康替唑胺、利奈唑胺对革兰阳性菌的体外抗菌活性及生物被膜抑制机制  

Inhibitory effects of contezolid and linezolid on growth of Gram-positive bacteria and biofilm formation

作　　者：曹心怡 沈宗霖 李桂秋 林志伟 郑金鑫 余治健 魏影 CAO Xinyi;SHEN Zonglin;LI Guiqiu;LIN Zhiwei;ZHENG Jinxin;YU Zhijian;WEI Ying(Clinical Medical College,Jiamusi University,Jiamusi 154003,China;不详)

机构地区：[1]佳木斯大学临床医学院,黑龙江佳木斯154003 [2]华中科技大学协和深圳医院深圳市内源性感染重点实验室 [3]华中科技大学协和深圳医院检验医学中心 [4]黑龙江省卫生健康管理服务评价中心

出　　处：《山东医药》2023年第29期36-40,共5页Shandong Medical Journal

基　　金：国家自然科学基金项目(81902033);广东省自然科学基金项目(2020A1515011049);深圳市教育科学研究课题项目(NS2022046)。

摘　　要：目的探讨康替唑胺、利奈唑胺对常见革兰阳性菌金黄色葡萄球菌、粪肠球菌的体外抗菌活性,初步分析其抑制生物被膜的相关机制。方法取革兰阳性菌金黄色葡萄球菌、粪肠球菌,采用微量肉汤稀释法测定康替唑胺、利奈唑胺对金黄色葡萄球菌、粪肠球菌的最小抑菌浓度(MIC)。通过生长曲线检测不同药物浓度下细菌生长情况,在不抑制细菌生长的药物浓度下通过结晶紫染色观察细菌生物被膜形成情况及生物被膜内细菌存活率。采用蛋白质组学筛选康替唑胺、利奈唑胺作用下革兰阳性菌生物被膜差异表达蛋白,基因本体(GO)功能注释及京都基因与基因组百科全书(KEGG)通路分析差异表达蛋白功能,蛋白质相互作用网络(PPI)观察蛋白之间相互作用关系。结果康替唑胺及利奈唑胺对金黄色葡萄球菌、粪肠球菌表现出广谱抗菌活性,被选入实验所用菌株MIC均≤4µg/mL,生长曲线结果显示康替唑胺、利奈唑胺在1/2×MIC并未影响细菌生长,结晶紫染色及生物被膜存活菌计数显示亚抑菌浓度下两种药物能够抑制金黄色葡萄球菌、粪肠球菌生物被膜形成;康替唑胺、利奈唑胺作用下革兰阳性菌生物被膜差异表达蛋白分别为290、222个,差异蛋白主要涉及rpmJ、rplE、SasG、luxS、ald1、D-丙氨酸氨基转移酶等。GO注释分析显示,康替唑胺作用下差异表达蛋白主要与丙酮酸代谢过程、氨基酸代谢等过程相关;利奈唑胺作用下差异表达蛋白主要涉及嘧啶代谢过程、嘌呤代谢、氨基酸代谢等过程。KEGG分析显示,康替唑胺作用下差异表达蛋白主要涉及萘降解、嘧啶代谢、糖酵解、核糖体切除修复等过程;利奈唑胺作用下差异表达蛋白主要涉及氨基酸降解、脂肪酸降解、嘌呤代谢、戊糖和葡萄糖磷酸盐转化过程。PPI网络分析显示,两种抗生素作用下的差异表达蛋白主要与核糖体相关。结论康�Objective To investigate the antibacterial activity of contezolid and linezolid against common Gram-posi⁃tive bacteria and to preliminarily analyze the mechanism of their inhibition of biofilm.Methods Staphylococcus aureus(S.aureus)and Enterococcus faecalis(E.faecalis)were taken as the research objects.We used the broth dilution meth⁃od to determine the minimum inhibitory concentration(MIC)of contezolid and linezolid against S.aureus and E.faecalis.The growth curve was used to detect the growth of bacteria under different drug concentrations,and the formation of bacteri⁃al biofilm and the survival rate of bacteria in the biofilm were observed by crystal violet staining under the drug concentra⁃tion that did not inhibit the growth of bacteria.Proteomics was used to screen the differentially expressed proteins in the biofilm of Gram-positive bacteria under the action of contezolid and linezolid.Functional annotation of gene ontology(GO),Kyoto Encylopaedia of Genes and Genomes(KEGG),and protein interaction network(PPI)was used to observe the interaction between proteins.Results Contezolid and linezolid showed broad-spectrum antibacterial activity against S.aureus and E.faecalis,and their MICs were both less than 4μg/mL.The growth curve showed that contezolid and line⁃zolid at 1/2×MIC had no effect on bacterial growth.The result of crystal violet staining and the count of viable bacteria in biofilm showed that both drugs could inhibit the formation of S.aureus and E.faecalis biofilm at sub-inhibitory concentra⁃tion.There were 290 and 222 differentially expressed proteins in the biofilm of Gram-positive bacteria under the action of contezolid and linezolid.The differential proteins mainly involved Rpm,rplE,SasG,Luxs,ald 1,D-alanine aminotrans⁃ferase,etc.GO analysis showed that the differentially expressed proteins were mainly related to pyruvate metabolism and amino acid metabolism under the action of contezolid.The differential expression of proteins under linezolid mainly in⁃volved pyrimidine metab

关 键 词：恶唑烷酮 康替唑胺 利奈唑胺 革兰阳性菌 金黄色葡萄球菌 粪肠球菌 抗菌活性 生物被膜 

分 类 号：R915[医药卫生—微生物与生化药学]