人脐静脉内皮细胞损伤过程中核转录因子κB及单核细胞趋化蛋白1mRNA的变化  

Changes in nuclear factor kappa B and monocyte chemotactic protein 1 mRNA during the process of human umbilical vein endothelial cells injury

作　　者：刘镘利[1] 王宗仁[1] 张筠[1] 雷战军[2] 

机构地区：[1]解放军第四军医大学西京医院中医科,陕西省西安市710032 [2]解放军第四军医大学唐都医院烧伤整形科,陕西省西安市710038

出　　处：《中国组织工程研究与临床康复》2008年第20期3841-3844,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:既往实验表明,炎性因子与内皮细胞的功能有密切的关系,观察保护和损伤因素对体外培养人血管内皮细胞炎性因子作用的报道较少。目的:实验拟观察姜黄素和肿瘤坏死因子α对人脐静脉内皮细胞分泌核因子κB以及单核细胞趋化蛋白1mRNA的影响。设计、时间及地点:细胞观察实验,于2006-12/2007-08在解放军第四军医大学组织工程实验室完成。材料:健康人脐带由解放军第四军医大学唐都医院提供。内皮细胞培养采用M199完全培养基。方法:实验分为3组,正常对照组为无血清M199培养基;肿瘤坏死因子α组:在正常细胞培养液中加入浓度为10μg/L的肿瘤坏死因子α诱导损伤1.5h;姜黄素组:先在培养液中加入10μg/L的肿瘤坏死因子α30min诱导损伤,再分别加入终浓度为6.25,12.5,25,50,100mg/L的姜黄素孵育1h。主要观察指标:①四甲基偶氮唑盐法检测不同浓度姜黄素对人脐静脉内皮细胞增殖的影响。②免疫细胞化学染色法观察胞核内核转录因子κB的表达。③反转录-聚合酶链反应检测单核细胞趋化蛋白1mRNA的表达。结果:姜黄素可以提高肿瘤坏死因子α诱导损伤的人脐静脉内皮细胞活性(P<0.01),并呈一定剂量依赖性。肿瘤坏死因子α可刺激核因子κBp65表达的强度,姜黄素可以减少核因子κBp65的表达。与正常对照组相比,肿瘤坏死因子α作用30min后,单核细胞趋化蛋白1mRNA相对表达量明显增高(P<0.01);加入不同浓度的姜黄素作用1h后,单核细胞趋化蛋白1mRNA相对表达量明显减低(P<0.05,P<0.01)。结论:姜黄素对损伤的内皮细胞具有保护作用,其作用途径可能通过抑制核转录因子κB活性,并下调单核细胞趋化蛋白1mRNA的表达,从而阻滞了单核细胞的聚集。BACKGROUND： Inflammatory factor is closely associated with the function of endothelial cells. Studies on effects of protective and injured factors on inflammatory factors of in vitro cultured human vascular endothelial cells are few. OBJECTIVE： In this study, we observed the influence of curcumine and tumor necrosis factor （TNF- α ） on nuclear factor- κB （NF- κB） and monocyte chemotactic protein-1 （MCP-1） in human umbilical vein endothelial cells. DESIGN, TIME AND SETTING： A cell observational experiment was performed at the Laboratory of Tissue Engineering of Fourth Military Medical University of Chinese PLA from December 2006 to August 2007. MATERIALS： Healthy human umbilical vein was provided by Tangdu Hospital of Fourth Military Medical University of Chinese PLA. Endothelial cells were inoculated in M199 complete medium. METHODS： The experiment included 3 groups. Endothelial cells were cultured in serum-free M199 medium in the normal control group. 10 μ g/L TNF-α was added in the normal culture medium for 1.5 hours in the TNF-α group. Endothelial cells were inoculated in medium supplemented with 10 μ g/L TNF-α for 30 minutes, and then incubated with different dosages of curcumine （6.25, 12.5, 25, 50, 100 mg/L） for 1 hour in the curcumine group. MAIN OUTCOME MEASURES： Different dosages of curcumine effects on the proliferation of human umbilical vein endothelial cells by methyl thiazolyl tetrazolium （MTT）. NF-κB expression was examined by immunocytochemistry. MCP-1 mRNA expression was detected were measured by reverse transcription-polymerase chain reaction （RT-PCR）. RESULTS： Curcumine could improve the activity of human umbilical vein endothelial cells after injury by TNF-α （P 〈 0.01）, with dose-dependent effects to some extent. TNF-α could stimulate the expression of NF- κBp65. Curcumine could decrease the expression of NF- κBp65. Compared with the normal control group, levels of MCP- 1 mRNA significantly increased after TNF-α intervention f

关 键 词：核转录因子ΚB 姜黄素 人脐静脉内皮细胞 肿瘤坏死因子Α 单核细胞趋化蛋白1 

分 类 号：R392[医药卫生—免疫学]