Canè Carolina, Gallucci Noemi, Amoresano Angela, Fontanarosa Carolina, Paduano Luigi, De Gregorio Eliana, Duilio Angela, Di Somma Angela
Department of Chemical Sciences, University of Naples "Federico II" Via Cinthia 4, 80126, Napoli, Italy.
National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d'Oro 305, 00136, Rome, Italy.
Biochem Biophys Rep. 2024 Aug 15;39:101808. doi: 10.1016/j.bbrep.2024.101808. eCollection 2024 Sep.
The evolution of methicillin-resistant () has required the development of new antimicrobial agents and new approaches to prevent and overcome drug resistance. AntiMicrobial Peptides (AMPs) represent promising alternatives due to their rapid bactericidal activity and their broad-spectrum of action against a wide range of microorganisms. The amphibian Temporins constitute a well-known family of AMPs with high antibacterial properties against both Gram-positive and Gram-negative bacteria. In this paper, we evaluated the in vivo effect of Temp-L on performing morphological studies using Transmission Electron Microscopy (TEM) that revealed the occurrence of protrusions from the cell surface. The formation of vesicle-like structure was confirmed by Dynamic Light Scattering (DLS). The global effect of Temp-L on () was deeply investigated by differential proteomics leading to the identification of up-regulated proteins involved in the synthesis of the cell membrane and fatty acids, and down-regulated virulence factors. GC-MS analysis suggested a possible protective response mechanism implemented by the bacterium after treatment with Temp-L, as the synthesis of fatty acids was increased. Adhesion and invasion assays on eukaryotic cells confirmed a reduced virulence of following treatment with Temp-L. These results suggested the targeting of virulence factors as novel strategy to replace traditional antimicrobial agents that can be used to treat infections, especially infections caused by the resistant pathogen .
耐甲氧西林金黄色葡萄球菌（MRSA）的演变促使了新型抗菌药物的研发以及预防和克服耐药性的新方法的出现。抗菌肽（AMPs）因其快速杀菌活性和对多种微生物的广谱抗菌作用而成为有前景的替代物。两栖动物的天蚕素构成了一类著名的抗菌肽家族，对革兰氏阳性菌和革兰氏阴性菌均具有高抗菌特性。在本文中，我们使用透射电子显微镜（TEM）进行形态学研究，评估了Temp-L对MRSA的体内作用，结果显示细胞表面出现了突起。动态光散射（DLS）证实了囊泡样结构的形成。通过差异蛋白质组学深入研究了Temp-L对MRSA的整体作用，从而鉴定出参与细胞膜和脂肪酸合成的上调蛋白以及下调的毒力因子。气相色谱-质谱联用（GC-MS）分析表明，用Temp-L处理后，细菌可能实施了一种保护性反应机制，因为脂肪酸的合成增加了。对真核细胞的黏附和侵袭试验证实，用Temp-L处理后MRSA的毒力降低。这些结果表明，将毒力因子作为一种新策略来替代传统抗菌药物，可用于治疗感染，尤其是由耐药病原体MRSA引起的感染。
