结核分枝杆菌氧氟沙星耐药和敏感临床分离株的定量蛋白质组学分析  

Quantitative proteomic analysis of ofloxacin resistant and sensitive clinical isolates of Mycobacterium tuberculosis

作　　者：黄香玉[1] 朱传智[1] 逄宇[2] 邵进士[1] 张春青[1] 何秀云[1] 赵雁林[2] 庄玉辉[1] 

机构地区：[1]解放军309医院器官移植与免疫调节北京市重点实验室,北京1100091 [2]中国疾病预防控制中心结核病预防控制中心,国家结核病参比实验室,北京102206

出　　处：《解放军医学杂志》2014年第9期703-708,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家重要传染病科技重大专项(2008ZX10003-009)~~

摘　　要：目的探讨与结核分枝杆菌(MTB)氧氟沙星(OFX)耐药相关的蛋白。方法苏通培养基培养MTB H37Rv标准株,临床分离MTB OFX耐药株(OFXR)和敏感株(OFXS),菌体经60钴照射灭活提取菌体蛋白。菌体蛋白采用iTRAQ技术和生物信息学鉴定并相对定量。结果 MTB OFXR分离株与MTB OFXS分离株和H37Rv标准株比较分别有175个和134个差异表达蛋白,OFXR/OFXS与H37Rv的共同差异表达蛋白有104个。差异表达蛋白理论相对分子量和等电点分布广泛,主要参与脂质代谢、中间代谢和呼吸作用。9个共同差异表达蛋白在OFXR分离株中呈现显著差异(差异比值>1.2或<0.55),其中Rv0106、Rv0895、Rv2185c、Rv3248c和Rv3841表达上调,而Rv2524c、Rv2986c、Rv3118和Rv3597c表达下调。结论利用iTRAQ技术发现了MTB OFXR分离株与MTB OFXS分离株和H37Rv标准株的共同差异表达蛋白,为进一步探讨MTB对OFX耐药的机制提供了基础。Objective To identify the proteins related to ofloxacin （OFX） resistance of Mycobacterium tuberculosis （MTB）. Methods Standard MTB H37Rv strain, clinical isolates of OFX resistant strain （OFXR） and sensitive strain （OFXs） were obtained from the Chinese Center for Disease Control and Prevention, and they were cultured in Sauton＇s medium, and then inactivated by ^60 Co. Whole cellular proteins were extracted from OFXR, OFXs and H37Rv strain of MTB, respectively. The peptides were labeled, separated and identified by isobaric tags of relative and absolute quantitation （iTRAQ） combined with Nano LC- MS/MS technology. Results One hundred and seventy-five and 134 differential expression proteins were identified in MTB OFXR compared with MTB OFXs and H37Rv, respectively. One hundred and four common differential expression proteins were identified in MTB OFXR compared with both MTB OFXs and H37Rv. The isoelectric point and theoretic relative molecular mass of differential expression proteins were widely distributed. The majority of the common differential expression proteins were involved in intermediary metabolism, respiration, and lipid metabolism. Twelve common differential expression proteins showed significant differences （the ratio〉l.2 or 〈0.55） in MTB OFXg, including Rv0106, Rv0895, Rv2185c, Rv3248c and Rv3841 up-regulation and Rv2524c, Rv2986c, Rv3118 and Rv3597c down-regulation. Conclusion iTRAQ has been used to identify the common differential expression proteins in MTB OFXR compared with both MTB OFXs and H37Rv, which provides a basis for further study of the mechanism of OFX-resistance.

关 键 词：结核 抗多种药物性 氧氟沙星 蛋白质组学 

分 类 号：R378.911[医药卫生—病原生物学]