碳青霉烯类药物对铜绿假单胞菌生物膜形成及其基因表达的影响  

Effects of carbapenems on the biofilm formation and gene expression of Pseudomonas aeruginosa

作　　者：周蓓蓓[1] 王冲 杜佳 周翠[1] 刘海洋[1] 张艺之 周铁丽[1] Zhou Bei-bei;Wang Chong;Du Jia;Zhou Cui;Liu Hai-yang;Zhang Yi-zhi;Zhou Tie-li(Laboratory Medicine Center,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325035;Clinical Laboratory,Zhejiang Cancer Hospital,Hangzhou 310000)

机构地区：[1]温州医科大学附属第一医院医学检验中心,温州325035 [2]浙江省肿瘤医院检验科,杭州310000

出　　处：《中国抗生素杂志》2019年第12期1394-1399,共6页Chinese Journal of Antibiotics

摘　　要：目的研究碳青霉烯类药物对铜绿假单胞菌生物膜形成和基因表达的影响,为治疗铜绿假单胞菌引起的感染提供分子基础和理论依据。方法选取3株临床分离的对碳青霉烯类药物敏感,但对其他常用抗菌药物耐药的铜绿假单胞菌(TL1552、TL1728和TL947),采用琼脂稀释法检测3株菌对临床常用抗菌药物的最低抑菌浓度(mininal inhibotory concentration,MIC);结晶紫染色法及扫描电子显微镜观察不同浓度碳青霉烯类药物对铜绿假单胞菌生物膜形成能力和形态的影响;通过转录组测序分析生物膜相关的差异表达基因,并用qRT-PCR进行确认。结果亚胺培南和美罗培南均能有效抑制铜绿假单胞菌生物膜的形成,但对已经形成的生物膜影响较小;扫描电子显微镜显示美罗培南处理后的生物膜细胞数量、大小和形态均发生了显著改变,继续在不含抗菌药物的培养基上孵育6h后发现生物膜细胞的整体形态基本恢复正常;转录组测序分析及qRT-PCR表明碳青霉烯类药物可导致涉及蛋白质运输、代谢、生物膜形成等相关基因的改变,并能触发生物膜细胞的应激反应。结论碳青霉烯类药物对铜绿假单胞菌的生物膜形成有抑制作用,但对已经形成的生物膜影响甚微,去除药物后生物膜细胞的形态基本得到恢复,说明铜绿假单胞菌感染后早期治疗和足疗程治疗的重要性。Objective To evaluate the effects of carbapenems on the biofilm formation and gene expression of Pseudomonas aeruginosa.Methods Three P.aeruginosa strains were included in our study and the minimal inhibitory concentrations(MICs)were tested through the agar dilution method.Crystal violet staining assay and scanning electron microscopy were performed to evaluate the effects of different concentrations of carbapenems on the cell number and morphology of P.aeruginosa biofilms.Transcriptome sequencing was carried out to select differential expression genes,which were then verified by quantitative RT-PCR(qRT-PCR).Results Three P.aeruginosa strains were resistant to many antibiotics except carbapenems.The biofilm formation decreased concurrently in all isolates when treated with imipenem and meropenem,but no significant effect was observed on the mature biofilm.However,the size,the shape and the number of biofilm cells changed significantly after the treatment with meropenem.Nevertheless,distinct recovery of the biofilm cells was observed at 6 h after removal of carbapenems.Transcriptome sequencing and quantitative RT-PCR(qRT-PCR)results suggested that carbapenems cause evident change in gene expression involved in protein transport,metabolism,and biosynthesis of membrane protein,and activate the stress response of biofilm cells.Conclusions Carbapenems have an inhibitory effect on the biofilm formation of P.aeruginosa,but have little effect on the biofilm that has formed.The morphology of biofilm got a satisfactory recovery after withdrawal of carbapenems.Therefore,this study highlights the importance of early treatment and foot treatment after the infection of P.aeruginosa.

关 键 词：铜绿假单胞菌 生物膜 碳青霉烯类药物 转录组测序分析 

分 类 号：R978.1[医药卫生—药品]