寡核苷酸芯片研究结核分枝杆菌临床株和无毒株诱导的巨噬细胞U937凋亡相关基因的差异表达  

Differential Expression of Apoptosis-related Gene Induced by Clinical and Laboratory Mycobacterium tuberculosis Strain in Macrophages U937 Revealed by Oligonucleotide Microarray

作　　者：胡昌华[1] 谢建平[1,2] 李瑶[2] 乐军[3] 徐永忠[2] 王洪海[2] 

机构地区：[1]西南师范大学现代生物医药研究所生命科学学院,重庆400715 [2]复旦大学生命科学学院遗传学研究所病原生物学组,遗传工程国家重点实验室,上海200433 [3]上海市肺科医院,上海200433

出　　处：《Acta Genetica Sinica》2004年第3期231-235,共5页

基　　金：国家重大基础研究规划项目 (编号 :G 1 990 541 0 4 );国家自然科学基金(编号 :30 1 0 0 0 7)资助~~

摘　　要：结核病仍然是人类健康的主要威胁 ,结核分枝杆菌诱导的巨噬细胞凋亡是宿主防御反应之一 ,研究凋亡相关基因的差异表达有助于认识结核分枝杆菌致病机理和发现新的药物靶标。利用包括 192 0 0个基因或基因片段的DNA芯片研究巨噬细胞株U93 7对临床和实验室菌株感染的差异表达 ,Northernblotting和RT PCR验证了芯片研究结果。MtbH3 7Rv感染下调bcl 2 ,vitaminD受体、干扰素调控因子 3、细胞色素氧化酶C表达 ,幅度分别为 2 ,3 ,3 ,2 5 倍 ,临床菌株感染上调SOD2、SOD3、丝氨酸蛋白酶、toll like受体 2、signaltransducerandactivator (STAT1)、hy poxia induciblefactor 2 2等表达 ,幅度分别为 2 9 ,2 5 ,2 5 ,2 2 ,2 4 ,5 9 倍。结果提示 ,临床菌株感染更多促进凋亡 ,限制宿主的杀灭机理。Tuberculosis(TB) remains one of the major problems in global health.Macrophage (MPhi) apoptosis,induced by Mycobacterium tuberculosis (Mtb),is a cornerstone of effective innate microbial defense mechanism.Elucidation of the complex apoptosis-related gene expression may facilitate understanding the mechanism and regulation of macrophage apoptosis in response to Mtb,and contribute to developing novel measures to counter TB.DNA microarray containing 19 200 gene or gene fragments was used to compare the macrophage cell line U937 gene expression response to the clinical and laboratory Mtb infection.Northern blotting and RT-PCR were used to confirm the microarray results.Mtb H37Rv infection were found to downregulate the bcl-2,vitamin D receptor,interferon regulatory factor 3,cytochrome c oxidase,gene expression by 2-,3-,3-,2.5-fold,respectively,while the clinical strain infection leads to upregulate the SOD2,SOD3,serine protease,toll-like receptor 2,signal transducer and activator (STAT1),hypoxia-inducible factor 22,2.9-,2.5-,2.5-,2.2-,2.4-,5.9-fold respectively.The findings suggest that the clinical strain infection tends to override the macrophage apoptosis by which the host attempt to limit the growth of the invader.The research on the complex factors network involved in the interaction will benefit the vaccine and novel drug target development.

关 键 词：凋亡 MYCOBACTERIUM TUBERCULOSIS 巨噬细胞 

分 类 号：R346[医药卫生—基础医学]