黄连素对耐多黏菌素大肠杆菌的耐药性逆转和mcr-1基因消减作用  

Reversal of berberine resistance to polymyxin-resistant Escherichia coli and reduction of mcr-1 gene

作　　者：王新兴 刘有旺 张轩 王嘉慧 常维山[2] 王巧云 WANG Xinxing;LIU Youwang;ZHANG Xuan;WANG Jiahui;CHANG Weishan;WANG Qiaoyun(School of Public Health and Health Management,Shandong First Medical University,Shandong Academy of Medical Sciences,Shandong,Tai’an 271016,China;College of Animal Science and Technology,Shandong Agricultural University,Shandong,Tai’an 271018,China;Department of Hepatology,Tai’an Medical District,the 960th Hospital of Chinese People’s Liberation Army,Shandong,Tai’an 271000,China)

机构地区：[1]山东第一医科大学山东省医学科学院公共卫生与健康管理学院,山东泰安271016 [2]山东农业大学动物科技学院,山东泰安271018 [3]解放军960医院泰安医疗区肝病科,山东泰安271000

出　　处：《中国医药科学》2022年第1期129-132,共4页China Medicine And Pharmacy

摘　　要：目的建立黄连素逆转mcr-1阳性大肠杆菌耐药性的方法,为中药抗多重耐药菌感染提供新线索。方法通过肉汤稀释法分别测定黄连素和氯丙嗪对大肠杆菌EMCRR45的最低抑菌浓度,并以此浓度进行质粒消除实验。利用棋盘培养实验确认耐药质粒的消除,计算消除率,使用SPSS 17.0软件与对照组氯丙嗪组进行消除率比较。利用K-B法或微量肉汤稀释法检测质粒消除前后耐药性的变化。采用荧光定量PCR检测耐药质粒消除前后细菌中mcr-1含量的变化。结果黄连素和氯丙嗪对大肠杆菌EMCRR45的最低抑菌浓度分别为1.25 mg/ml和68μg/ml,72 h的质粒消除率分别为26.9%和6.0%,差异有统计学意义(P=0.001)。质粒消除后,耐受抗生素种类从8种下降至4种,同时逆转了对多黏菌素的耐药性。荧光定量PCR显示,质粒消除后mcr-1的CT值从28.55变为34.50。结论黄连素可以消减大肠杆菌mcr-1基因,并逆转细菌对多黏菌素的耐药性。Objective To establish a method for berberine to reverse the drug resistance of mcr-1 positive Escherichia coli,and to provide a new clue for Chinese medicine to resist multi-drug resistant bacteria infection.Methods The minimum inhibitory concentrations(MICs)of berberine and chlorpromazine against Escherichia coli EMCRR45 were determined by broth dilution method,and plasmid elimination experiments were carried out at these concentrations.The elimination of drug-resistant plasmid was confirmed by checkerboard culture experiment,and the elimination rate was calculated.SPSS 17.0 software was used to compare the elimination rate of the berberine group with the chlorpromazine group(the control group).The changes of drug resistance before and after plasmid elimination were detected by K-B method or broth dilution method.The changes of mcr-1 content in bacteria before and after elimination of drug-resistant plasmid were detected by fluorescence quantitative-polymerase chain reaction(PCR).Results The MIC of berberine and chlorpromazine against Escherichia coli EMCRR45 was 1.25 mg/ml and 68μg/ml,respectively,and the elimination rate of plasmids in 72 hours was 26.9%and 6.0%,respectively,with significant difference(P=0.001).After plasmid elimination,the type of resistant antibiotics decreased from 8 to 4,and the resistance to polymyxin was reversed.The fluorescence quantitative PCR showed that the CT value of mcr-1 changed from 28.55 to 34.50 after plasmid elimination.Conclusion Berberine can reduce mcr-1 gene of Escherichia coli and reverse the drug resistance of the bacteria to polymyxin.

关 键 词：大肠杆菌 质粒 黄连素 mcr-1 

分 类 号：R446.5[医药卫生—诊断学]