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

摘要

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 20μmol · L-1 , respectively) and AGEs (100 mg · L ) , and incubated for 24 h. Meanwhile, a group managed with serum-free culture solution was assumed as blank control. We determined BH4 , NO, 0 " 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.%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.