血管紧张素Ⅱ对单核细胞组织因子表达的影响及调控机制  

Effect of angiotensin Ⅱ on tissue factor expression in human peripheral blood monocytes and its mechanisms

作　　者：何美霞[1] 何晓凡[1] 解勤之[2] 陈方平[2] 贺石林[1] 

机构地区：[1]中南大学湘雅医学院生理教研室 [2]中南大学湘雅医院血液科

出　　处：《中华血液学杂志》2003年第9期470-473,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金重点资助项目 (3 983 0 180 )

摘　　要：目的　观察血管紧张素Ⅱ (AngⅡ )对人单核细胞组织因子 (TF)表达的影响及机制。方法　人外周血单核细胞的分离采用淋巴细胞分离液及Percoll。细胞促凝活性 (PCA)的检测采用一期凝固法。细胞TF抗原测定采用ELISA法 ;TFmRNA检测采用RT PCR的方法 ;IκBα水平分析采用Westernblot方法 ;NF κB的变化分析用凝胶电泳迁移率实验。结果　AngⅡ (10 - 9～ 10 - 7mol L)可剂量依赖性诱导单核细胞PCA、TF抗原及mRNA的增加 ,并有明显的时效关系。洛沙坦 (losartan)浓度在 10 - 6 ～10 - 5mol L可不同程度地阻断AngⅡ的作用。Staurosporine(2 .5× 10 - 7mol L)以及金雀异黄素 (4× 10 - 5mol L)可显著抑制AngⅡ (10 - 7mol L)诱导的单核细胞PCA、TF抗原增加及TFmRNA的表达 ;AngⅡ(10 - 7mol L)作用 15min可引起单核细胞IκBα水平下降 (P <0 .0 5 ) ,6 0min时IκBα水平降至最低水平 ,180min时恢复至正常水平 ;凝胶电泳迁移率显示AngⅡ (10 - 7mol L)诱导单核细胞NF κB的作用在 15min起效 ,6 0min时核内NF κB的水平最高 ,180min恢复正常 ;洛沙坦 (10 - 5mol L)或PDTC(10 - 4mol L)均可抑制NF κB的活化。结论　AngⅡ可诱导单核细胞TF的表达 ,该作用是通过AngⅡ 1型受体实现的。胞内PKC发挥较强的作用。NF κB的?Objective To elucidate the effect of angiotensin Ⅱ(Ang Ⅱ) on the expression of tissue factor (TF) by monocytes and its mechanisms. Methods Monocytes were isolated from healthy volunteers by Ficoll-Hypaque gradient and Percoll, and cultured in RPMI-1640. Procoagulant activity (PCA) was determined by one-stage clotting method, TF antigen by ELISA. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the TF gene mRNA. The levels of IκBɑ was detected by Western blot. Electrophoretic mobility shift assays (EMSA) were performed to evaluate the activity of NF-κB. Results AngⅡ(10 -9 ～10 -7 mol/L) significantly increased monocyte PCA,TF antigen and TF mRNA expression in a dose and time dependent manner. Losartan (10 -6 ～10 -5 mol/L) significantly inhibited the effects of Ang Ⅱon TF activity, antigen and mRNA expression in a dose-dependent effects. Staurosporine (2.5×10 -7 mol/L)and genistein(4×10 -5 mol/L) lowered TF level of monocytes (P<0.05). Western blot analysis revealed that after exposure to AngⅡ(10 -7 mol/L), IκBα level decreased at 15 min, reached nadir at 60 min, and recovered at 180 min. EMSA showed NF-κB binding activity increased at 15 min, reached peak at 60 min, and recovered at 180 min. Pyrrolidine dithiocarbamate (PDTC,10 -4 mol/L), an inhibitor of NF-κB, or AT1R antagonist losartan(10 -5 mol/L)inhibited AngⅡ-induced NF-κB translocation. Conclusions Ang Ⅱ could induce the expression of TF in human monocytes, and this effect was mediated by AT1R. The PKC pathway played the most important role in AngⅡ-induced TF expression. The activation of NF-κB was involved in TF expression in monocytes.

关 键 词：血管紧张素Ⅱ 单核细胞组织因子 核转录因子-ΚB TF抗原 

分 类 号：R363[医药卫生—病理学]