Key Laboratory of Major Diseases in Children, Ministry of Education, National Clinical Research Center for Respiratory Diseases, National Key Discipline of Pediatrics, Laboratory of Infection and Microbiology, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing 100045, China.
Beijing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Microbiol Res. 2024 Nov;288:127872. doi: 10.1016/j.micres.2024.127872. Epub 2024 Aug 11.
Antimicrobial resistance has been an increasingly serious threat to global public health. The contribution of non-antibiotic pharmaceuticals to the development of antibiotic resistance has been overlooked. Our study found that the anti-inflammatory drug phenylbutazone could protect P. aeruginosa against antibiotic mediated killing by binding to the efflux pump regulator MexR. In this study, antibiotic activity against P. aeruginosa alone or in combination with phenylbutazone was evaluated in vitro and in vivo. Resazurin accumulation assay, transcriptomic sequencing, and PISA assay were conducted to explore the underlying mechanism for the reduced antibiotic susceptibility caused by phenylbutazone. Then EMSA, ITC, molecular dynamic simulations, and amino acid substitutions were used to investigate the interactions between phenylbutazone and MexR. We found that phenylbutazone could reduce the susceptibility of P. aeruginosa to multiple antibiotics, including parts of β-lactams, fluoroquinolones, tetracyclines, and macrolides. Phenylbutazone could directly bind to MexR, then promote MexR dissociating from the mexA-mexR intergenic region and de-repress the expression of MexAB-OprM efflux pump. The overexpressed MexAB-OprM pump resulted in the reduced antibiotic susceptibility. And the His41 and Arg21 residues of MexR were involved in the phenylbutazone-MexR interaction. We hope this study would imply the potential risk of antibiotic resistance caused by non-antibiotic pharmaceuticals.
抗微生物药物耐药性一直是对全球公共卫生的一个日益严重的威胁。非抗生素药物对抗微生物药物耐药性发展的贡献一直被忽视。我们的研究发现，抗炎药苯丁唑酮可以通过与外排泵调节剂 MexR 结合来保护铜绿假单胞菌免受抗生素介导的杀伤。在这项研究中，我们评估了单独使用抗生素或与苯丁唑酮联合使用对铜绿假单胞菌的体外和体内活性。进行了 Resazurin 积累测定、转录组测序和 PISA 测定，以探索苯丁唑酮导致抗生素敏感性降低的潜在机制。然后，进行了 EMSA、ITC、分子动力学模拟和氨基酸取代，以研究苯丁唑酮与 MexR 之间的相互作用。我们发现，苯丁唑酮可以降低铜绿假单胞菌对多种抗生素的敏感性，包括部分β-内酰胺类、氟喹诺酮类、四环素类和大环内酯类抗生素。苯丁唑酮可以直接与 MexR 结合，然后促进 MexR 从 mexA-mexR 基因间区解离，解除 MexAB-OprM 外排泵的抑制表达。过度表达的 MexAB-OprM 泵导致抗生素敏感性降低。MexR 的 His41 和 Arg21 残基参与了苯丁唑酮-MexR 相互作用。我们希望本研究能暗示非抗生素药物引起抗微生物药物耐药性的潜在风险。
