Wang Qing, Xu Yanzhao, Hu Jianhe
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology Xinxiang 453003 China
RSC Adv. 2022 May 13;12(23):14485-14491. doi: 10.1039/d2ra01363k. eCollection 2022 May 12.
To explore the potential intracellular mechanism of the antimicrobial peptide HJH-3 in killing , a DNA blocking test and scanning electron microscopy (SEM) were used to determine the ability of the peptide to bind bacterial DNA . Laser confocal analysis and electron microscopy were used to observe the binding of antimicrobial peptide HJH-3 and DNA, and flow cytometry was used to analyze the effect of antimicrobial peptides on cell division . The results showed that HJH-3 can bind to DNA to block the diffusion and migration of DNA in agarose gel. Laser confocal microscopy revealed that antimicrobial peptide HJH-3 penetrated the bacterial cell membrane and bound with bacterial DNA. Transmission electron microscopy showed that antimicrobial peptide HJH-3 aggregated in the nucleoid of cells, and through a channel in the membrane destroyed by the antimicrobial peptide, DNA and other intracellular contents were excreted, and polymerized DNA was fragmented. The results of the flow cytometry analysis confirmed that the death rate of increased significantly after exposure to antimicrobial peptide HJH-3 and increased with increasing antimicrobial peptide concentration. These results suggest that AMP HJH-3 may be a candidate antimicrobial agent to treat infectious diseases caused by .
为探究抗菌肽HJH-3杀菌的潜在细胞内机制，采用DNA阻断试验和扫描电子显微镜（SEM）来确定该肽结合细菌DNA的能力。利用激光共聚焦分析和电子显微镜观察抗菌肽HJH-3与DNA的结合情况，并运用流式细胞术分析抗菌肽对细胞分裂的影响。结果表明，HJH-3能与DNA结合，从而阻断DNA在琼脂糖凝胶中的扩散和迁移。激光共聚焦显微镜显示，抗菌肽HJH-3穿透细菌细胞膜并与细菌DNA结合。透射电子显微镜表明，抗菌肽HJH-3聚集在细胞的类核中，通过抗菌肽破坏的细胞膜通道，DNA和其他细胞内物质被排出，聚合的DNA发生片段化。流式细胞术分析结果证实，暴露于抗菌肽HJH-3后，死亡率显著增加，且随抗菌肽浓度增加而升高。这些结果表明，AMP HJH-3可能是治疗由[病原体名称缺失]引起的传染病的候选抗菌剂。
