穿透支原体脂质相关膜蛋白激活核因子κB诱导小鼠巨噬细胞表达诱导性一氧化氮合酶  

Expression of inducible nitric oxide synthase in murine macrophages stimulated with Mycoplasma penetrans lipid-associated membrane proteins by activating nuclear factor κB

作　　者：曾焱华[1] 吴移谋[1] 余敏君[1] 朱翠明[1] 曾铁兵[1] 刘安元[1] 

机构地区：[1]南华大学医学院微生物学与免疫学教研室,衡阳421001

出　　处：《中华微生物学和免疫学杂志》2004年第10期807-811,共5页Chinese Journal of Microbiology and Immunology

基　　金：湖南省自然科学基金 ( 0 2JJY2 0 2 5 )

摘　　要：目的　了解穿透支原体潜在的致病性和穿透支原体脂质相关膜蛋白诱导小鼠巨噬细胞表达诱导性一氧化氮合酶 (iNOS)的分子机制。方法　用从穿透支原体提取的脂质相关膜蛋白刺激小鼠巨噬细胞 ,以RT PCR和Westernblot等方法分析诱导性一氧化氮合酶的表达和NO的产生。用间接免疫荧光和Westernblot检测经穿透支原体脂质相关膜蛋白处理的小鼠巨噬细胞中NF κB的激活和NF κB抑制剂PDTC(吡咯啉烷二甲基硫脲 )对iNOS的表达和NO产生的影响。结果　穿透支原体脂质相关膜蛋白以时间和剂量依赖方式刺激小鼠巨噬细胞产生NO ,且能激活NF κB诱导巨噬细胞表达i NOS的mRNA和蛋白 ,其mRNA和蛋白的表达水平能被PDTC所抑制。结论　由于穿透支原体脂质相关膜蛋白能通过激活NF κB诱导巨噬细胞表达诱导性一氧化氮合酶 ,因而可能是一个重要的致病因素。Objective To investigate the potential pathogenicity of Mycoplasma penetrans(M. penetrans) and the molecular mechanisms that is responsible for the induction of inducible nitric oxide synthase(iNOS) gene expression in mouse macrophages stimulated by lipid-associated membrane proteins(LAMPs) prepared from M. penetrans. Methods Mouse macrophages were stimulated by M. penetrans LAMPs to analyze the production of nitric oxide(NO), the expression of iNOS was detected by RT-PCR and Western blot. The activity of nuclear factor κB(NF-κB) and the effects of pyrrolidine dithiocarbamate(PDTC), an inhibitor of NF-κB, on the production of nitric oxide and the expression of iNOS was also examined in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blot. Results M. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. mRNA and protein of iNOS were also induced in response to LAMPs and inhibited by PDTC. M. penetrans LAMPs could trigger NF-κB activation, a possible mechanism for the induced iNOS expression. Conclusion M. penetrans may be an important etiological factor due to the ability of LAMPs to stimulate the expression of iNOS, which is probably mediated by the activation of NF-κB.

关 键 词：巨噬细胞 支原体 脂质 表达 小鼠 一氧化氮合酶 膜蛋白 RNA 体脂 NF-κB 

分 类 号：R375[医药卫生—病原生物学]