摘要
目的盐酸小檗碱(Ber)联合喹诺酮(FQN_(S))对产超广谱β内酰胺酶(ESBLs)肺炎克雷伯菌(KPn)的体外抑菌作用。方法采用微量稀释法测定Ber或FQN_(S):环丙沙星(CIP)、左氧氟沙星(LVX)及加替沙星(GAT)单药对KPn的最低抑菌浓度(MIC),棋盘法测定两药联用的MIC并计算FIC指数;选取KPn 11、KPn 15菌株测定单用Ber或联用CIP、LVX、GAT的杀菌曲线并绘制图形;RT-PCR测定外排泵基因acrA、acrB和tolC的表达量。结果与单用CIP、LVX、GAT相比,联合Ber后,KPn对CIP、LVX、GAT的MIC值下降率100%,均能下降1倍以上(P<0.05),其中联用后GAT的MIC值下降2倍达50%,CIP、LVX与Ber联用主要呈相加作用,分别为75%和70%,GAT与Ber联用主要呈协同作用50%;杀菌曲线测定中,CIP、LVX与Ber联用及单用Ber曲线均有所下降,但差异较小,能抑制细菌生长但效果不明显(P>0.05),但GAT与Ber联用曲线明显下降,抗菌活性好,能有效抑制细菌生长(P<0.05);单用Ber与空白组相比acrA、acrB和tolC表达量有3~5倍升高(P<0.05),CIP、LVX及GAT与Ber联用与空白组相比表达量明显升高,15~19倍升高(P<0.05),但CIP、LVX及GAT与Ber联用,组间表达量无明显差异(P>0.05)。结论本实验证明了Ber与FQN_(S)联用能有效抑制产ESBLs KPn的生长,增加FQN_(S)对KPn的抗菌作用,GAT与Ber联用时抗菌作用最为明显。
Objective In vitro antibacterial effect of berberine on extended-spectrumβ-lactamase-producing Klebsiella pneumoniae combing with quinolone(FQN_(S)).Methods The microdilution method was used to determine the Ber or FQN_(S):the minimum inhibitory concentration(MIC)of ciprofloxacin(CIP),levofloxacin(LVX)and gatifloxacin(GAT)on KPn was determined by the checkerboard method.KPn 11 and KPn 15 strains were selected to determine the sterilization curve of Ber alone or in combination with CIP,LVX,GAT and graphs;RT-PCR was used to determine the expression of efflux pump genes acrA,acrB and tolC.Results Compared with CIP,LVX,and GAT alone,the MIC value of KPn on CIP,LVX,and GAT decreased by 100%after combined with Ber,and all of them decreased by more than 1 times(P<0.05).Among them,the MIC of GAT after combined use The value decreased by 2 times up to 50%.The combination of CIP,LVX and Ber mainly showed an additive effect,75%and 70%,respectively.The combination of GAT and Ber mainly showed a synergistic effect of 50%;in the determination of the sterilization curve,CIP,LVX and Both Ber combined and single-use Ber curves have decreased,but the difference is small.It can inhibit the growth of bacteria but the effect is not obvious(P>0.05),but the combined use of GAT and Ber has a significant decrease in antibacterial activity and can effectively inhibit bacteria growth(P<0.05);the expression of acrA,acrB,and tolC increased by 3-5 times compared with the blank group with Ber alone(P<0.05),and the expression of CIP,LVX and GAT in combination with Ber was compared with the blank group The amount was significantly increased,15~19 times higher(P<0.05),but CIP,LVX and GAT combined with Ber,there was no significant difference in expression between the groups(P>0.05).Conclusion This experiment proved that the combination of Ber and FQN_(S) can effectively inhibit the growth of ESBLs-producing KPn,and increase the antibacterial effect of FQN_(S) on KPn.The antibacterial effect of GAT and Ber is the most obvious.
作者
周晓元
罗润齐
王俊洁
张宿荣
许晓明
杨洋
叶晓光
ZHOU Xiaoyuan;LUO Runqi;WANG Junjie;ZHANG Surong;XU Xiaoming;YANG Yang;YE Xiaoguang(Department of Infectious Diseases,Second Affiliated Hospital,Guangzhou Medical University,Guangzhou 510260,Guangdong Province,China)
出处
《现代医院》
2021年第10期1613-1618,共6页
Modern Hospitals
基金
广州市卫生和计划生育科技项目西医类一般引导项目(20181A011075)
广州市卫生和计划生育科技项目西医类一般引导项目(20181A010056)
广州医科大学青年科研基金资助项目(52010202-0940)。
