粉防己碱与多西环素联用对耐药沙门氏菌的抗菌增敏作用及机制研究  

Antimicrobial Sensitization and Mechanism of Combination of Tetrandrine and Doxycycline on Drug Resistant Salmonella

作　　者：易开放 张俊锴 刘佩仪 韩荣嘉 苗青青 潘玉善[1] 赵金凤[1] 翟亚军 胡功政[1] YI Kai-fang;ZHANG Jun-kai;LIU Pei-yi;HAN Rong-jia;MIAO Qing-qing;PAN Yu-shan;ZHAO Jin-feng;ZHAI Ya-jun;HU Gong-zheng(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou,Henan,450046,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046

出　　处：《动物医学进展》2023年第1期65-69,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32072913)。

摘　　要：旨在探讨粉防己碱与多西环素联合应用对沙门氏菌的体外协同抗菌及其作用机制,为临床联合用药控制耐药性沙门氏菌感染提供理论依据。采用微量肉汤稀释法分别测定粉防己碱与多西环素对沙门氏菌的最小抑菌浓度(minimum inhibitoty concentration,MIC),棋盘法测定联合用药后的部分抑菌浓度指数(FICI),时间-杀菌曲线验证联合用药的抑菌效果,应用荧光染料溴化乙锭(EtBr)作为外排泵的作用底物,示踪联合用药后沙门氏菌外排泵的活性。结果表明,粉防己碱与多西环素对沙门氏菌的MIC分别为4μg/mL~64μg/mL、320μg/mL~640μg/mL,两药联用的平均FICI为0.46,协同作用占84.62%,相加作用占15.38%;时间-杀菌曲线也证明两药联用具有良好的协同或相加作用;EtBr蓄积试验表明,粉防己碱与多西环素联合组的荧光强度极显著高于多西环素单药组和空白对照组(P<0.01),表明粉防己碱可显著抑制外排泵的活性。粉防己碱与多西环素联用呈协同或相加作用,粉防己碱可明显增强多西环素对沙门氏菌的抗菌活性,其作用机制之一是通过抑制外排泵活性,增强细菌对多西环素的敏感性。This experiment was conducted to investigate the synergistic antibacterial effect of tetrandrine and doxycycline on Salmonella in vitro and its mechanism,and to provide theoretical basis for clinical combined drug control of drug-resistant Salmonella infection.The minimum inhibitory concentrations(MIC)of tetrandrine and doxycycline against Salmonella were determined by broth dilution method,and then the partial inhibitory concentration index(FICI)was determined by checkerboard method.The bacteriostatic effect of the combination was verified by the time-kill curve method.Fluorescent dye ethidium bromide(EtBr)was used as the substrate of the efflux pump to trace the activity of Salmonella efflux pump.The results showed that the MIC of tetrandrine and doxycycline against Salmonella were 4-64μg/mL and 320-640μg/mL,respectively.The average FICI of the two drugs combined was 0.47,accounting for 84.62%synergistic effect and 15.38%additive effect.The time-kill curve also proved that the combination of tetrandrine and doxycycline had a good synergistic or additive effect.EtBr accumulation test showed that the fluorescence intensity of tetrandrine combined with doxycycline was significantly higher than that of doxycycline single drug and control group(P<0.01),indicating that tetrandrine could significantly inhibit the activity of the efflux pump.This experiment proved that the combination of tetrandrine and doxycycline had synergistic or additive effect,and tetrandrine could significantly enhance the antibacterial activity of doxycycline against Salmonella.One of the mechanisms was to enhance the sensitivity of bacteria to drugs by inhibiting the activity of efflux pump.

关 键 词：粉防己碱 多西环素 协同作用 外排泵活性 沙门氏菌 

分 类 号：S852.612[农业科学—基础兽医学] S853.74[农业科学—兽医学]