| 利用生物素-链亲和素系统筛选呼吸机所致肺损伤HMGB1启动子结合蛋白 |
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| 引用本文： | 丁宁,肖慧,许立新,佘守章. 利用生物素-链亲和素系统筛选呼吸机所致肺损伤HMGB1启动子结合蛋白[J]. 医学争鸣, 2009, 0(21): 2345-2347 |
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| 作者姓名： | 丁宁 肖慧 许立新 佘守章 |
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| 作者单位： | 广州市第一人民医院麻醉科和麻醉学实验室;广州市第一人民医院门诊部; |
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| 基金项目： | 广东省医学科研基金项目(A2009497); 广州市医药卫生科技一般引导项目(2008-YB-012,2009-YB-021); 广州市医药卫生科技重点项目(2008-ZDi-14) |
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| 摘 要： | 目的：筛选呼吸机所致肺损伤（VILI）高迁移率族蛋白B1（HMGB1）的启动子结合蛋白.方法：制备VILI大鼠模型,与正常对照组大鼠同期处死,取肺组织提取细胞核蛋白.用PCR法扩增末端带生物素标记的HMGB1启动子探针,将细胞核蛋白与制备的HMGB1启动子探针进行孵育,再以标记有链亲和素的磁珠分离HMGB1启动子-蛋白反应复合物,分别用0.25mol/L和1mol/LNaCl洗脱探针上结合的蛋白,SDS-PAGE电泳分离样品蛋白,并对凝胶进行银染显色,比较VILI组与正常对照组的差异条带.结果：两组细胞核蛋白中共观察到24条差异条带.分析差异条带显示,与正常对照组比较,在低亲和力作用水平,机械通气后有6个蛋白与HMGB1启动子的结合力增强,有2个蛋白的结合力减弱;在高亲和力作用水平,有10个蛋白与HMGB1启动子的结合力增强,有6个蛋白的结合力减弱.结论：筛选到一些VILI特异性HMGB1启动子结合蛋白,对于研究HMGB1的转录调控机制具有重要意义.
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| 关 键 词： | 高迁移率族蛋白B1 机械通气 急性肺损伤 启动子 生物素-链亲和素 |
Screening of binding proteins that interact with HMGB1 promoter in ventilator-induced lung injury by biotin-streptavidin system |
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| DING Ning,XIAO Hui,XU Li-Xin,SHE Shou-Zhang. Screening of binding proteins that interact with HMGB1 promoter in ventilator-induced lung injury by biotin-streptavidin system[J]. Negative, 2009, 0(21): 2345-2347 |
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| Authors: | DING Ning XIAO Hui XU Li-Xin SHE Shou-Zhang |
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| Affiliation: | DING Ning1,XIAO Hui2,XU Li-Xin1,SHE Shou-Zhang11Department of Anesthesiology and Anesthesiology Lab,2Department of Out-patient,Guangzhou First Municipal People's Hospital,Guangzhou 510180,China |
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| Abstract: | AIM:To screen the binding proteins that interact with HMGB1 promoter in ventilator-induced lung injury(VILI). METHODS:Eight Sprague Dawley(SD) rats were divided into two groups,the mechanically ventilated group(n=4) and the normal control group(n=4). The mechanically ventilated rats were subjected to mechanical ventilation(VT=40 mL/kg) for 4 hours to reproduce a ventilator-induced lung injury(VILI) model. The biotin-labeled DNA probe for HMGB1 promoter was amplified by PCR. The cell nuclear extracts from lu... |
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| Keywords: | high mobility group box 1 protein mechanical ventilation acute lung injury promoter biotin-streptavidin |
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