HSP60通过Toll样受体对ApoE^(-/-)小鼠树突状细胞功能影响研究  

Effect of heat shock protein 60 on the activation of dendritic cells by Toll-like receptors in ApoE^(-/-) mice

作　　者：吴伟[1] 蒋振忠[1] 周游[1] 

机构地区：[1]南京医科大学第二附属医院心内科,南京210011

出　　处：《中国免疫学杂志》2010年第6期498-502,共5页Chinese Journal of Immunology

基　　金：2007年度南京医科大学科技发展基金面上项目资助

摘　　要：目的:探讨HSP60通过Toll样受体(TLR)介导丝裂原活化蛋白激酶(MAPK)信号转导在体外对ApoE-/-小鼠树突状细胞(mDC)的功能影响。方法:将ApoE-/-小鼠构建动脉粥样硬化模型后,从骨髓提取DC,体外培养成熟后分三组:HSP组、LPS组及对照组,分别与HSP60、LPS及生理盐水孵育后,应用流式细胞仪检测细胞表面TLR及CD80水平,RT-PCR检测TLR的mRNA水平,免疫印迹法检测MAPK家族中p38及ERK水平;上清液用ELISA法检测IL-6及TNF-α水平。结果:与对照组比较,HSP组中DC表达TLR及MAPK家族水平、成熟标记物CD80水平、分泌IL-6及TNF-α水平均明显升高(P<0.01);但与LPS组比较则无显著差异(P>0.05)。结论:HSP60通过TLR介导MAPK信号转导参与动脉粥样硬化模型mDC的活化过程。Objective：To investigate the effect of heat shock protein 60（HSP60） on the functions of dendritic cells of ApoE-/-mice （mDCs） in vitro by signal transduction of mitogen-activated protein kinase （MAPK） family mediated by Toll-like receptors （TLRs）.Methods：Myeloid mDCs were extracted in vitro from ApoE-/-mouse marrow,which were incubated as atherosclerotic animal model,and divided into three groups：HSP group,LPS group and control group.The three groups of mDCs were incubated to mature with mHSP60,LPS and normal saline,respectively and then examined the expression of TLR protein and mRNA by flow cytometry and real-time RT-PCR.In addition,the expression level of CD80 was examined by flow cytometry.The expression of MAPK family proteins was determined by Western blot analysis.The levels of cytokines secreted by mDCs,such as IL-6 and TNF-α,were analyzed by ELISA.Results：The expression of TLRs and MAPK families （p38 and ERK） was increased in the samples of HSP group,when compared with control group （P0.01）.Likewise,the levels of the mature marker （CD80） and cytokines secreted （IL-6 and TNF-α） were elevated in HSP group （P0.01）.However,there was no significant deviation compared to that of LPS group （P0.05）.Conclusion：HSP60 is involved in activation of mDCs of atherosclerotic animal model by means of the signal transduction of MAPK family mediated by TLRs.

关 键 词：TOLL样受体 丝裂原活化蛋白激酶 热休克蛋白60 树突状细胞 信号传导 

分 类 号：R392[医药卫生—免疫学]