晚期糖基化终产物促进血管外膜成纤维细胞迁移机制及坎地沙坦的干预作用研究  

Mechanism of migration of vascular adventitial fibroblasts enhanced by advanced glycation end-products and the inhibition effect of candesartan

作　　者：刘亚洋[1] 梁春[1] 吴宗贵[1] 

机构地区：[1]第二军医大学长征医院心内科,上海200003

出　　处：《解放军医学杂志》2009年第5期523-527,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家重点基础研究发展计划(973计划)资助项目(2005CB523309)

摘　　要：目的探讨晚期糖基化终产物(AGEs)对血管外膜成纤维细胞(AF)迁移的影响及坎地沙坦的干预作用。方法组织贴块法培养SD大鼠的AF细胞,用Transwell小室检测AGEs对AF迁移的影响,RT-PCR及免疫印迹技术观察AGEs对AF晚期糖基化终产物受体(RAGE)及丝裂原活化蛋白激酶(MAPK)通路磷酸化的影响。结果不同浓度的AGE(50、100、150、200、300mg/L)均可从基因和蛋白水平上调RAGE的表达,且均在200mg/L时达到峰值(P<0.05)。p38拮抗剂、ERK1/2拮抗剂、JNK拮抗剂、坎地沙坦可以抑制由糖基化人血清白蛋白(AGE-HAS)刺激引起的RAGE表达(P<0.05)。AGEs可促进p38、ERK1/2、JNK的磷酸化,JNK在20min,p38、ERK1/2在30min时磷酸化达峰值(P<0.05)。p38拮抗剂、ERK1/2拮抗剂、JNK拮抗剂可分别抑制p38、ERK1/2、JNK磷酸化,RAGE中和抗体、坎地沙坦可抑制p38、ERK1/2、JNK磷酸化。不同浓度AGE(50、100、150、200、300mg/L)可促进AF细胞的迁移,该作用于200mg/L时达到峰值(P<0.05)。RAGE中和抗体、p38拮抗剂、ERK1/2拮抗剂、坎地沙坦均可抑制AGE所致成纤维细胞的迁移(P<0.01)。结论AGEs经由RAGE影响MAPK通路而促进AF细胞迁移,坎地沙坦可通过阻断此途径抑制AF细胞迁移,这可能是其血管保护作用的机制之一。Objective To investigate the mechanism of migration of vascular adventitial fibroblasts （AFs） enhanced by advanced glycation end-products （AGEs） and the inhibition effect of eandesartan. Methods Isolated vascular adventitial fibroblast of Sprague-Dawley （SD） rats were cultured. Migratory potential was estimated with Transwell chamber in vitro. Expression of receptor for AGEs （RAGE） and phosphorylated mitogen activated protein kinase （MAPK） of AFs were determined by RT-PCR and Western blotting. Results . AGEs modified human serum albumin （AGE-HAS, concentration ranged from 0 to 300mg/L） up-regulated the expression of RAGE at mRNA and protein levels, peaked at a concentration of 200mg/L （P〈0. 05）. This effect could be significantly inhibited by p38, ERK1/2 and JNK MAPK inhibitors as well as candesartan （P〈0. 05）. AGEs increased phosphorylation of p38, ERK1/2 and JNK MAPK in AFs in a timedependent manner. JNK peaked at 20rnin and p38 ERK1/2 peaked at 30min （P〈0. 05）. Phosphorylation of p38, ERK1/2 and JNK was suppressed by p38, ERK1/2 and JNK MAPK inhibitor, respectively. This effect could also be significantly inhibited by candesartan and anti-RAGE neutralizing antibody. The migration of AFs enhanced by AGEs （concentration ranged from 0 to 300mg/L） was markedly increased in a dose-dependent manner, peaked at a concentration of 200mg/L （P〈：0. 05）. Anti-RAGE neutralizing antibody, p38, ERK1/2 MAPK inhibitors, as well as candesartan suppressed AGEs-induced migration of AFs （ P〈0. 01）. Conclusions AGEs promote AFs migration via RAGE and MAPK pathway. Candesartan can effectively inhibit the migration of AFs. It might be a novel mechanism of vascular protective effect of candesartan.

关 键 词：糖基化终产物 高级 成纤维细胞 细胞运动 丝裂原激活蛋白激酶类 坎地沙坦 

分 类 号：R341.84[医药卫生—基础医学]