丹参酚酸B通过激活过氧化体增殖物激活型受体γ抑制树突状细胞免疫成熟的作用及其机制  

Effect of ERK1/2 Signal Transduction Pathway in Vascular Endothelial Cell Apoptosis

作　　者：刘红樱[1] 孙爱军[1,2] 王时俊[1,2] 史大卓[3] 徐磊[1] 程勇[1] 邹云增[1,2] 王克强[1] 陈可冀[3] 葛均波[1,2] 

机构地区：[1]复旦大学附属中山医院上海市心血管病研究所,上海市200032 [2]复旦大学生命科学院,上海市200433 [3]中国中医科学院附属西苑医院,北京市100091

出　　处：《中国动脉硬化杂志》2011年第3期265-265,共1页Chinese Journal of Arteriosclerosis

摘　　要：研究背景树突状细胞(DCs)是体内功能最强大的抗原递呈细胞,具有独特的激活初始T淋巴细胞的功能,是启动和调控特异性免疫应答的中心环节。近年来的大量基础和临床研究证实,动脉粥样硬化(As)是一种慢性炎症和免疫性疾病,DCs直接或间接参与As的发生发展。以DCs为作用靶点可能是干预As的有效方法。丹参酚酸B(Sal B)是自中药丹参中提取出的水溶性单体,是丹参的重要药理活性成份,化学结构明确,性质稳定。以往的研究证实,Sal B对心、脑、肝、肾等多个器官具有重要的保护作用。近期的研究还发现,Sal B具有抗炎症、抗免疫反应的作用,能够影响As的发生发展,但具体作用机制和靶点还不明确。目的从免疫炎症的角度探讨Sal B对氧化低密度脂蛋白(ox-LDL)诱导的人单核细胞源DCs免疫功能成熟的影响,进一步研究其作用机制,为临床As的防治提供新靶点和新思路。方法培养人单核细胞源DCs,Sal B预处理后,再与ox-LDL共孵育。流式细胞术检测DCs表面分子(CD40、CD1a、CD86和HLA-DR)的表达,ELISA法检测细胞培养上清液细胞因子(IL-12和TNF-α)的浓度,Western blot法检测PPARγ和MAPKs的蛋白表达,RT-PCR法检测PPARγ的基因表达,Luciferase报告系统检测PPARγ的DNA结合活性。结果与ox-LDL组相比,Sal B预处理组DCs的表面分子CD40、CD1a、CD86和HLA-DR的表达明显下降(P<0.05),细胞因子IL-12和TNF-α的浓度明显降低(P<0.01),证明Sal B预处理明显抑制ox-LDL诱导的人单核细胞源DCs的免疫功能成熟。与ox-LDL组相比,Sal B预处理组DCs的MAPKs的蛋白表达明显下调,主要是P38蛋白表达明显降低(P<0.05),表明Sal B预处理明显抑制ox-LDL诱导的人单核细胞源DCs的MAPKs蛋白表达。进一步采用siRNA技术使PPARγ基因沉默后,Sal B预处理抑制ox-LDL诱导的人单核细胞源DCs表面分子、细胞因子和MAPKs蛋白表达的作用被明显翻转,提示Sal B通过影响PPARγ抑�Aim To explore the changes in extracellar signal-regulated prote in kinase（ERK1 /2） in endothelial cell induced by Angiotensin Ⅱ at the different time courses,and its possible molecular mechanism.Method s Human umbilicalve in endothe-lial cell（HUVEC） were cultured in vitro and intervened by AngⅡ.HUVEC were divided into 2 groups,the control group,AngⅡ group（stimulated by AngⅡ10-6mol/L for 24h）.Flow cytometery with Annexin V-FITC /PI double staining and Hoechst33258 fluo-rescence staining were used to detect apoptosis of HUVEC.The expressions of apoptosis-association genes Bcl-2,Bax were detected by RT-PCR and ERK1 /2 levels were detected by Western-blotting at different time points.Results 10-6mol/L Angiotensin Ⅱ stimulation stimulated cell apoptosis.Bcl-2mRNA levels were time-dependently decreased,the radio of Bcl-2 /Bax was decreased markedly（P0.05）.Phosphorylation of ERK1 /2 began to increase and reach the peak at 18 h（P0.01）.Conclusions Cell apoptosis is possibly important factor for atherosc lerosis.One of its molecular mechanisms might be associated with decreasing the expression level of Bcl-2 and the radio of Bcl-2 /Bax.There is a probability that activated ERK1/2 signal pathway is involved in the process of pathologic and physiologic reaction in the apoptosis of endothelial cell induced by Angiotensin Ⅱ.

关 键 词：丹参酚酸B 过氧化体增殖物激活型受体Γ 树突状细胞 氧化低密度脂蛋白 动脉粥样硬化 

分 类 号：R363[医药卫生—病理学]