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宿主防御肽模拟聚合物与蛋白酶K联用抗生物被膜

毕玉芳 武月铭 刘士琦 钱宇芯 刘润辉

毕玉芳, 武月铭, 刘士琦, 钱宇芯, 刘润辉. 宿主防御肽模拟聚合物与蛋白酶K联用抗生物被膜[J]. 功能高分子学报. doi: 10.14133/j.cnki.1008-9357.20210523001
引用本文: 毕玉芳, 武月铭, 刘士琦, 钱宇芯, 刘润辉. 宿主防御肽模拟聚合物与蛋白酶K联用抗生物被膜[J]. 功能高分子学报. doi: 10.14133/j.cnki.1008-9357.20210523001
BI Yufang, WU Yueming, LIU Shiqi, QIAN Yuxin, LIU Runhui. Combination of Host Defense Peptide Mimicking Peptide Polymer and Proteinase K against Biofilms[J]. Journal of Functional Polymers. doi: 10.14133/j.cnki.1008-9357.20210523001
Citation: BI Yufang, WU Yueming, LIU Shiqi, QIAN Yuxin, LIU Runhui. Combination of Host Defense Peptide Mimicking Peptide Polymer and Proteinase K against Biofilms[J]. Journal of Functional Polymers. doi: 10.14133/j.cnki.1008-9357.20210523001

宿主防御肽模拟聚合物与蛋白酶K联用抗生物被膜

doi: 10.14133/j.cnki.1008-9357.20210523001
基金项目: 国家自然科学基金(22075078, 21861162010);上海市自然科学基金(18ZR1410300);上海市优秀青年学术带头人(20XD1421400)
详细信息
    作者简介:

    毕玉芳(1996—),女,硕士生,研究方向为生物医用高分子材料。E. mail:bi1521673@163.com

    通讯作者:

    刘润辉,E-mail:rliu@ecust.edu.cn

  • 中图分类号: R318.08

Combination of Host Defense Peptide Mimicking Peptide Polymer and Proteinase K against Biofilms

  • 摘要: 采用双(三甲基硅基)胺基锂(LiHMDS)引发的α-氨基酸N-羧酸酐(NCA)快速开环聚合方法合成宿主防御肽模拟聚合物,该聚合物呈现窄分子量分布。抗菌实验结果表明, 该聚合物对耐甲氧西林金黄色葡萄球菌(MRSA)浮游菌和持留菌有高杀菌活性,且遵循破膜杀菌机制。实验证明,1.25 U/mL的蛋白酶K在人工尿液环境中可分散MRSA成熟生物被膜。蛋白酶K与聚合物协同作用实现了人工尿液中成熟生物被膜的清除。

     

  • 图  1  蛋白酶K和P(DLL-co-BLG)联合作用清除MRSA生物被膜和持留菌的示意图

    Figure  1.  Schematic diagram of combined action of protease K and P(DLL-co-BLG) to eradicate MRSA biofilms and persister cells

    图  2  多肽聚合物P(DLL-co-BLG)的合成

    Figure  2.  Synthesis of the peptide polymer P(DLL-co-BLG)

    图  3  P(DLL-co-BLG)的GPC曲线(a)和1H-NMR图谱(b)

    Figure  3.  GPC curve (a) and 1H-NMR spectrum (b) of P(DLL-co-BLG)

    图  4  P(DLL-co-BLG)对金黄色葡萄球菌的MIC值(a)和对S. aureus USA300的MBC值(b)

    Figure  4.  MIC against S. aureus (a) and MBC against S. aureus USA300 (b) of P(DLL-co-BLG)

    图  5  P(DLL-co-BLG)与S. aureus USA300作用后DiSC3(5)荧光强度随时间的变化

    Figure  5.  Fluorescence of DiSC3(5) in the presence of S. aureus USA300 and P(DLL-co-BLG) as a function of the incubation time

    图  6  浓度为4×MIC的P(DLL-co-BLG)对S. aureus USA300持留菌的杀菌动力学曲线

    Figure  6.  Time-kill curves for P(DLL-co-BLG) with final concentrations of 4×MIC against S. aureus USA300 persister cells

    图  7  (a)不同浓度的蛋白酶K对人工尿中S. aureus USA300成熟生物膜细菌活力的影响;(b)MTT法获得人工尿中P(DLL-co-BLG)对S. aureus USA300成熟生物被膜的清除活性(*P<0.05, **P<0.01, ***P<0.001,显著性差异是通过与未经聚合物处理的对照组对比得到的);(c)人工尿液介质中,蛋白酶K和P(DLL-co-BLG)联合作用后,S. aureus USA300成熟生物被膜内部的细菌活力(图中小写字母表示各实验组间有显著性差异(P<0.05,柱状图上方为药物处理和未处理的经过MTT染色后的生物被膜的图像)

    Figure  7.  (a) Effects of proteinase K on mature S. aureus USA300 biofilms in artificial urine; (b) Eradication activity of P(DLL-co-BLG) in artificial urine against mature S. aureus USA300 biofilms obtained by MTT assay (*P<0.05, **P<0.01, ***P<0.001, significant difference analysis results were obtained when compared with the control groups that had not been treated with polymers);(c) Remaining cell viability within mature S. aureus USA300 biofilm after the combined action of protease K and P(DLL-co-BLG) in artificial urine (Different lowercase letters indicate significant difference (P< 0.05) between different experimental groups, above the bar diagram are the corresponding images of treated and untreated biofilms after MTT staining and precipitation dissolution)

    图  8  蛋白酶K和P(DLL-co-BLG)联合作用后,S. aureus USA300成熟生物被膜的活/死染色图像

    Figure  8.  Live-dead staining fluorescence micrographs of mature S. aureus USA300 biofilms treated with the combination of protease K and P(DLL-co-BLG)

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出版历程
  • 收稿日期:  2021-05-23
  • 网络出版日期:  2021-07-19

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