CAJ | 학술논문

结核杆菌Pup-蛋白酶体系统与耐药结核杆菌耐药性的相关性研究
결핵간균Pup-단백매체계통여내약결핵간균내약성적상관성연구
Correlation study between prokaryotic ubiquitin-like protein (Pup)-proteasome and drug resistance of clinical isolates of Mycobacterium tuberculosis

万方数据

中国免疫学杂志中國免疫學雜誌 중국면역학잡지
CHINESE JOURNAL OF IMMUNOLOGY
2014年 11期 1441-1447,1451 ,共8页

何丽%雷英%吴芳%章乐%吴江东%曹旭东%朱彬%邬博%李瑞山%张玉清%王钊%庄睿%李文娟%梁晨%樊超%张万江

하려%뢰영%오방%장악%오강동%조욱동%주빈%오박%리서산%장옥청%왕쇠%장예%리문연%량신%번초%장만강

结核杆菌%耐药性%Pup-蛋白酶体系统結覈桿菌%耐藥性%Pup-蛋白酶體繫統
결핵간균%내약성%Pup-단백매체계통
Mycobacterium tuberculosis%Drug-resistantance%Prokaryotic ubiquitin-like protein ( Pup)-proteasome

目的：检测不同耐药的结核杆菌临床分离株原核类泛素蛋白-蛋白酶体系统（以下简称：Pup-蛋白酶体系统）相关的Pup、Dop、PafA、Mpa基因的表达水平，探讨研究结核杆菌Pup-蛋白酶体系统与新疆地区广泛流行的耐药结核杆菌临床分离株耐药性的相关性。方法：以培养对数生长期的新疆地区广泛流行的耐药结核杆菌临床分离株为研究对象，分别提取结核杆菌临床分离的药物敏感菌株、单纯耐异烟肼的结核杆菌临床分离株（ INH-MTB）、单纯耐利福平的结核杆菌临床分离株（RFP-MTB）、单纯耐链霉素的结核杆菌临床分离株（SM-MTB）、单纯耐乙胺丁醇的结核杆菌临床分离株（EB-MTB）、耐多药结核杆菌临床分离株（ MDR）等各组菌株的总RNA，经纯度鉴定后，利用SYBR GreenⅠ实时荧光定量PCR方法，检测各组结核杆菌菌株的Pup、Dop、PafA、Mpa基因的表达水平，分析各组耐药结核杆菌临床分离株Pup-蛋白酶体系统相关的Pup、Dop、PafA、Mpa基因的表达与新疆地区广泛流行的耐药结核杆菌临床分离株耐药性的相关性。结果：与药物敏感菌株相比，Pup、Mpa基因在单一耐药菌株和耐多药菌株中表达有不同程度的下调，Dop、PafA基因在单一耐药菌株和耐多药菌株中表达有不同程度的上调，差异有统计学意义（P＜0.05）。与MDR相比，Pup、Dop、Mpa基因在单一耐药菌株中表达有不同程度的上调，PafA基因在单一耐药菌株中表达有不同程度的下调，差异有统计学意义（ P＜0.05）。结论：在结核杆菌临床分离的药物敏感菌株、INH-MTB组、RFP-MTB组、SM-MTB组、EB-MTB组和MDR组中，结核杆菌Pup、Dop、PafA、Mpa基因的表达差异，与新疆地区广泛流行的耐药结核杆菌临床分离株耐药性有相关性。
목적：검측불동내약적결핵간균림상분리주원핵류범소단백-단백매체계통（이하간칭：Pup-단백매체계통）상관적Pup、Dop、PafA、Mpa기인적표체수평，탐토연구결핵간균Pup-단백매체계통여신강지구엄범류행적내약결핵간균림상분리주내약성적상관성。방법：이배양대수생장기적신강지구엄범류행적내약결핵간균림상분리주위연구대상，분별제취결핵간균림상분리적약물민감균주、단순내이연정적결핵간균림상분리주（ INH-MTB）、단순내리복평적결핵간균림상분리주（RFP-MTB）、단순내련매소적결핵간균림상분리주（SM-MTB）、단순내을알정순적결핵간균림상분리주（EB-MTB）、내다약결핵간균림상분리주（ MDR）등각조균주적총RNA，경순도감정후，이용SYBR GreenⅠ실시형광정량PCR방법，검측각조결핵간균균주적Pup、Dop、PafA、Mpa기인적표체수평，분석각조내약결핵간균림상분리주Pup-단백매체계통상관적Pup、Dop、PafA、Mpa기인적표체여신강지구엄범류행적내약결핵간균림상분리주내약성적상관성。결과：여약물민감균주상비，Pup、Mpa기인재단일내약균주화내다약균주중표체유불동정도적하조，Dop、PafA기인재단일내약균주화내다약균주중표체유불동정도적상조，차이유통계학의의（P＜0.05）。여MDR상비，Pup、Dop、Mpa기인재단일내약균주중표체유불동정도적상조，PafA기인재단일내약균주중표체유불동정도적하조，차이유통계학의의（ P＜0.05）。결론：재결핵간균림상분리적약물민감균주、INH-MTB조、RFP-MTB조、SM-MTB조、EB-MTB조화MDR조중，결핵간균Pup、Dop、PafA、Mpa기인적표체차이，여신강지구엄범류행적내약결핵간균림상분리주내약성유상관성。
Objective:To study the correlation different drug-resistant mycobacterium tuberculosis clinical isolates of prokaryotic ubiquitin-like protein ( Pup )-proteasome of Pup, Dop, PafA, Mpa gene expression level and mycobacterium tuberculosis Pup-proteasome system with Xinjiang region widely popular drug-resistant mycobacterium tuberculosis in clinical isolates resistance.Methods:Total RNA of Mtb was extracted from cultured Mtb during the logarithmic phase in drug-susceptible strains in Xinjiang region,the clinical strains drug sensitive to INH,RFP,SM and EB respectively,and multidrug-resistant(MDR) strains.And then the purity of total RNA was identified.The expressing of Pup-proteasome relevant gene( Pup,Dop,Mpa,PafA) were quantified using SYBR GreenⅠqRT-PCR which aimed at finding the correlation between Mtb Pup-proteasome system and drug resistance of Mtb clinical isolates widespread in Xingjiang region by analyzing the expression of Pup, Dop, PafA, Mpa gene among different isolates.Results:Compared with the drug-senstive clinical isolates, mRNA expression level of Pup, Mpa gene was down-regulated in resistant M.tuberculosis clinical isolates INH ( INH-MTB) ,RFP ( RFP-MTB) ,SM ( SM-MTB) and EB ( EB-MTB) ,mRNA expression levels of genes in Dop and PafA was higher in resistant M.tuberculosis clinical isolates,the difference was statistically significant(P<0.05).Compared with MDR strain, the expression of Pup, Dop, Mpa gene were up-regulated different in the resistant M.tuberculosis clinical isolates isolates ,the expression of PafA gene was down-regulated different,the difference was statistically significant( P<0.05).Conclusion:The differentially expressed gene of Pup、Dop、PafA、Mpa gene in sensitive strains,INH-MTB,RFP-MTB,SM-MTB,EB-MTB and MDR strains.The Mycobacterium tuberculosis Pup-proteasome system.Therefore,the Pup-proteasome system have association with the drug resistance of Mtb strains widespread in Xinjiang region.