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目的:探讨丹皮酚(Paeonol,Pae)对血管紧张素II(AngiotensinⅡ,AngⅡ)所致动脉内皮功能障碍的保护作用及其机制。方法:采用实时定量聚合酶链式反应(RT-PCR)和Westernblot检测酸性鞘磷脂酶(acidsphingomyelinase,ASM)基因及蛋白表达;采用免疫组织荧光检测血管环神经酰胺(ceramide,Cer)含量变化;采用血管环张力描记技术检测主动脉血管环的舒张功能;采用二氢乙啶荧光探针(dihydroethidium,DHE)检测血管环超氧阴离子(superoxideanion,O2-·)水平。结果:与对照组相比,AngⅡ孵育后主动脉对乙酰胆碱(acetylcholine,Ach)的舒张反应显著降低,Pae与AngⅡ共孵育则无明显改变;AngⅡ及Pae孵育后,动脉对硝普钠(sodiumnitroprusside,SNP)的舒张反应均无明显变化;AngⅡ孵育可增高动脉环O2-·水平,这一反应被Pae明显抑制;与对照组相比,AngⅡ孵育动脉ASMmRNA水平无明显改变,但蛋白表达显著增加,Cer含量显著增多,Pae与AngⅡ共孵育动脉ASM蛋白及Cer水平与对照组相比均无明显改变;采用神经酰胺酶抑制剂N-oleoylethanolamine(OEA)增加动脉Cer含量后,Pae降低动脉O2-·水平以及增强Ach舒张反应的效应被显著抑制。结论:Pae通过ASM/Cer通路改善AngⅡ所致动脉氧化应激增强和内皮功能障碍。
Objective: To investigate the protective effect of Paeonol on arterial endothelial dysfunction induced by Angiotensin Ⅱ (AngⅡ) and its mechanism. Methods: The gene and protein expression of acidphingomyelinase (ASM) were detected by real-time quantitative polymerase chain reaction (RT-PCR) and Western blotting. The content of ceramide (Cer) in vascular ring was detected by immunohistochemistry. Vascular tension-ring technique was used to detect the diastolic function of aortic rings. The level of superoxide anion (O2-) was detected by dihydroethidium (DHE). Results: Compared with the control group, the relaxation of achilles to acetylcholine (Ach) in the aorta was significantly decreased after AngⅡ incubation and no significant change was found in the co-incubation of Pae with AngⅡ. After the incubation with AngⅡ and Pae, the aortas were exposed to sodiumnitroprusside SNP). No significant changes were found in the diastolic response of AngⅡmice. AngⅡ incubation increased the level of O2- in arterial rings, which was significantly inhibited by Pae. Compared with the control group, there was no significant change in ASMmRNA levels in AngⅡmice but the protein expression was significantly increased, Cer significantly increased, Pae and Ang Ⅱ co-incubated ASM protein and Cerium levels compared with the control group had no significant change; the ceramide inhibitor N-oleoylethanolamine (OEA) increased Cerium content, Pae decreased arterial O2- · The level and the effect of enhancing Ach relaxation were significantly suppressed. Conclusion: Pae can improve the arterial oxidative stress induced by Ang Ⅱ and endothelial dysfunction through ASM / Cer pathway.