普罗布考对心脏微血管内皮细胞eNOS脱耦联与NADPH氧化应激途径的影响

目的观察普罗布考对心脏微血管内皮细胞内内皮型一氧化氮合酶（eNOS）脱耦联的作用，探讨其作用机制。方法100 mg•L－1牛血清清蛋白糖基化终末产物（BSA AGEs）与5，10，20 μmol•L－1普罗布考作用于心脏微血管内皮细胞24 h，检测四氢生物喋呤（BH4）、一氧化氮（NO）和超氧阴离子（O2 ），免疫组织化学检测eNOS蛋白表达情况，荧光染色检测活性氧簇（ROS），Western blot检测p47phox蛋白。结果随着普罗布考浓度增加，NO生成增加，O2 生成减少，eNOS表达减少，BH4含量增加，ROS表达降低，p47phox表达减少（P＜0.01或P＜0.05）。结论普罗布考能抑制AGEs诱导的心脏微血管内皮细胞eNOS脱耦联，其机制可能与抑制NADPH氧化应激有关。

Effects of Probucol on AGEs-induced eNOS Uncoupling and NADPH Oxidative Stress Pathway

Objective To observe the effect of probucol on AGEs-induced eNOS uncoupling in cardiac microvascular endothelial cells,and to investigate its mechanism.Methods The cardiac microvascular endothelial cells were cultured in vitro.The cells were added with probucol(5,10 and 20 μmol·L-1,respectively) and AGEs(100 mg·L-1),and incubated for 24 h.Meanwhile,a group managed with serum-free culture solution was assumed as blank control.We determined BH4,NO,O2-generation,eNOS,ROS and p47phox protein expression of all groups.Results As probucol concentration was increased,production of NO was increased gradually(P<0.05),production of O2-decreased gradually(P<0.05),BH4 expression increased(P<0.05),eNOS expression gradually decreased(P<0.05),ROS was seen decreased gradually(P<0.05).Conclusion Probucol can inhibit intracellular eNOS uncoupling in the cardiac microvascular endothelial cells,and its mechanism might be related to suppressing the activation of NADPH oxidative stress pathway.