蟾酥醇提物对多重耐药铜绿假单胞菌头孢他啶敏感性的影响  

Effect of Ethanol Extract of Bufonis Venenum on Sensitivity of Multidrug-resistant Pseudomonas aeruginosa(MDRPA)to Ceftazidime

作　　者：周鸿威 王成祥 吕思缘 林英[3] 张悦 刘国星 李磊 王玉婷 郭雨菲 薛贝 于会勇 ZHOU Hongwei;WANG Chengxiang;LYU Siyuan;LIN Ying;ZHANG Yue;LIU Guoxing;LI Lei;WANG Yuting;GUO Yufei;XUE Bei;YU Huiyong(Respiratory Department,Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100020,China;Pulmonary Disease Department,Jiangmen Wuyi Traditional Chinese Medicine Hospital,Jiangmen 529000,China;Clinical Laboratory Department,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100010,China;Hepingli Hospital of Beijing,Beijing 100010,China)

机构地区：[1]北京中医药大学第三附属医院呼吸科,北京100020 [2]江门市五邑中医院肺病科,江门529000 [3]北京中医药大学东直门医院检验科,北京100010 [4]北京市和平里医院,北京100010

出　　处：《世界中医药》2024年第23期3592-3596,共5页World Chinese Medicine

基　　金：国家自然科学基金项目(82074389);北京市自然科学基金项目(7202127);北京中医药大学第三附属医院2019年度科研孵化项目(2019kyfh-10);北京市中医药大学第三附属医院2022年度院级培育项目(BZYSY-2022-PYMS-16)。

摘　　要：目的:探讨蟾酥醇提物通过铜绿假单胞菌主动外排系统(MexABOprM)对多重耐药铜绿假单胞菌(MDRPA)头孢他啶敏感性的影响。方法:筛选MexAB-OprM阳性的铜绿假单胞菌菌株,通过肉汤稀释法建立MDRPA体外抑菌实验模型,测定蟾酥醇提物及其联合头孢他啶时的最小抑菌浓度;蟾酥醇提物干预前后用实时荧光反转录PCR检测MDRPA铜绿假单胞菌主动外排系统MexAB-OprM中mexB、mexR、oprM基因mRNA表达量。结果:蟾酥醇提物对MDRPA的最小抑菌浓度值是6.25 mg/mL。蟾酥醇提物联合头孢他啶的联合指数低于1,两药联合应用时存在相加作用。随蟾酥醇提物浓度升高,MDRPA外排泵MexAB-OprM的mexB、oprM相对表达量降低(P<0.05),mexR相对表达量升高(P<0.05)。结论:蟾酥醇提物通过抑制MexABOprM外排泵的表达提高MDRPA对头孢他啶的敏感性。Objective:To explore the effect of Bufonis Venenum ethanol extract on the sensitivity of multidrug-resistant Pseudomonas aeruginosa(MDRPA)to ceftazidime through the MexAB-OprM efflux pump system.Methods:P.aeruginosa strains positive for the MexAB-OprM system were screened.An in vitro antibacterial model for MDRPA was established using the broth dilution method,and the minimum inhibitory concentration(MIC)of the Bufonis Venenum ethanol extract and its combination with ceftazidime was determined.The mRNA expression levels of mexB,mexR,and oprM genes in the MexAB-OprM efflux pump system were detected by real-time fluorescence reverse transcription PCR before and after intervention with the Bufonis Venenum ethanol extract.Results:The MIC value of the Bufonis Venenum ethanol extract against MDRPA was 6.25 mg/mL.The combination index of Bufonis Venenum ethanol extract with ceftazidime was less than 1,indicating a synergistic effect between the two drugs.As the concentration of the Bufonis Venenum ethanol extract increased,the relative expression levels of mexB and oprM in the MexAB-OprM efflux pump decreased(P<0.05),while the relative expression of mexR increased(P<0.05).Conclusion:The Bufonis Venenum ethanol extract enhances the sensitivity of MDRPA to ceftazidime by inhibiting the expression of the MexAB-OprM efflux pump.

关 键 词：蟾酥醇提物 多重耐药铜绿假单胞菌 体外抑菌 肉汤稀释法 实时荧光反转录聚合酶链反应 铜绿假单胞菌主动外排系统 抗耐药作用 逆转外排泵机制 

分 类 号：R285.5[医药卫生—中药学]