多黏菌素B联合新型β-内酰胺酶抑制剂复方制剂对耐多黏菌素B细菌生物膜的影响  

Effect of polymyxin B combined with novelβ-lactam/β-lactamase inhibitor combinations on biofilms of polymyxin B-resistant bacteria

作　　者：田淼梅 郭思维 颜冰倩 蒋蓉 徐兵 李昕 TIAN Miaomei;GUO Siwei;YAN Bingqian;JIANG Rong;XU Bing;LI Xin(Department of Pharmacy,The Third Hospital of Changsha,Changsha 410015,China;Changsha Institute of Clinical Application of Antimicrobial agents,Changsha 410015,China;Hunan Provincial Key Laboratory of Anti-Resistance Microbial Drugs,Changsha 410015,China)

机构地区：[1]长沙市第三医院药学部,湖南长沙410015 [2]长沙抗菌药物临床研究所,湖南长沙410015 [3]抗耐药微生物药物湖南省重点实验室,湖南长沙410015

出　　处：《中国感染控制杂志》2025年第1期58-66,共9页Chinese Journal of Infection Control

基　　金：抗耐药微生物药物湖南省重点实验室基金项目(2023TP1013)。

摘　　要：目的探讨多黏菌素B联合新型β-内酰胺酶抑制剂复方制剂对耐多黏菌素B铜绿假单胞菌和肺炎克雷伯菌生物膜的活性及机制。方法采用微量肉汤稀释法和卡尔加里生物膜装置测定所有抗菌药物的最低抑菌浓度(MIC)、最低生物膜抑制浓度(MBIC)和最低生物膜根除浓度(MBEC);采用结晶紫染色法评估亚MIC剂量下多黏菌素B联合头孢吡肟/阿维巴坦、头孢他啶/阿维巴坦、美罗培南/阿维巴坦、氨曲南/阿维巴坦、美罗培南/法硼巴坦及亚胺培南/雷利巴坦抑制生物膜形成和根除成熟生物膜的效果;筛选出抗生物膜活性最佳的联合方案,采用硫酸苯酚法、细菌运动试验以及群体感应抑制试验初步探讨该联合方案的抗生物膜机制。结果所有抗菌药物的MBIC和MBEC均高于MIC,基于多黏菌素B的联合方案均能抑制铜绿假单胞菌和肺炎克雷伯菌生物膜形成并根除成熟的生物膜,其中多黏菌素B联合头孢吡肟/阿维巴坦抑制率和根除率最高,分别为67.99%~90.16%、58.26%~63.86%。多黏菌素B联合头孢吡肟/阿维巴坦可抑制肺炎克雷伯菌的胞外多糖,抑制率为34.04%~61.10%,该组合还能减小细菌的泳动和蹭行运动直径。头孢吡肟/阿维巴坦单药对群体感应信号分子的抑制作用呈浓度依赖性,与多黏菌素B联合后,抑制作用与单药效果一致。结论多黏菌素B和头孢吡肟/阿维巴坦可能是临床治疗多黏菌素B耐药菌株生物膜相关严重感染的潜在方案,其作用机制可能与抑制细菌胞外多糖和运动能力有关。Objective To explore the activity and mechanism of polymyxin B combined with novelβ-lactam/β-lactamase inhibitor combinations on biofilm of polymyxin B-resistant Pseudomonas aeruginosa and Klebsiella pneumo-niae.Methods The minimum inhibitory concentration(MIC),minimum biofilm inhibitory concentration(MBIC),and minimum biofilm eradication concentration(MBEC)of all antimicrobial agents were determined by micro-broth dilution method and MBEC TM assay.The crystal violet staining method was adopted to evaluate the effect of polymyxin B combined with cefepime/avibactam,ceftazidime/avibactam,meropenem/avibactam,aztreonam/avibactam,meropenem/vaborbactam,and imipenem/relebactam at sub-MIC doses on inhibition of biofilm formation and eradication of mature biofilm.The best combination scheme for anti-biofilm activity was screened out,and anti-biofilm mechanism of this combination scheme was preliminarily explored with phenol-sulfuric acid method,bacterial motility test,and quorum sensing inhibition test.Results The MBIC and MBEC of all antimicrobial agents were higher than MIC.The combination regimen based on polymyxin B could inhibit the formation of biofilms and eradicate mature biofilm in Pseudomonas aeruginosa and Klebsiella pneumoniae.The combination of polymyxin B with cefepime/avibactam had the highest inhibition and eradication rates,ranging 67.99%-90.16%and 58.26%-63.86%,respectively.The combination of polymyxin B and cefepime/avibactam could inhibit the extracellular polysaccharides of Klebsiella pneumoniae,with inhibition rates of 34.04%-61.10%,this combination could also reduce the swimming and twitching motility diameters of bacteria.Cefepime/avibactam monotherapy on quorum sen-sing signaling molecules presented concentration dependent inhibitory effect,and when combined with polymyxin B,the inhibitory effect was consistent with that of monotherapy.Conclusion Polymyxin B and cefepime/avibactam may be potential scheme for clinical treatment for severe biofilm-associated infection caused by polymyxin B-resista

关 键 词：生物膜 铜绿假单胞菌 肺炎克雷伯菌 耐多黏菌素B 新型β-内酰胺酶抑制剂复方制剂 

分 类 号：R181.32[医药卫生—流行病学] R378[医药卫生—公共卫生与预防医学]