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目的探讨类泛素-蛋白酶体系统对结核分枝杆菌单纯利福平耐药性的影响。方法采用刃天青显色法检测利福平对结核分枝杆菌的最低抑菌浓度(minimum inhibitory concentration,MIC),比较分析结核分枝杆菌Pup、Dop、PafA、Mpa基因的过表达或缺失突变对结核分枝杆菌利福平MIC的差异;检测分别加入羰基氰氯苯腙、利血平、维拉帕米和氯丙嗪4种外排泵抑制剂前后各菌株对利福平MIC的影响。结果结核分枝杆菌Pup、Dop、PafA和Mpa基因的过表达均能增强单纯耐利福平结核分枝杆菌对利福平的耐药性,而Pup、Mpa、Dop和PafA基因的缺失均能显著降低单纯耐利福平结核分枝杆菌对利福平的耐药性,P值均<0.05。4种药物外排泵抑制剂能不同程度的降低各过表达菌株对利福平的MIC,P值均<0.05,并且,类泛素-蛋白酶体系统与外排泵抑制剂之间存在一定交互作用。结论类泛素-蛋白酶体系统对结核分枝杆菌单纯利福平耐药性的产生有影响;类泛素-蛋白酶体系统可能通过调控外排相关通路蛋白来影响结核分枝杆菌单纯利福平耐药性的产生。
Objective To investigate the effects of ubiquitin-proteasome system on the drug resistance of Mycobacterium tuberculosis to rifampicin. Methods The minimum inhibitory concentration (MIC) of rifampicin against Mycobacterium tuberculosis was determined by the resazurin colorimetric method. The over-expression or deletion mutations of Pup, Dop, PafA and Mpa genes were compared and analyzed. On MICs of rifampicin for Mycobacterium tuberculosis; the effect of each strain on the MIC of rifampicin before and after the addition of 4 efflux pump inhibitors of carbonyl cyanamide, reserpine, verapamil and chlorpromazine respectively . Results Overexpression of Mycobacterium tuberculosis Pup, Dop, PafA and Mpa enhanced the resistance of rifampicin-resistant Mycobacterium tuberculosis to rifampin, while the deletion of Pup, Mpa, Dop and PafA genes Significantly reduce the resistance of rifampicin-resistant Mycobacterium tuberculosis to rifampin, all P <0.05.4 Inhibitors of drug efflux pump can reduce the MIC, P <0.05, and there was some interaction between the ubiquitin-proteasome system and efflux pump inhibitors. Conclusion The ubiquitin-proteasome system has an influence on the drug resistance of Mycobacterium tuberculosis, and the ubiquitin-proteasome system may influence the Mycobacterium tuberculosis rifampicin by regulating the efflux-related pathway proteins Drug resistance.