锌指蛋白A20在灵仙新苷减轻血脑屏障炎性损伤中的作用及其机制研究

项目名称： 锌指蛋白A20在灵仙新苷减轻血脑屏障炎性损伤中的作用及其机制研究

项目编号： No.81202974

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学八处

项目作者： 方伟蓉

作者单位： 中国药科大学

项目金额： 23万元

中文摘要： 基于前期工作及文献提出上调锌指蛋白A20表达、负反馈抑制NF-κB，是灵仙新苷减轻血脑屏障炎性损伤的重要机制。灵仙新苷是威灵仙的单体成分，前期研究表明有显著的抗类风湿性关节炎作用；我们进一步研究发现，灵仙新苷能减轻关节炎大鼠血脑屏障损伤，且能上调TNF-α刺激的脑微血管内皮细胞A20表达。本项目在前期工作基础上，建立锌缺乏合并脑缺血模型，确证补锌作用是灵仙新苷保护血脑屏障完整性的物质基础；并通过体外试验验证锌缺乏致脑微血管内皮细胞A20表达抑制、介导炎症反应的作用，进而阐明灵仙新苷上调A20表达的机制；在此基础上，运用基因转染、Western等分子生物学技术，阐明灵仙新苷通过上调A20表达、负反馈抑制TNF-α→TNFR1→NF-κB信号通路、抑制血脑屏障炎性损伤的分子机制。研究成果将为灵仙新苷的临床应用提供依据，并对基于锌指蛋白A20的抗炎靶点研究具有借鉴作用。

中文关键词： 灵仙新苷；血脑屏障；炎症；脑缺血/再灌注；锌指蛋白A20

英文摘要： Based on previous research and references, we proposed that Clematichinenoside could protect blood brain barrier (BBB) inflammatory injury through upregulation of zinc finger protein A20 and inhibition of NF-κB. Clematichinenoside was one monomer of clematis chinensis osbeck that could obviously inhibit rheumatoid arthritis (RA). Also in our early stage work, BBB integrity protection in RA rats and zinc finger protein A20 upregulation in brain microvessel endothelial cells were observed after administration or inbubation with Clematichinenoside. In this project, zinc deficiency combine with cerebral ischemia model of rats will be established to study zinc supplement and BBB injury inhibition effect of Clematichinenoside. In vitro experiments, brain microvessel endothelial cells were served as in vitro model of BBB. Culture medium with high Zn2+ or Zn2+ deprivation to verify Zn2+ deficiency could mediate NF-κB oevrexpression and inflammation which induced by A20 supression. The results will reveal that mechanism of Clematichinenoside on upregulation A20 was depending on zinc supplement. Furthermore, we will apply gene transfection and western technology to illustrate the molecular mechanism of Clematichinenoside on inducing A20 expression and negatively inhibiting NF-κB through TNF-α→TNFR1→NF-κB signal pathway. T

英文关键词： Clematichinenoside；Blood brain barrier；Inflammation；Ischemic stroke；Zinc finger protein A20