College of Animal Science and Veterinary Medicine, Collaborative Innovation Center for Prevention and Control of Zoonoses, Jinzhou Medical University, Jinzhou, Liaoning, China.
Microbiol Spectr. 2024 Oct 3;12(10):e0034124. doi: 10.1128/spectrum.00341-24. Epub 2024 Sep 9.
The plasmid-mediated gene mcr-1 that makes bacteria resistant to the antibiotic colistin is spreading quickly, which means that colistin is no longer working well to treat Gram-negative bacterial infections. Herein, we utilized a computer-aided high-throughput screening drugs method to identify the natural product apigenin, a potential -protein inhibitor, which effectively enhanced the antimicrobial activity of colistin. Several assays, including a checkerboard minimum inhibitory concentration assay, a time-kill assay, the combined disk test, molecular simulation dynamics, and animal infection models assay, were conducted to verify whether apigenin enhanced the ability of colistin to fight Gram-negative bacterial infections. The results showed that apigenin improved the antimicrobial activity of colistin against multidrug-resistant infection. Moreover, apigenin not only did not increase the toxic effect of colistin but also had the ability to effectively inhibit the frequency of bacterial resistance mutations to colistin. Studies clearly elucidated that apigenin could interfere with the thermal stability of the protein by binding to the -1 protein. Additionally, the combination of apigenin and colistin could exert multiple effects, including disrupting bacterial membranes, the generation of bacterial nitric oxide and reactive oxygen species, as well as inhibiting bacterial adenosine triphosphate production. Furthermore, the addition of apigenin was able to significantly inhibit colistin-stimulated high expression levels of the bacterial mcr-1 gene. Finally, apigenin exhibited a characteristic anti-inflammatory effect while enhancing the antimicrobial activity of colistin against mcr-1-positive () infected animals. In conclusion, as a potential lead compound, apigenin is promising in combination with colistin in the future treatment of mcr-1-positive infections.IMPORTANCEThis study found that apigenin was able to inhibit the activity of the -1 protein using a high-throughput virtual screening method. Apigenin effectively enhanced the antimicrobial activity of colistin against multidrug-resistant , including mcr-1-positive strains, and . This study will provide new options and strategies for the future treatment of multidrug-resistant pathogen infections.
质粒介导的基因 mcr-1 使细菌对粘菌素这种抗生素产生耐药性，这意味着粘菌素治疗革兰氏阴性菌感染的效果已经大打折扣。在此，我们利用计算机辅助高通量筛选药物的方法，发现天然产物芹菜素是一种潜在的 -1 蛋白抑制剂，它可以有效增强粘菌素的抗菌活性。我们通过棋盘微量稀释法、时间杀伤试验、联合药敏纸片试验、分子模拟动力学和动物感染模型试验等方法，验证了芹菜素是否增强了粘菌素对抗革兰氏阴性菌感染的能力。结果表明，芹菜素提高了粘菌素对多重耐药感染的抗菌活性。此外，芹菜素不仅没有增加粘菌素的毒性作用，而且还具有有效抑制细菌对粘菌素耐药突变频率的能力。研究表明，芹菜素可以通过与 -1 蛋白结合来干扰蛋白的热稳定性。此外，芹菜素和粘菌素的联合使用可以发挥多种作用，包括破坏细菌膜、产生细菌一氧化氮和活性氧、抑制细菌三磷酸腺苷的产生。此外，添加芹菜素可以显著抑制粘菌素刺激的细菌 mcr-1 基因的高表达水平。最后，芹菜素在增强粘菌素对 mcr-1 阳性 （）感染动物的抗菌活性的同时，表现出一种抗炎作用。总之，作为一种潜在的先导化合物，芹菜素有望与粘菌素联合应用于未来 mcr-1 阳性感染的治疗。
本研究发现，芹菜素能够通过高通量虚拟筛选方法抑制 -1 蛋白的活性。芹菜素有效增强了粘菌素对多重耐药菌、包括 mcr-1 阳性株、和 的抗菌活性。本研究将为未来治疗多重耐药病原体感染提供新的选择和策略。
