摘要
目的:研究炎症状态下血管平滑肌细胞中沉默调节蛋白1(SIRT1)的表达及其相关的信号通路。方法:以大鼠左颈总动脉球囊损伤和原代血管平滑肌细胞为模型,采用免疫组化的方法研究颈总动脉球囊损伤后SIRT1蛋白表达;采用免疫荧光观察血管平滑肌中TNF-α对SIRT1表达和定位的影响;采用Western blotting检测TNF-α对血管平滑肌细胞中SIRT1表达及p42/p44促分裂素原活化蛋白激酶(MAPK)表达的影响;并考察用MAPKs抑制剂处理后TNF-α对SIRT1表达的影响。结果:免疫组化结果显示球囊损伤的大鼠颈总动脉平滑肌细胞中SIRT1表达增加;免疫荧光显示SIRT1主要定位于血管平滑肌细胞核且TNF-α处理后SIRT1表达增加;Western blotting结果表明TNF-α处理的大鼠血管平滑肌细胞中SIRT1和磷酸化p42/p44 MAPK表达高于对照组;用p42/p44 MAPK抑制剂处理后,TNF-α诱导的SIRT1表达被抑制。结论:球囊损伤及TNF-α处理均可增高血管平滑肌细胞中SIRT1表达,p42/p44 MAPK信号通路参与对这一过程的调控。
Objective： To investigate the sirtuin 1（ SIRT1） expression regulation in vascular smooth muscle cells under inflammatory injury and the related signal pathway involved in the process. Methods： Balloon injury animal model and primary vascular smooth muscle cells（ VSMCs） were used in this study. Immunohistochemistry was used to detect SIRT1 expression in injured carotid artery. SIRT1 expression and localization were detected by immunofluorescence. Western blotting was used to detect the effect of TNF-α on SIRT1 or phospho-p42/p44 MAPK expression in VSMCs with or without the treatment of MAPKs pathway inhibitors. Results： Immunohistochemistry result showed that SIRT1 protein expression was enhanced in injured carotid artery. Immunofluorescence images showed that SIRT1 was mainly localized in nucleus in VSMCs and the expression of SIRT1 was triggered by TNF-α. Consistently,Western blotting showed that SIRT1 and phospho-p42/p44 MAPK was increased in TNF-α-treated VSMCs. More importantly,the inhibitor of p42/p44 MAPK pathway suppressed the elevated-SIRT1 by TNF-α in VSMCs. Conclusion： SIRT1 protein level was increased in VSMCs exposed to balloon injury or TNF-α treatment,and p42/p44 MAPK pathway was involved in the process of TNF-α-induced SIRT1 expression.
作者
张慧娜
李玉
彭露
杨云云
ZHANG Huina;LI Yu;PENG Lu;YANG Yunyan(Key Laboratory of Upper Airway Dysfunction-related Cardiovascular Diseases,Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)
出处
《心肺血管病杂志》
2018年第7期682-686,共5页
Journal of Cardiovascular and Pulmonary Diseases
基金
国家自然基金应急项目(31741064)
国家自然基金面上项目(81471082)
