过刊浏览

血管紧张素II诱导血管外膜成纤维细胞表达炎症介质

陈闻东, 楚玉峰, 李晓东, 高平进

上海交通大学医学院附属瑞金医院，上海市高血压研究所，医学基因组学国家重点实验室，上海 200025；中国科学院上海生命科学研究院健康科学研究所血管生物学实验室，上海 200025；山东大学附属省立医院，济南 250021

摘要

血管外膜成纤维细胞可能是血管炎症反应的重要参与者。本研究旨在探讨血管外膜成纤维细胞在血管紧张素II (angiotensin II, AngII)诱导下能否表达包括趋化因子、黏附分子、炎症因子在内的各种炎症介质及其对炎性细胞的黏附、趋化作用。以AngII诱导的血管外膜成纤维细胞和AngII灌注的大鼠分别进行细胞水平和动物水平的研究。用1 × 10−7 mol/L AngII诱导体外培养的大鼠胸主动脉外膜成纤维细胞，real-time RT-PCR检测各炎症介质mRNA表达，免疫印迹、酶联免疫吸附法(ELISA)检测炎症介质蛋白表达和分泌。使用小鼠RAW264.7单核/巨噬细胞系进行体外细胞黏附和Transwell法体外细胞迁移趋化实验。结果显示，检测的12种炎症介质中，AngII上调血管外膜成纤维细胞中4种炎症介质包括细胞间黏附分子1 (ICAM-1)、P选择素(P-selectin)、单核细胞趋化蛋白1 (MCP-1)、白细胞介素6 (IL-6) mRNA的表达；AngII上调ICAM-1、P-selectin蛋白表达和IL-6分泌，但对MCP-1的分泌量无影响。AngII诱导后的血管外膜成纤维细胞及其上清培养液能增加RAW264.7单核/巨噬细胞的黏附和迁移。采用皮下埋泵输入AngII (200 ng/kg per min)2周的方法制备大鼠整体模型，取大鼠的胸主动脉进行免疫荧光染色。结果显示AngII灌注大鼠的胸主动脉外膜侧有明显CD68阳性细胞聚集，且外膜侧ICAM-1、P-selectin、MCP-1和IL-6的表达都较对照组有不同程度上调。本研究结果表明血管外膜成纤维细胞在AngII诱导下能表达分泌ICAM-1等炎症介质，这些炎症介质可能参与黏附、趋化单核/巨噬细胞，提示血管外膜成纤维细胞可能介导血管的炎症反应。

关键词： 血管外膜成纤维细胞; 血管紧张素II; 炎症; 炎症介质

分类号：R331；Q463

[Angiotensin II induces expression of inflammatory mediators in vascular adventitial fibroblasts.] [Article in Chinese]

Chen Wen-Dong, CHU Yu-Feng, LI Xiao-Dong, GAO Ping-Jin

State Key Laboratory of Medical Genomics and Shanghai Key Laboratory of Vascular Biology at Ruijin Hospital and Shanghai Institute of Hypertension, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China； Laboratory of Vascular Biology, Institute of Health Science, Shanghai Institute for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS), Shanghai 200025, China； Provincial Hospital Affiliated to Shandong University, Jinan 250021, China

Abstract

ascular adventitial fibroblasts (AF) may play an important role in vascular inflammation. This study was aimed to investigate the expression pattern of inflammatory mediators in AF induced by angiotensin II (AngII) and to explore the effects of AF-derived inflammatory mediators on the adhesion and migration of macrophages both in vitro and in vivo. We used real-time RT-PCR to detect the mRNA expression of inflammatory mediators in cultured AF. The results showed that AngII (1 × 10−7 mol/L) up-regulated mRNA expression of 4 inflammatory mediators, including P-selectin, ICAM-1, IL-6 and MCP-1, in cultured AF. Western blot analysis or ELISA revealed that AngII up-regulated P-selectin and ICAM-1 protein expression and IL-6 secretion in cultured AF, but did not alter MCP-1 secretion. We further detected the effects of AF-derived inflammatory mediators on the adhesion and chemotaxis of RAW264.7, a macrophage cell line. We found that AF stimulated with AngII could enhance the adhesion of RAW264.7 and the conditioned medium from AngII-stimulated AF could enhance the migration of RAW264.7. Immunofluorescence study showed an enhanced accumulation of CD68 positive cells and the up-regulation of P-selectin, ICAM-1, IL-6 and MCP-1 in aortic adventitia of AngII-infused (200 ng/kg per min for 2 weeks) rats. We concluded that AF may contribute to vascular inflammation via expression of certain inflammatory mediators and the subsequent adhesion and chemotaxis of macrophages.

Key words: adventitial fibroblast; angiotensin II; inflammation; inflammatory mediators

收稿日期：2015-05-25　　录用日期：2015-09-08

通讯作者：高平进　　E-mail: gaopingjin@sibs.ac.cn

引用本文：

陈闻东, 楚玉峰, 李晓东, 高平进. 血管紧张素II诱导血管外膜成纤维细胞表达炎症介质[J]. 生理学报 2015; 67 (6): 603-610.

Chen Wen-Dong, CHU Yu-Feng, LI Xiao-Dong, GAO Ping-Jin. [Angiotensin II induces expression of inflammatory mediators in vascular adventitial fibroblasts.] [Article in Chinese]. Acta Physiol Sin 2015; 67 (6): 603-610 (in Chinese with English abstract).