RND外排泵对多重耐药鲍曼不动杆菌AYE菌株生物被膜形成的影响  

Effect of RND efflux pump on biofilm formation of multidrug-resistant Acinetobacter baumannii AYE strain

作　　者：王清会 蔺飞 刘献清 凌保东[1,2] WANG Qing-hui;LIN Fei;LIU Xian-qing;LING Bao-dong(Research Institute of Structure-specific Small Molecule Drugs,Chengdu Medical College,Key Laboratory of Universities of Sichuan Province,Chengdu 610500,China;School of Pharmacy,Chengdu Medical College,Chengdu 610500,China;Department of Pharmacy,Chengdu Jinniu District People’s Hospital,Chengdu 610500,China;Department of Pharmacy,First Affiliated Hospital,Chengdu Medical College Clinical School,Chengdu 610500,China;Clinical Pharmacy Office,Guangyuan City Hospital of Traditional Chinese Medicine,Guangyuan 628099,China)

机构地区：[1]成都医学院结构特异性小分子药物研究四川省高校重点实验室,四川成都610500 [2]成都医学院药学院,四川成都610500 [3]成都市金牛区人民医院西药剂科,四川成都610500 [4]成都医学院临床医学院第一附属医院药学部,四川成都610500 [5]广元市中医院临床药学办,四川广元628099

出　　处：《中国感染控制杂志》2021年第10期876-880,共5页Chinese Journal of Infection Control

基　　金：国家自然科学基金(81373454)。

摘　　要：目的探讨多重耐药鲍曼不动杆菌AYE菌株RND外排泵基因影响生物被膜形成的作用机制。方法采用无标记基因敲除方法分别敲除AYE菌株的RND外排泵基因(ade B、ade FGH、ade IJK),采用实时荧光定量聚合酶链式反应(qRT-PCR)验证菌株外排泵基因敲除情况,通过结晶紫染色法检测菌株基因敲除前后生物被膜形成情况,比较4种外排泵抑制剂(PAβN、奥美拉唑、维拉帕米、CCCP)对AYE菌株生物被膜形成的影响。结果qRT-PCR结果显示,AYE菌株RND外排泵基因(ade B、ade FGH、ade IJK基因)成功敲除,获得AYE△ade B、AYE△ade FGH、AYE△ade IJK菌株,生物被膜形成量分别为1.59±0.06、2.15±0.19、1.91±0.02,其中ade B和ade IJK基因敲除后菌株生物被膜形成量低于敲除前(AYE:2.31±0.01),差异均有统计学意义(均P<0.05)。在PAβN、奥美拉唑、维拉帕米、CCCP作用下,鲍曼不动杆菌标准菌株AYE的生物被膜形成量分别为2.14±0.03、2.24±0.02、1.93±0.05、2.09±0.04,均低于对照组(AYE:2.31±0.01),差异均有统计学意义(均P<0.05),其中PAβN形成量最少。结论外排泵基因ade B、ade FGH、ade IJK对多重耐药鲍曼不动杆菌AYE生物被膜的形成有重要作用,外排泵抑制剂PAβN、奥美拉唑、维拉帕米、CCCP对鲍曼不动杆菌AYE菌株生物被膜的形成有抑制作用。Objective To explore the mechanism of resistance-nodulation-cell division(RND)efflux pump genes of multidrug-resistant Acinetobacter baumannii(MDR-AB)AYE strain in influencing biofilm formation.Methods RND efflux pump genes(ade B,ade FGH,ade IJK)of Acinetobacter baumannii AYE strain were knocked out by marker-free gene knockout method,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to verify the gene knockout of strain efflux pump genes,biofilm formation before and after gene knockout was detected by crystal violet staining,effect of 4 efflux pump inhibitors(PAβN,omeprazole,verapamil and CCCP)on biofilm formation of AYE strain was compared.Results qRT-PCR showed that RND efflux pump genes(ade B,ade FGH,ade IJK genes)of AYE strain were successfully knocked out,AYE△ade B,AYE△ade FGH,and AYE△ade IJK strains were obtained,the amount of biofilm formation was 1.59±0.06,2.15±0.19 and 1.91±0.02 respectively,the amount of biofilm formation after ade B and ade IJK gene knockout was lower than that before knockout(AYE:2.31±0.01),difference were all statistically significant(all P<0.05).Under the effect of PAβN,omeprazole,verapamil and CCCP,the amount of biofilm formation of standard strain AYE of Acinetobacter baumannii were 2.14±0.03,2.24±0.02,1.93±0.05 and 2.09±0.04 respectively,which were all lower than those in the control group(AYE:2.31±0.01),difference was statistically significant(all P<0.05),PAβN formation was the least.Conclusion Efflux pump genes ade B,ade FGH and ade IJK play important role in the formation of biofilm of MDR-AB AYE,efflux pump inhibitors PAβN,omeprazole,verapamil and CCCP have an inhibitory effect on biofilm formation of Acinetobacter baumannii AYE strain.

关 键 词：鲍曼不动杆菌 生物被膜 RND外排泵 外排泵抑制剂 

分 类 号：R181.32[医药卫生—流行病学] R446.5[医药卫生—公共卫生与预防医学]