牛分枝杆菌感染巨噬细胞(THP-1)全基因表达谱变化的研究

The profile of gene expression from macrophage THP-1 infected by Mycobacterium bovis

目的分析可用于牛结核病免疫学诊断的候选靶标,同时初步探讨由牛分枝杆菌(Myco-bacterium bovis)引起人结核病的免疫学发病机制。方法牛分枝杆菌(AF 2212/97)和结核分枝杆菌(Mycobacterium tuberculosis(H37Rv))感染人的模式巨噬细胞(THP-1),采用含有22000个基因全长的基因芯片分析感染72h后的THP-1细胞全基因组的在转录水平的表达谱变化。在确定所分析的芯片的特异性和稳定性均达到要求的基础上,通过GO显著性统计分析,研究表达变化的基因所对应蛋白的分子功能,生物过程以及细胞组分;进一步采用Pathway显著性统计分析,研究表达差异蛋白之间的相互作用。结果感染H37Rv后的THP-1细胞,存在290个差异基因表达,其中170个上调表达,120个下调表达,与以往的结果类似。牛分枝杆菌感染的THP-1细胞中有1899(8.6%)个基因差异表达,其中1075个基因被上调表达(56.6%),其他43.4%被下调。1899个基因中1674个在GenBank有记录,263个是新基因,,基质金属蛋白酶12(H200000442)被上调达60.4921倍;下调幅度最大的蛋白丝氨酸/半胱氨酸抑制酶(H200006177)被下调6.29倍。采用GO分析发现,这些基因对应的多为炎症因子蛋白、凋亡蛋白、细胞外基质蛋白、T细胞受体信号通路相关蛋白等。其中与炎症相关的的蛋白有47个,如TNF、IL8、CXCL3、CCL8、CCL7、IFNK、IL-26、CCL28等。随后的Path-way分析差异表达基因对应的2014个蛋白,展示了一系列的引起结核病发病的重要信号传导通路。结论基因表达谱研究可为牛结核病发病机制及其诊断研究提供大量有益的生物学信息。

Objective To select the candidate target for immunological diagnosis of bovine tu- berculosis, and to study the immunological pathogenetic mechanisms of bovine tuberculosis caused by Mycobacterium bovis. Methods The gene expression profiles from macrophage THP-1 infected by Mycobacterium boris （AF2212/97） and Mycobacterium tuberculosis （H37Rv） 72 hours later were analyzed with gene chips containing 22 000 full-length genes. The protein expression level and their interaction of corresponding gene were compared and analyzed by GO and Pathway significant statistical analysis. Results There were expression difference of 290 and 1899 genes in maerophage THP-1 cells infected by Mycobacterium tuberculosis and Mycobacterium bovis, respectively. These dif- ferential genes mainly included inflammatory factors, apoptosis proteins, extracellular matrix protein, the proteins correlating with T-cell receptor signaling pathway, and other related proteins by GO and Pathway analysis. Conclusions The result of gene expression profile could provide the useful biology information for the diagnostic and pathogenetic study of bovine tuberculosis.