Combined Systems Approaches Reveal a Multistage Mode of Action of a Marine Antimicrobial Peptide against Pathogenic Escherichia coli and Its Protective Effect against Bacterial Peritonitis and Endotoxemia

文献类型： 外文期刊

第一作者： Wang, Xiumin

作者： Wang, Xiumin;Teng, Da;Mao, Ruoyu;Yang, Na;Hao, Ya;Wang, Jianhua;Wang, Xiumin;Teng, Da;Mao, Ruoyu;Yang, Na;Hao, Ya;Wang, Jianhua

作者机构：

关键词： marine antimicrobial peptides;N4;mechanism;Escherichia coli;lipopolysaccharide

期刊名称：ANTIMICROBIAL AGENTS AND CHEMOTHERAPY （ 影响因子：5.191； 五年影响因子：5.346 ）

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收录情况： SCI

摘要： A marine arenicin-3 derivative, N4, displayed potent antibacterial activity against Gram-negative bacteria, but its antibacterial mode of action remains elusive. The mechanism of action of N4 against pathogenic Escherichia coli was first researched by combined cytological and transcriptomic techniques in this study. The N4 peptide permeabilized the outer membrane within 1 min, disrupted the plasma membrane after 0.5 h, and localized in the cytoplasm within 5 min. Gel retardation and circular dichroism (CD) spectrum analyses demonstrated that N4 bound specifically to DNA and disrupted the DNA conformation from the B type to the C type. N4 inhibited 21.1% of the DNA and 20.6% of the RNA synthesis within 15 min. Several hallmarks of apoptosis-like cell death were exhibited by N4-induced E. coli, such as cell cycle arrest in the replication (R) and division(D) phases, reactive oxygen species production, depolarization of the plasma membrane potential, and chromatin condensation within 0.5 h. Deformed cell morphology, disappearance of the plasma membrane, leakage of the contents, and ghost cell formation were demonstrated by transmission electron microscopy, and nearly 100% of the bacteria were killed by N4. A total of 428 to 663 differentially expressed genes are involved in the response to N4, which are associated mainly with membrane biogenesis (53.9% to 56.7%) and DNA binding (13.3% to 14.9%). N4-protected mice that were lethally challenged with lipopolysaccharide (LPS) exhibited reduced levels of interleukin-6 (IL-6), IL-1 beta, and tumor necrosis factor alpha (TNF-alpha) in serum and protected the lungs from LPS-induced injury. These data facilitate an enhanced understanding of the mechanisms of marine antimicrobial peptides (AMPs) against Gram-negative bacteria and provide guidelines in developing and applying novel multitarget AMPs in the field of unlimited marine resources as therapeutics.

分类号： R45

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