细菌脂多糖、脂蛋白和DNA对小鼠肺泡巨噬细胞体外激活的协同作用  

The synergistic effects of lipopolysaccharide, bacterial lipoprotein and bacterial DNA on mouse alveolar macrophage activation

作　　者：黄宏[1] 蒋建新[1] 朱佩芳[1] 王正国[1] 张道杰[1] 杨程[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所第四研究室全军交通医学研究所创伤,烧伤与复合伤国家重点实验室,重庆400042

出　　处：《中华医学杂志》2005年第21期1468-1472,共5页National Medical Journal of China

基　　金：国家重点基础研究专项经费资助项目(G1999054203);国家杰出青年科学基金资助项目(30325040)

摘　　要：目的通过体外实验,从受体水平观察细菌脂多糖(LPS)、细菌脂蛋白(BLP)和细菌DNA对肺泡巨噬细胞的协同刺激效应及其可能机制。方法分离培养小鼠肺泡巨噬细胞,随机将细胞分为7组,分别为LPS组、CpGODN组、BLP组、LPS+CpGODN组、LPS+BLP组、LPS+CpGODN+BLP组和培养基对照组。刺激6h后,收集上清和细胞,上清用于TNFα检测,细胞用于提取总RNA,通过RTPCR检测主要模式识别受体(PRRs)CD14、SR、TLR2、TLR4、TLR9的表达。结果LPS、BLP和CpGODN不仅体外能协同增加肺泡巨噬细胞释放TNFα等细胞因子,而且具有协同增强效应细胞表面模式识别受体的表达,以三者同时存在时,协同作用最强。结论细菌内毒素、细菌脂蛋白和细菌DNA在体外不仅能上调相互的模式识别受体,而且具有协同增强其他细菌结构成分对相应受体的刺激作用。Objective To investigate the synergistic effects of lipopolysaccharide (LPS), bacterial lipoprotein (BLP), and bacterial DNA on the expression of pattern recognition receptors (PRRs) on the cell surface of mouse alveolar macrophages and cellular activation at the level of receptor and its possible mechanism Methods Mouse alveolar macrophages were isolated, cultivated and randomly divided into 7 groups: control group, LPS group, CpG oligonucleoetide (CpG-ODN) group, BLP group, LPS+BLP group, LPS+CpG-ODN group, and LPS+BLP+CpG-ODN group. Six hours later the supernatants were collected to detect the level of tumor growth factor α (TNFα) by ELISA. RT-PCR was used to detect the expression of the main PRRs: CD14, SR, TLR2, TLR4, and TLR9. Results The TNFαlevels in the supernatant were 234 pg/ml±30 pg/ml in the LPS group, 274 pg/ml±30 pg/ml in the BLP group, and 308 pg/ml±28 pg/ml in the CpG-ODN group, all significantly higher than that in the control group (92 pg/ml±27 pg/ml, P<0.01 or P<0.01). the TNFα levels in the supernatant were 483 pg/ml±31 pg/ml in the LPS+BLP group, and 511 pg/ml±46 pg/ml in the LPS+CpG-ODN group, both significantly higher than those of the groups of the 3 factor alone (all P<0.05). And the TNFαlevels in the supernatant was 665 pg/ml±24 pg/ml in the LPS+BLP+CpG-ODN group, significantly higher than those of the LPS+ ODN group and LPS+BLP group(both P<0.05). LPS, BL, and CpG-ODN alone, combinations of any 2 of them, and the combination of the three all up-regulated the expression of CD14 mRNA more and more strongly in sequence. LPS, BLP, and CpG-ODN alone all up-regulated the expression of SR mRNA (all P<0.01), however, the combinations of any 2 factors or of the 3 factors failed to further up-regulate the expression of SR. LPS and BLP up-regulated the expression of TLR2 mRNA (both P<0.05), LPS combined with BLP showed a stronger up-regulation of TLR2 mRNA (P<0.05) than those by LPS and BLP alone. CPG-ODN alone failed to up-regulate the expression of TLR2 mRNA (P>0.05)but significantl

关 键 词：肺泡巨噬细胞 协同作用 脂蛋白 细菌脂多糖 体外激活 小鼠 CPG-ODN 模式识别受体 脂多糖(LPS) RT-PCR检测 细菌DNA TNF-α 协同增强效应 细菌内毒素 体外实验 受体水平 可能机制 刺激效应 分离培养 总RNA TLR4 TLR9 细胞因子 

分 类 号：R563[医药卫生—呼吸系统]