硝化应激参与衰老血管内皮功能障碍

doi:10.3969/j.issn.1006-7795.2015.06.013

首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (6): 908-914.doi: 10.3969/j.issn.1006-7795.2015.06.013

硝化应激参与衰老血管内皮功能障碍

马语曼, 孙琪, 董玉, 王环愿, 王雯

首都医科大学基础医学院生理学与病理生理学系首都医科大学代谢紊乱相关心血管疾病北京市重点实验室, 北京 100069

收稿日期:2015-06-11 出版日期:2015-12-21 发布日期:2015-12-18
通讯作者: 王雯 E-mail:wangwen@ccmu.edu.cn
基金资助:
国家自然科学基金(81370450)。

Nitrative stress participates in aging-related vascular endothelial dysfunction

Ma Yuman, Sun Qi, Dong Yu, Wang Huanyuan, Wang Wen

Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing Key Laboratory of Metabolic Disorders Related Cardiovascular Diseases, Capital Medical University, Beijing 100069, China

Received:2015-06-11 Online:2015-12-21 Published:2015-12-18
Supported by:
This study was supported by National Natural Science Foundation of China(81370450).

摘要/Abstract

摘要： 目的观察硝化应激对自然衰老大鼠血管内皮形态和功能的影响,探讨衰老时血管内皮功能障碍的机制。方法 3月龄和20月龄两个年龄段健康雄性SD大鼠各12只,分组如下:1 青年对照组(3月龄对照组);2 青年+过氧亚硝基(peroxynitrite,ONOO^-)清除剂内消旋-四(N-甲基-4-吡啶)卟啉五氯化钠(Ⅲ)〔Fe(III) meso-tetra(N-methyl-4-pyridyl) porphine pentachloride,FeTMPyP,3月龄〕;3 老年对照组(20月龄对照组);4 老年+过氧亚硝基清除剂组(FeTMPyP,20月龄)。为抑制过氧亚硝基,腹腔注射给予过氧亚硝基清除剂FeTMPyP(3 mg/kg,每3天1次,共5次)。蛋白印记法(Western blotting)检测肝脏组织衰老相关蛋白P53和P21表达情况;离体胸主动脉血管环灌流,观察血管环对不同浓度乙酰胆碱(acetylcholine,ACh)的反应;HE染色观察胸主动脉血管结构改变;免疫组织化学法检测血管3-硝基酪氨酸(3-nitrotyrosine,3-NT)、内皮标志物血管假性血友病因子(von willebrand factor,vWF)及内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)。结果 20月龄衰老大鼠胸主动脉内皮依赖性舒张功能较3月龄大鼠显著下降,采用过氧亚硝基的清除剂FeTMPyP干预可明显提高衰老血管内皮依赖性舒张功能;HE及免疫组织化学染色结果显示20月龄衰老大鼠胸主动脉较3月龄大鼠均出现较明显的损伤,采用过氧亚硝基的清除剂FeTMPyP干预可明显减轻衰老血管形态学损伤;免疫组织化学结果显示衰老大鼠胸主动脉3-NT升高,提示出现明显的硝化应激反应,采用过氧亚硝基的清除剂FeTMPyP干预可明显降低3-NT。结论衰老时过氧亚硝基生成增多,硝化应激反应增强,参与血管形态学损伤和血管内皮依赖性舒张功能降低。

关键词: 衰老, 硝化应激, 血管内皮

Abstract: Objective This study aimed to observe the effect of nitrative stress on the morphology and function of vascular endothelium in the natural aging rats. Methods Young(3 months) and aging(20 months) male SD rats(SPF grade) were randomly divided into four groups, with ten rats in each group: 1 3 months control;2 3 months+Fe(III) meso-tetra(N-methyl-4-pyridyl) porphine pentachloride(FeTMPyP);3 20 months control;4 20 months+FeTMPyP. The experimental groups were given FeTMPyP(peroxynitrite scavenger;3 mg/kg every 3 days;totally 5 times) via intraperitoneal injection. The levels of aging-related proteins P53 and P21 in liver were detected by Western blotting. Rat thoracic aortas was isolated and connected to an isometric force transducer. Endothelium-dependent dilation was measured by classic drugs, such as different concentrations of acetylcholine(ACh). The histological changes of thoracic aortas were observed by HE staining. The 3-nitrotyrosine and endothelium markers vWF and endothelial nitric oxide synthase(eNOS) levels in vascular tissues were detected by immunohistochemistry. Results In 20 months old rats, the endothelium-dependent relaxation in aortas was dropped significantly than the 3 months rats. Adoption of peroxynitrite scavenger FeTMPyP intervention can obviously improve the endothelium-dependent relaxation of aging rats. Both HE and von willebrand factor(vWF) & eNOS immunohistochemistry staining results showed that there were obvious morphological damage in thoracic aorta in 20 months old rats. Adoption of peroxynitrite scavenger FeTMPyP intervention can obviously reduce the damage of aging vascular morphology. Immunohistochemistry results showed that in aged thoracic aortas, the 3-nitrotyrosine levels increased significantly, which indicated the obvious nitrative stress reaction. Adoption of peroxynitrite scavenger FeTMPyP intervention can obviously reduce the 3-nitrotyrosine levels. Conclusion During aging, the increased formation of peroxynitrite and enhanced nitrative stress reaction can lead to endothelial injury and functional decline.

Key words: aging, nitrative stress, vascular endothelium

中图分类号:

R363

马语曼, 孙琪, 董玉, 王环愿, 王雯. 硝化应激参与衰老血管内皮功能障碍[J]. 首都医科大学学报, 2015, 36(6): 908-914.

Ma Yuman, Sun Qi, Dong Yu, Wang Huanyuan, Wang Wen. Nitrative stress participates in aging-related vascular endothelial dysfunction[J]. Journal of Capital Medical University, 2015, 36(6): 908-914.

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硝化应激参与衰老血管内皮功能障碍

Nitrative stress participates in aging-related vascular endothelial dysfunction

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