Sigma因子基因表达与结核分枝杆菌异烟肼耐药关系探讨  

Exploration of the relationship between Sigma factor gene expression and isoniazid resistance in Mycobacterium tuberculosis

作　　者：江丽娜[1] 高丽 王志锐[1] 王秀月 王春花[1] JIANG Lina;GAO Li;WANG Zhirui;WANG Xiuyue;WANG Chunhua(Tianjin Key Laboratory of Pathogenic Microbiology of Infections Disease,Tianjin Center for Tuberculosis Control,Tianjin 300011,China)

机构地区：[1]天津市病原微生物重点实验室,天津市结核病控制中心,天津300011

出　　处：《中国热带医学》2024年第3期299-303,共5页China Tropical Medicine

基　　金：天津市卫生健康科技项目(No.TJWJ2022MS047);天津卫生健康行业高层次人才选拔培养工程项目(No.TJSJMYXYC-D2-017)。

摘　　要：目的研究调控基因Sigma因子(sigA-sigM)在katG突变导致的异烟肼耐药结核分枝杆菌(Mycobacterium tuberculosis,MTB)中的表达调控是否与异烟肼耐药相关,为研究异烟肼耐药的分子机制提供参考依据。方法选择2020—2022年天津市结核病控制中心耐药检测期间收集的初治患者菌株共95株,其中katG基因未突变的全敏菌株30例,katG突变导致的异烟肼耐药菌株共65例,包括11例单耐异烟肼菌株,24株异烟肼多耐药菌株(同时耐异烟肼和链霉素),30例异烟肼耐多药菌株(同时耐异烟肼、链霉素和利福平)。在异烟肼含药培养基上收集菌株后,提取菌株RNA,应用反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)方法检测Sigma因子的相对表达量,采用Mann-Whitney U检验分析各个Sigma因子在不同异烟肼耐药表型菌株中的表达差异。结果sigA、sigC、sigF、sigG、sigH、sigI、sigJ、sigK、sigL的基因表达量在单耐异烟肼组中的表达量高于全敏组,差异有统计学意义(Z=4.368、5.701、6.865、4.021、5.126、2.670、5.983、4.701、5.490,P均<0.001)。多耐药组中sigA、sigC、sigE、sigF、sigG、sigH、si⁃gI、sigJ、sigK、sigL的基因表达量显著高于全敏组,差异有统计学意义(Z=-5.017、-4.670、-4.667、-5.456、-4.083、-5.393、-4.712、-6.971、-8.206、-5.211,P均<0.001)。耐多药组中sigC、sigD、sigE、sigF、sigG、sigH、sigI、sigJ、sigK、sigL的基因表达量显著高于全敏组,差异有统计学意义(Z=-5.537、-4.003、-5.216、-7.328、-7.730、-5.658、-4.440、-6.036、-4.862、-4.312,P均<0.001)。sigB、sigF、sigG在异烟肼单耐药、异烟肼耐多药和异烟肼多耐药3组之间的基因表达量差异显著,有统计学意义(Z=10.139、7.735、14.532,P均<0.001)。sigF、sigG、sigI、sigJ、sigL在异烟肼耐药组中的高表达率显著高于敏感组,差异有统计学意义(χ^(2)=17.410、45.673、57.661、42.896、26.363,P均<0.001)。结论sigF、sigObjective To investigate whether the expression regulation of regulatory gene Sigma factors(sigA-sigM)is related to isoniazid resistance phenotype in isoniazid-resistant Mycobacterium tuberculosis(MTB)caused by katG mutations,and to provide reference for the study of the molecular mechanism of isoniazid resistance.Methods A total of 90 strains were collected from the patients undergoing first-line treatment at the Tianjin Tuberculosis Control Center during drug resistance testing from 2020 to 2022,of which 30 strains were sensitive strains without katG mutation,and 65 strains were isoniazidresistant strains caused by katG mutation,including 11 strains resistant only to isoniazid,24 strains resistant to both isoniazid and streptomycin,and 30 strains multidrug-resistant to isoniazid,streptomycin,and rifampicin.After the strains were collected on the isoniazid drug-containing medium,the RNA of the strains was extracted,and the relative expression levels of Sigma factors were detected by reverse transcription-polymerase chain reaction(RT-PCR).The expression differences of Sigma factors in different isoniazid drug-resistant phenotypes were analyzed by the Mann-Whitney U test.Results The gene expression levels of sigA,sigC,sigF,sigG,sigH,sigI,sigJ,sigK,sigL in isoniazid mono-resistant group were significantly higher than those in pan-isoniazid-sensitive group(Z=4.368,5.701,6.865,4.021,5.126,2.670,5.983,4.701,5.490,P all<0.001).The gene expression levels of sigA,sigC,sigF,sigG,sigH,sigI,sigJ,sigK,sigL in poly-resistance group were significantly higher than those in pan-sensitivity group(Z=-5.017,-4.670,-4.667,-5.456,-4.083,-5.393,-4.712,-6.971,-8.206,-5.211,P all<0.001).In multidrug-resistant(MDR)group,the gene expression levels of sigC,sigD,sigE,sigF,sigG,sigH,sigI,sigJ,sigK,sigL were significantly higher than those in pan-isoniazid-sensitive group(Z=-5.537,-4.003,-5.216,-7.328,-7.730,-5.658,-4.440,-6.036,-4.862,-4.312,P all<0.001).The expression levels of sigB,sigF,sigG showed statistically significant differences in

关 键 词：Sigma因子 耐药 结核 表达 

分 类 号：R378.91[医药卫生—病原生物学]