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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5,10,20μmol·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。
Objective To observe the effect of probucol on endothelial nitric oxide synthase (eNOS) decoupling in cardiac microvascular endothelial cells and to explore its mechanism. Methods Cardiac microvascular endothelial cells were treated with 100 mg · L-1 bovine serum albumin (BSA-AGEs) and 5, 10, 20 μmol·L-1 probucol for 24 h, and the concentrations of tetrahydrobiopterin (BH4), nitric oxide (NO) and superoxide anion (O2-). The expression of eNOS protein was detected by immunohistochemical staining. ROS was detected by fluorescent staining and p47phox protein was detected by Western blot. Results With the increase of probucol concentration, NO production increased, O2- production decreased, eNOS expression decreased, BH4 content increased, ROS expression decreased, p47phox expression decreased (P <0.01 or P <0.05). Conclusion Probucol can inhibit AGEs-induced eNOS decoupling in cardiac microvascular endothelial cells, which may be related to the inhibition of NADPH oxidative stress.