摘要

       抗生素耐药性的出现和日益流行对人类健康构成了全球公共风险，其中质粒介导的水平基因转移（HGT）在这一过程中发挥着重要作用。然而，非抗生素药物与抗生素抗性基因（ARG）在环境中的传播之间的关系仍知之甚少。在此，我们旨在研究对乙酰氨基酚（世界上最常见的解热镇痛药之一）在质粒携带的 ARG 共轭转移中的作用。使用携带 RP4-7 质粒的工程细菌大肠杆菌 DH5α 作为供体细菌和大肠杆菌 EC600 作为受体细菌，我们发现低浓度（包括环境相关浓度）的对乙酰氨基酚显着促进了质粒介导的结合转移。在携带 mcr-1 或 tet(X4) 的各种临床质粒中也观察到类似的促进作用，这两种关键耐药决定因素分别赋予细菌对粘菌素和替加环素的耐药性。进一步的机械实验表明，补充对乙酰氨基酚可增加细胞膜通透性，刺激 ROS 产生，诱导 SOS 反应并加强结合桥，从而共同促进结合转移。此后，通过 RT-PCR 验证，与上述方式相关的基因的表达水平在暴露于对乙酰氨基酚后显着上调。总之，我们的研究表明对乙酰氨基酚可以大大加速细菌中的结合转移。这些发现为 ARG 的传播提供了新的见解，并暗示了在临床环境中使用对乙酰氨基酚引起的潜在风险。

       The emergence and increasing prevalence of antibiotic resistance pose a global public risk for human health, among which plasmid-mediated horizontal gene transfer (HGT) play an important role in this process. However, the relationship between non-antibiotic drugs and dissemination of antibiotic resistance genes (ARGs) in the environment is still poorly understood. Herein, we aimed to investigate the roles of acetaminophen, one of the most common antipyretic analgesics worldwide, in the conjugative transfer of plasmid-borne ARGs. Using the engineering bacteria Escherichia coli DH5α carrying the RP4-7 plasmid as donor bacteria and E. coli EC600 as recipient bacteria, we found that acetaminophen at low concentrations including environmentally relevant concentrations significantly promoted the plasmid-mediated conjugative transfer. Similar promotion effects were also observed in various clinical plasmids that carrying mcr-1 or tet(X4), two critical resistance determinants that confer bacterial resistance to colistin and tigecycline, respectively. Further mechanistic experiments suggested that acetaminophen supplementation increased cell membrane permeability, stimulated ROS production, induced SOS response and strengthened conjugation bridge, thereby collectively contributing to the enhanced conjugative transfer. Thereafter, validated by RT-PCR, the expression levels of genes related to these manners above were conspicuously upregulated after exposure to acetaminophen. Taken together, our study demonstrated that acetaminophen could drastically accelerate the conjugative transfer in bacteria. These findings provide new insights into the spread of ARGs and imply a potential risk elicited by the use of acetaminophen in the clinical setting.

https://www.sciencedirect.com/science/article/abs/pii/S0048969721019860