青蒿琥酯联合头孢菌素类药物对多重耐药肺炎克雷伯菌抗菌活性的分析  

Analysis on Antibacterial Activity of Artesunate in Combination with Cephalosporins to Multidrug-resistant Klebsiella pneumoniae

作　　者：晏国威 马小华[2] 潘建华[2] 喻容[2] 石国民[2] 向延根[2] Yan Guowei;Ma Xiaohua;Pan Jianhua;Yu Rong;Shi Guomin;Xiang Yangen(Changsha Central Hospital Affiliated to University of South China,Changsha,410004;Clinical Laboratory,Changsha Central Hospital,Changsha,410004)

机构地区：[1]南华大学附属长沙中心医院,长沙410004 [2]长沙市中心医院检验科,长沙410004

出　　处：《基因组学与应用生物学》2021年第3期1391-1399,共9页Genomics and Applied Biology

基　　金：湖南省卫生和计生委员会科研课题(B2016156)资助。

摘　　要：为探讨青蒿琥酯(artesunate,ASN)单独或联合头孢菌素类药物对多重耐药(multidrug-resistant,MDR)肺炎克雷伯菌(Klebsiella pneumoniae)的抗菌活性。本研究收集2018年长沙市中心医院临床分离的多重耐药肺炎克雷伯菌菌株32株。分为单独用药组:青蒿琥酯(ASN)组、头孢曲松钠(CRO)/头孢他啶(CAZ)组和联合应用组。采用细菌动态生长曲线法观察青蒿琥酯联合头孢菌素类药物对多重耐药肺炎克雷伯菌的生长影响;采用微量肉汤稀释法检测青蒿琥酯单独或联合头孢菌素类药物对多重耐药肺炎克雷伯菌的抗菌作用;采用RT-PCR技术检测外排泵基因AcrB mRNA以及其调控基因MarA和AcrR mRNA的表达水平。结果显示,终浓度为2048μg/mL ASN联合头孢曲松钠、头孢他啶明显地抑制了多重耐药肺炎克雷伯菌的生长,曲线走向平缓;终浓度为1024μg/mL ASN联合头孢曲松钠、头孢他啶对多重耐药肺炎克雷伯菌的MIC50与单独用药组比较显著减低,均有统计学意义(K=68.07,P<0.05;K=62.34,P<0.05)。1024μg/mL ASN联合头孢曲松钠、头孢他啶均以相加作用为主;2048μg/mL青蒿琥酯和头孢曲松钠、头孢他啶联用时,可以抑制外排泵基因AcrB的表达,2048μg/mL青蒿琥酯联合512μg/mL头孢曲松钠明显抑制正向调控子MarA的表达,而对于负性调控子AcrR的表达没有明显影响。青蒿琥酯能显著改善多重耐药肺炎克雷伯菌对头孢曲松钠以及头孢他啶的敏感性,其机制可能与下调外排泵基因AcrB的表达有关。To investigate the antibacterial activity of artesunate(ASN)alone or in combination with cephalosporins on multidrug-resistant(MDR)Klebsiella pneumoniae,a total of 32 strains of multidrug-resistant K.pneumoniae isolated from the Changsha Central Hospital were collected in 2018 and divided into separate drug groups:artesunate(ASN)groups,ceftriaxone sodium(CRO)/ceftazidime(CAZ)groups and combined application groups.The effect of artesunate combined with cephalosporins on the growth of multidrug-resistant K.pneumoniae was observed by the method of bacterial dynamic growth curve.The antibacterial effect of artesunate alone or in combination with cephalosporins on multidrug-resistant K.pneumoniae was determined by the method of micro-broth dilu tion.The expression level of AcrB mRNA and its regulatory genes MarA and AcrR mRNA were detected by RT-PCR.The results showed that the final concentration of 2048μg/mL ASN combined with ceftriaxone sodium or ceftazidime significantly inhibited the growth of multi-drug resistant K.pneumoniae,and the curve was gentle;the final concentration of 1024μg/mL ASN combined with ceftriaxone sodium or ceftazidime was significantly lower the MIC50 of multidrug-resistant K.pneumoniae than the drug-administered groups(K=68.07,P<0.05;K=62.34,P<0.05).1024μg/mL ASN combined with ceftriaxone sodium or ceftazidime were mainly additive;when 2048μg/mL artesunate was combined with ceftriaxone sodium or ceftazidime,the expression of AcrB in the efflux pump gene was inhibited.when 2048μg/mL artesunate combined with 512μg/mL ceftriaxone sodium significantly inhibited the expression of the positive regulator MarA,but had no significant effect on the expression of the negative regulator AcrR.Artesunate significantly improved the sensitivity of multidrug-resistant K.pneumoniae to ceftriaxone sodium and ceftazidime,and its mechanism may be related to the down-regulation of the expression of the efflux pump gene AcrB.

关 键 词：青蒿琥酯 多重耐药肺炎克雷伯菌 头孢曲松钠 头孢他啶 抗菌作用 

分 类 号：R965[医药卫生—药理学]