内毒素刺激大鼠腹腔巨噬细胞高迁移率族蛋白B1释放的研究  

Lipopolysaccharide induces high mobility group box-1 protein release in peritoneal macrophages in rats

作　　者：刘辉[1] 姚咏明[1] 董月青[1] 盛志勇[1] 

机构地区：[1]解放军总医院第三O四临床部全军烧伤研究所基础部,北京100037

出　　处：《中华实验外科杂志》2005年第1期37-38,共2页Chinese Journal of Experimental Surgery

基　　金：国家重点基础发展规划项目 (G1 9990 542 0 3) ;国家杰出青年科学基金资助项目 (30 1 2 50 2 0 )

摘　　要：目的　观察高迁移率族蛋白B1(HMGB1)在大鼠巨噬细胞中的分布和释放。方法　取正常Wistar大鼠腹腔巨噬细胞 ,培养 3d后以内毒素 (LPS)刺激 ,应用免疫组织化学的方法检测LPS刺激前后HMGB1的表达释放情况 ;分正常、4、8、12、2 4h时间点留取细胞培养上清 ,观察LPS刺激与HMGB1释放的时效关系。结果　大鼠腹腔巨噬细胞的胞核及胞浆内存在HMGB1蛋白表达 ,LPS刺激 10h使之明显增强 ,并分泌至胞外 ;LPS可诱导大鼠腹腔巨噬细胞HMGB1释放 ,于 8～ 12h达峰值 (正常对照和 12h的积分吸光度值分别为 19.3 99± 1.488、5 9.5 61± 2 .991,P <0 .0 1) ,2 4h后减弱 (积分吸光度值为 2 6.10 4± 2 .671)。结论　HMGB1分布于大鼠腹腔巨噬细胞核及胞浆中 ,LPS可促进HMGB1缓慢释放于细胞外。Objective To observe high mobility group box 1 protein (HMGB1) expression and release in rat peritoneal macrophages stimulated with lipopolysaccharide (LPS).Methods Peritoneal macrophages obtained from male Wistar rats were incubated for 3 days before they were stimulated with LPS.After stimulation,HMGB1 protein expression as well as release before and after stimulation was detected by immunohistochemistry.Supernatants of controls,4 h,8 h,12 h and 24 h were collected to determine the time dependent response between LPS stimulation and HMGB1 release.Results HMGB1 was expressed in nucleus and cytoplasm of rat peritoneal macrophages.LPS stimulation could markedly enhance the HMGB1 expression and release.LPS induced HMGB1 release in rat peritoneal macrophages,peaking at 8 12 h (optical density values of normal controls vs.12 h: 19.399± 1.488 vs. 59.561± 2.991, P < 0.01) and decreasing 24 h later (optical density value: 26.104± 2.671). Conclusion HMGB1 exists in nucleus and cytoplasm of rat peritoneal macrophages,which can release extracellularly by LPS stimulation.

关 键 词：腹腔巨噬细胞 LPS 大鼠 刺激 高迁移率族蛋白B1 正常 HMGB1 释放 分泌 细胞培养 

分 类 号：R363[医药卫生—病理学]