结核分枝杆菌诱导的THP-1细胞转化及其相关信号传导通路的研究  

Determination of cellular transformation of THP-1 cells mediated by Mycobacterium tuberculosis spp. infection and the involved signaling pathways

作　　者：秦妍妍[1] 黄晓燕 李淑萍[3] 李立伟[1] 毛亚飞[1] 严杰[3] 

机构地区：[1]浙江大学医学院病原生物学系,杭州310058 [2]浙江省宁波市李惠利医院 [3]浙江大学医学院附属第一医院传染病诊治国家重点实验室

出　　处：《中华微生物学和免疫学杂志》2008年第5期385-389,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目（30400018）

摘　　要：目的了解结核分枝杆菌（Mycobacterium tuberculosis）体外诱导人单核细胞株THP-1向类上皮细胞（EC）转化的效应及其相关信号传导通路和调控作用。方法建立人结核分枝杆菌H37Rv株、牛分枝杆菌bovis株、草分枝杆菌phlei株THP-1细胞感染模型。采用间接免疫荧光法检测感染前后THP-1细胞表面单核细胞或巨噬细胞分化抗原CD115和EC分化抗原CD82的表达情况。采用Sandwich ELISA Kits检测感染前后THP-1细胞p38MAPK（p38丝裂原活化蛋白激酶）、Akt1（丝／苏蛋白激酶）和STAT3（信号转导因子和转录活化因子）磷酸化水平。采用特异性阻断剂，了解各信号通路阻断前后CD115和CD82表达水平变化。结果3株分枝杆菌感染的THP-1细胞均出现CD115表达减弱和CD82明显表达的现象。H37Rv株或bovis株感染的THP-1细胞可出现一过性p38MAPK磷酸化水平上调，但P13K（磷脂酰肌醇3-激酶）／Akt和STAT3磷酸化水平均无明显变化。p38MAPK、P13K／Akt或JAK／STAT通路被阻断后，上述3株分枝杆菌感染的THP-1细胞仍表达CD115。JAK／STAT通路被阻断时，各分枝杆菌感染的THP-1细胞仍表达CD82。但p38MAPK、P13K／Akt通路被阻断时，H37Rv株和bovis株感染的THP-1细胞CD82表达消失。结论结核分枝杆菌H37Rv株和bovis株感染后可诱导THP-1细胞向EC转化，p38MAPK、P13K／Akt参与和调控了该转化过程，其中以p38MAPK通路较为重要。Objective To determine the effect of Mycobacterium tuberculosis slap. inducing transformarion of THP-1 cells to epithelioid cells （EC） and the involved signaling pathways and their regulations. Methods THP-1 cells infection models respectively infected with M. tuberculosis strains H37Rv, bovis and phlei were established. Indirect immunofluorescent staining assays were used to detect the expressions of monocyte/macrophage differentiation antigen CD115 and EC differentiation antigen CD82 of the THP-1 cells before or after infection. By Sandwich ELISA Kits, the phosphorylation levels of p38MAPK, Aktl and STAT3 of the THP- 1 cells before or after infection were measured. The alterations of CDll5 and CD82 expression levels were examined when the associated signaling pathways were blocked with specific blocking agents. Results CD115 expression was weakened and CD82 expression was strongly increased in all the THP-1 cells infected with the three strains. A temporal up-regulation of the p38MAPK phoshporylation level but no obvious alteration of Akt and STAT3 phosphorylarion levels after THP-1 cells infected by strain H37Rv or boris. The THP-1 cells infected with anyone of the three strains continuously expressed CD115 after MAPK, PI3K/Akt or JAK/STAT of the cells was blocked. Although JAK/STAT was blocked, the THP-1 cells respectively infected with the three different strains still expressed CD82. However, CD82 expressed in THP-1 cells infected by the strain H37Rv or bovis was disappeared when p38MAPK and PI3K/Akt pathways of the cells were blocked. Conclusion Strain H37Rv and bovis can induce the infected THP-1 cells transforming to EC and p38MAPK and PI3K/Akt signaling pathways participate and regulate the transformation procedure. Of the two signaling pathways p38MAPK seems to be more important.

关 键 词：结核分枝杆菌 细胞转化 分化抗原表达 信号通路/调控 

分 类 号：R378[医药卫生—病原生物学] R52[医药卫生—基础医学]