抗菌肽BuforinⅡ衍生物对金黄色葡萄球菌细胞膜作用机制的研究

doi:10.11843/j.issn.0366-6964.2013.07.018

畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (7): 1124-1130.doi: 10.11843/j.issn.0366-6964.2013.07.018

抗菌肽BuforinⅡ衍生物对金黄色葡萄球菌细胞膜作用机制的研究

郝刚¹，乐国伟²，施用晖²

（1. 西南民族大学生命科学与技术学院，成都 610041；2. 江南大学食品学院，无锡 214122）

收稿日期:2012-11-29 出版日期:2013-07-23 发布日期:2013-07-23
作者简介:郝刚（1978-），男，重庆人，博士，主要从事抗菌肽的研究，Tel: 86-28-88434002, E-mail: indianahg@hotmail.com
基金资助:
国家自然科学基金（30871805；31172214）；中央高校基本科研业务基金（11NZYQN30）；十二五支撑计划（2012BAD33B05）

The Action Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡ on Staphylococcus aureus Membrane

HAO Gang¹, LE Guo-wei², SHI Yong-hui²

(1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China; 2. School of Food Science and Technology, Jiangnan University, Wuxi 214122, China)

Received:2012-11-29 Online:2013-07-23 Published:2013-07-23

摘要/Abstract

摘要：

为研究抗菌肽Buforin Ⅱ的衍生物BF2-A/B对金黄色葡萄球菌细胞膜的作用机制，用流式细胞仪分析BF2-A/B对金黄色葡萄球菌细胞膜通透性的影响，以及穿膜效率，用透射电镜观察细胞膜的完整性，激光共聚焦显微镜观察抗菌肽在胞质内的积累。结果显示，BF2-A/B都不显著引起PI流入细胞质内，但BF2-B处理的细胞PI着染阳性比例高于BF2-A。BF2-A处理20 min后的细胞膜依然完整，而BF2-B能使胞质内容物轻微泄漏。抗菌肽BF2-A/B都能显著穿透细胞膜，并且BF2-B的穿膜效率高于BF2-A。同时荧光共聚焦显微镜也证实抗菌肽在细胞质内的累积。研究结果说明BF2-A/B并不破坏金黄色葡萄球菌细胞膜，而是进入细胞内实施抗菌活性。BF2-B在穿透细胞膜的过程中对膜的扰动较大，导致了PI的少量流入以及细胞内容物的泄露，同时穿膜效率提高，抗菌活性更强。

Abstract:

In this paper, the action mechanism of BF2-A/B, two analogues of antimicrobial peptide BuforinⅡ, on Staphylococcus aureus membrane had been researched. The results of FACScan analysis implied that BF2-A/B did not induce the influx of PI into the S. aureus cells. The rate of positive cells stained by fluorescent probe after BF2-B treatment was slightly higher than that of BF2-A. These electron micrographs showed that after 20 min of treatment by BF2-A, S. aureus cells still retained the plasma membrane integrality; however, BF2-B could cause some slight leakages of cellular cytoplasmic contents. The results of FACS analysis displayed that both the peptides could penetrate the cells, and BF2-B penetrated the cells more efficiently. The visualization of confocal microscopy proved that FITC-labeled BF2-A and BF2-B penetrated the bacterial cell membrane and accumulated in the cytoplasm of the cell immediately. The results of research demonstrated that BF2-A/B didn’t destroy the cell membrane of G⁺bacteria, and then exert the antimicrobial activity after influx into cytoplasm. BF2-B slightly disturbed cell membrane causing influx of PI and leakage of cytoplasmic contents during peptide crossing phospholipids bilayer. Meanwhile, the cell-penetrating efficiency of BF2-B was accordingly enhanced, which caused that BF2-B displayed more excellent antimicrobial activity to S. aureus.

中图分类号:

郝刚，乐国伟，施用晖. 抗菌肽BuforinⅡ衍生物对金黄色葡萄球菌细胞膜作用机制的研究[J]. 畜牧兽医学报, 2013, 44(7): 1124-1130.

HAO Gang, LE Guo-wei, SHI Yong-hui. The Action Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡ on Staphylococcus aureus Membrane[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(7): 1124-1130.

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抗菌肽BuforinⅡ衍生物对金黄色葡萄球菌细胞膜作用机制的研究

The Action Mechanism of Two Analogues of the Antimicrobial Peptide BuforinⅡ on Staphylococcus aureus Membrane

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