课题背景：课题受辽宁省教委资助（05L246）。项目名称：桃核乘气改良方对家兔血管球囊损伤术后血浆脑利钠肽和信号转导及转录激活因子3影响的实验研究。

应用要点：实验结果提示，桃核乘气改良方能有效地降低冠心病介入治疗技术术后兔血中内皮素的含量，升高一氧化氮含量，对内皮素、一氧化氮分泌具有调整治疗作用，促进两者之间的平衡协调；抑制血管内皮增生，促进损伤内膜修复。

同行评价：实验通过建立动脉粥样硬化模型，对比分析各组内皮素和一氧化氮含量差异，为中药桃核乘气改良方促进血管内皮细胞正常生长增生提供实验依据。

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1 mRNA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10 μg/L 的肿瘤坏死因子α诱导损伤1.5 h；姜黄素组：先在培养液中加入10 μg/L 的肿瘤坏死因子α30 min 诱导损伤，再分别加入终浓度为6.25，12.5，25，50，100 mg/L 的姜黄素孵育1 h。
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1 mRNA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30 min 后，单核细胞趋化蛋白1 mRNA相对表达量明显增高（P < 0.01）；加入不同浓度的姜黄素作用1 h 后，单核细胞趋化蛋白1 mRNA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1 mRNA的表达，从而阻滞了单核细胞的聚集。
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

刘镘利，王宗仁，张筠，雷战军. 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1 mRNA的变化[J].中国组织工程研究与临床康复，2008，12(20):3841-3844 [www.zglckf.com/zglckf/ejournal/upfiles/08-20/20k-3841(ps).pdf]

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

Abstract

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 μg/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 μg/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 for 30 minutes (P < 0.01), but reduced after different dosages of curcumine intervention for 1 hour (P < 0.05，P < 0.01).
CONCLUSION: Curcumine displays a protective action on injured endothelial cells. Curcumine prevent the aggregation of mononuclear cells by inhibiting NF-κb activity and depressing MCP-1 mRNA expression.

Liu ML, Wang ZR, Zhang Y, Lei ZJ. Changes in nuclear factor kappa B and monocyte chemotactic protein 1 mRNA during the process of human umbilical vein endothelial cells injury.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(20):3841-3844(China) [www.zglckf.com/zglckf/ejournal/upfiles/08-20/20k-3841(ps).pdf]