⅰ和Ⅱ组细胞中游离Ca~(2+)浓度显著增高(P<0.01),但Ⅱ组的升高幅度明显低于Ⅰ组(P<0.05);Ⅱ组细胞中游离Ca~(2+)负载有所增高,但钙网蛋白表达水平增加(P<0.05).结论 地氟醚预处理可通过降低细胞内的钙超载来减轻A/R造成的损伤.%Objective To observe the effects of desflurane preconditioning on the intracellular free Ca~(2+) and calreticulin expression in the endothelial cells Ⅰ underwent anoxia／reoxygenation (A／R).Methods Human umbilical vein endothelial cells(ECV304)were divided into three groups of A／R(group Ⅰ),Des + A／R(group Ⅱ)and control(groupⅢ).The concentrations of free Ca~(2+) were measured with Fluo-3／AM florescent probe and the expressions of calreticulin(CRT)protein were detected with Western blot.Results Compared with group Ⅲ,the concentration of free Ca~(2+) was higher and the expression of CRT was lower in group Ⅰ(P<0.01),which were opposite in group Ⅱ (P<0.05).Conclusion Desflurane preconditioning can protect the endothelial cells by attenuating the Ca~(2+) overload caused by A／R injury.