白三烯B4对巨噬细胞小凹蛋白-1表达的影响及其与冠状动脉斑块不稳定的关系  

The role of leukotriene B4 on the expression of caveolin-1 in macrophage and its potential association with plaque unstable

作　　者：王琳[1] 江建军[1] 许莎莎[1] 方崇峰[1] 陈葆国[2] 朱敏[2] 黄维清[3] 唐礼江[1] 

机构地区：[1]温州医学院附属台州医院心内科,浙江317000 [2]温州医学院附属台州医院中心实验室,浙江317000 [3]山东省青岛市市立医院东院病理科

出　　处：《中国介入心脏病学杂志》2010年第4期213-218,共6页Chinese Journal of Interventional Cardiology

基　　金：浙江省科技厅基金资助(2006c33019)

摘　　要：目的研究小凹蛋白-1(Cav-1)及白三烯B4(LTB4)与冠状动脉斑块不稳定的关系,观察LTB4及其抑制剂对巨噬细胞Cav-1表达的影响并探讨其意义。方法免疫组化检测41例人尸检冠状动脉粥样斑块中巨噬细胞Cav-1蛋白的表达;以流式细胞技术检测167例入选患者外周血单核细胞Cav-1的表达水平,用ELISA检测血清中LB4的表达水平;分别用浓度为10-8mol/L、10-7mol/L的LTB4及10-7mol/LU75302处理THP-1源性的巨噬细胞,RT-PCR检测细胞内的TNF-αmRNA,Cav-1mRNA水平及Western Blot检测Cav-1蛋白表达的变化。结果与稳定斑块相比,不稳定斑块中巨噬细胞聚集处Cav-1表达下调或表达阴性。急性冠脉综合征(ACS)组中急性心梗(AMI)组及不稳定性心绞痛(UAP)组外周血单核细胞Cav-1表达水平皆显著低于稳定心绞痛(SAP)组、非冠心病组(99.2±40.8MnX比(176.1±90.3)MnX、(246.0±122.8)MnX(t=-4.786、5.112),(104.8±37.4)MnX比(176.1±90.3)MnX、(246.0±122.8)MnX(t=-4.508、-4.942,P<0.01);AMI组血清LTB4表达显著高于UAP组、SAP组及非冠心病组(209.8ng/L比40.7ng/L、59.5ng/L、63.8ng/L,Z=-3.771、-3.426、-2.849,P<0.01)。LTB4可下调Cav-1mRNA及蛋白表达水平(0.963、0.959比1.186,t=-5.079、-5.148),0.712比0.840(t=-4.050,P<0.01),上调肿瘤坏死因子-α(TNF-α)mRNA的表达(0.637比0.555)(t=5.148,P<0.01);U75302预处理组Cav-1mRNA及蛋白水平上调(1.089比0.959、0.963(t=3.018、3.087,0.799比0.712,t=2.849,P<0.05),TNF-αmRNA水平下降(0.529比0.637,t=-3.087,P<0.01)。结论冠状动脉不稳定斑块中巨噬细胞及ACS患者外周血单核细胞Cav-1表达降低,AMI患者血清中LTB4表达上调,提示高水平LTB4及低表达Cav-1与斑块不稳定有关。LTB4下调THP-1来源的巨噬细胞Cav-1mRNA及蛋白的表达,上调TNF-αmRNA水平,U75302预处理可拮抗上述LTB4的作用,提示使用拮抗LTB4的药物可能有稳定粥样斑块的作用。Objective To study the effects of Caveolin-1（Cav-1）and Leukotriene B4（LTB4）on plaque destabilization,and to investigate the effects of LTB4 and LTB4 antagonist on the expression of Caveolin-1 in macrophage and its application.Methods The Cav-1 expression of macrophages in the coronary artery atherosclerotic plaques studied by immunohistochemisty was done in 41 cases of voluntary autopsy,Cav-1 expression on peripheral blood mononuclear cells measured by flow cytometry and serum LTB4 level by ELISA were done in 167 patients.Moreover,the macrophage differentiated from THP-1 was stimulated with LTB4 of different concentrations as 10 -8mol/L,10 -7mol/L and 10 -7mol/L of U75302,RT- PCR was performed to measure the mRNA level of TNF-α and Cav-1.The protein levels of Cav-1 was evaluated by Western Blot.Results Cav-1 expressions were lower or negative in macrophages from unstable plaques than stable ones.Compared with patients with Stable Angina Pectoris（SAP）or without coronary artery disease,the expression of Cav-1 in ACS patients was significantly lower（99.2 ± 40.8 MnX vs 176.1 ± 90.3 MnX and 246.0 ± 122.8 MnX（t = - 4.786 and 5.112）,104.8 ± 37.4 MnX vs 176.1 ± 90.3 MnX and 246.0 ± 122.8 MnX（t = - 4.508 and - 4.942,P 0.01）and the level of LTB4 in AMI patients was higher than the other patients or cases（209.8 ng/L vs 40.7 ng/L and 59.5 ng/L and 63.8 ng/L（Z = - 3.771 and - 3.426 and - 2.849）,P 0.01）.Furthermore,LTB4 down-regulated the expression of Cav-1 mRNA and protein translation in macrophages（0.963 and 0.959 vs 1.186（t = - 5.079 and - 5.148）,0.712 vs 0.840（t = - 4.050）,P 0.01）,but up-regulated the transcription of Tumor necrosis factor-alpha（TNF-α）mRNA（0.637 vs 0.555（t = 5.148）,P 0.01）.Moreover,preincubated with U75302 increased Cav-1 mRNA transcription or Protein translation（1.089 vs 0.959 and 0.963（t = 3.018 and 3.087）,0.799 vs 0.712（t = 2.849）,P 0.05）,but the transcription of TNF-α mRNA was decreased（0.529 vs 0.637,（t = - 3.087）,P 0.01

关 键 词：窖蛋白1 白三烯B4 炎症 急性冠脉综合征 外周血单核巨噬细胞系统 

分 类 号：R363[医药卫生—病理学]