4-(金刚烷基-1)-1-(1-氨基丁基)苯对耐甲氧西林金黄色葡萄球菌的抗菌活性研究

Q4 Biochemistry, Genetics and Molecular Biology
N. Hrynchuk, L. Zelena, T. Bukhtiarova, N. Vrynchanu, L. Ishchenko, E. Vazhnichaya
{"title":"4-(金刚烷基-1)-1-(1-氨基丁基)苯对耐甲氧西林金黄色葡萄球菌的抗菌活性研究","authors":"N. Hrynchuk, L. Zelena, T. Bukhtiarova, N. Vrynchanu, L. Ishchenko, E. Vazhnichaya","doi":"10.15407/microbiolj84.03.039","DOIUrl":null,"url":null,"abstract":"Staphylococcus aureus is a widespread opportunistic pathogen, causing community-acquired and nosocomial infections with both acute and chronic recurrent courses. The process of chronicity of the disease is provided by biofilms. Features of the structure and functioning of biofilms, in particular the presence of matrix, quorum sensing systems, persistent cells, and efflux pumps, provide microbial communities with resistance to antimicrobial drugs under their action in therapeutic concentrations. The insufficient eff ectiveness of modern antimicrobial chemotherapy against biofi lm microorganisms indicates the urgency of the problem to search for compounds with antibiofilm activity that can affect various stages of the biofilm formation and the formed biofilm. The aim of the study is to establish the antibiofilm activity of 4-(adamantyl-1)-1-(1-aminobutyl) benzol against methicillin-resistant S. aureus (MRSA) and to determine the mechanism of its action. Methods. The ability of adamantane-containing compound 4-(adamantyl-1)-1-(1-aminobutyl) benzol (AM-166) to prevent biofilm formation and destroy the formed biofilm of S. aureus was investigated on polystyrene plates by the sorption of gentian violet on its structures followed with desorption of the dye into the organic solvent. The viability of S. aureus cells at the first stage of biofilm formation and in the composition of mature biofilms was evaluated using specific dyes for living (acridine orange) and non-viable (propidium iodide) cells. Detection of genes responsible for antibiotic resistance and biofi lm formation was performed by the polymerase chain reaction (PCR) with detection of PCR products in agarose gel. Evaluation of the effect of AM-166 on the expression of genes regulating the biofilm formation (ica, agrA, sarA, and sigB) was investigated by the real-time PCR and semi-quantitative PCR. Results. It was found that the compound AM-166 shows activity against S. aureus biofilm formation. The most pronounced effect was registered at a concentration of 5.0 minimum inhibitory concentration (MIC) (92.3%.) Under the action of AM-166 on the formed 2-day biofilms, their destruction was marked: the biomass decreases by 30.9% at 5.0 MIC. According to the results of fluorescence microscopy, the adamantane derivative at 5.0 MIC helps to reduce the number of viable cells at different stages of formation of the S. aureus biofilm. The results of molecular genetic studies indicate that the ica gene expression is significantly inhibited by the action of subinhibitory concentrations of the compound AM-116. No significant changes in the expression of sarA, agrA, and sigB genes were registered. Conclusions. Experiments on the effect of adamantane derivative on S. aureus biofilms showed that the most pronounced activity of AM-116 was observed at the stage of biofilm formation, as evidenced by the inhibition of transcriptional activity of the ica gene responsible for early stages of the biofilm formation, in particular the adhesion of planktonic cells to the substrate.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"1 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Antibiofilm Activity of 4-(Adamantyl-1)-1-(1-Aminobutyl) Benzol against Methicillin-Resistant Staphylococcus aureus\",\"authors\":\"N. Hrynchuk, L. Zelena, T. Bukhtiarova, N. Vrynchanu, L. Ishchenko, E. Vazhnichaya\",\"doi\":\"10.15407/microbiolj84.03.039\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Staphylococcus aureus is a widespread opportunistic pathogen, causing community-acquired and nosocomial infections with both acute and chronic recurrent courses. The process of chronicity of the disease is provided by biofilms. Features of the structure and functioning of biofilms, in particular the presence of matrix, quorum sensing systems, persistent cells, and efflux pumps, provide microbial communities with resistance to antimicrobial drugs under their action in therapeutic concentrations. The insufficient eff ectiveness of modern antimicrobial chemotherapy against biofi lm microorganisms indicates the urgency of the problem to search for compounds with antibiofilm activity that can affect various stages of the biofilm formation and the formed biofilm. The aim of the study is to establish the antibiofilm activity of 4-(adamantyl-1)-1-(1-aminobutyl) benzol against methicillin-resistant S. aureus (MRSA) and to determine the mechanism of its action. Methods. The ability of adamantane-containing compound 4-(adamantyl-1)-1-(1-aminobutyl) benzol (AM-166) to prevent biofilm formation and destroy the formed biofilm of S. aureus was investigated on polystyrene plates by the sorption of gentian violet on its structures followed with desorption of the dye into the organic solvent. The viability of S. aureus cells at the first stage of biofilm formation and in the composition of mature biofilms was evaluated using specific dyes for living (acridine orange) and non-viable (propidium iodide) cells. Detection of genes responsible for antibiotic resistance and biofi lm formation was performed by the polymerase chain reaction (PCR) with detection of PCR products in agarose gel. Evaluation of the effect of AM-166 on the expression of genes regulating the biofilm formation (ica, agrA, sarA, and sigB) was investigated by the real-time PCR and semi-quantitative PCR. Results. It was found that the compound AM-166 shows activity against S. aureus biofilm formation. The most pronounced effect was registered at a concentration of 5.0 minimum inhibitory concentration (MIC) (92.3%.) Under the action of AM-166 on the formed 2-day biofilms, their destruction was marked: the biomass decreases by 30.9% at 5.0 MIC. According to the results of fluorescence microscopy, the adamantane derivative at 5.0 MIC helps to reduce the number of viable cells at different stages of formation of the S. aureus biofilm. The results of molecular genetic studies indicate that the ica gene expression is significantly inhibited by the action of subinhibitory concentrations of the compound AM-116. No significant changes in the expression of sarA, agrA, and sigB genes were registered. Conclusions. Experiments on the effect of adamantane derivative on S. aureus biofilms showed that the most pronounced activity of AM-116 was observed at the stage of biofilm formation, as evidenced by the inhibition of transcriptional activity of the ica gene responsible for early stages of the biofilm formation, in particular the adhesion of planktonic cells to the substrate.\",\"PeriodicalId\":18628,\"journal\":{\"name\":\"Mikrobiolohichnyi zhurnal\",\"volume\":\"1 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-12-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Mikrobiolohichnyi zhurnal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.15407/microbiolj84.03.039\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mikrobiolohichnyi zhurnal","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.15407/microbiolj84.03.039","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0

摘要

金黄色葡萄球菌是一种广泛存在的机会性病原体,可引起急性和慢性复发的社区获得性和医院感染。疾病的慢性过程是由生物膜提供的。生物膜的结构和功能特征,特别是基质、群体感应系统、持久细胞和外排泵的存在,使微生物群落在治疗浓度下对抗菌药物产生耐药性。现代抗微生物化疗对生物膜微生物的有效性不足,表明寻找具有抗生物膜活性的化合物能够影响生物膜形成的各个阶段和形成的生物膜的问题的紧迫性。本研究的目的是建立4-(金刚烷基-1)-1-(1-氨基丁基)苯对耐甲氧西林金黄色葡萄球菌(MRSA)的抗菌膜活性,并确定其作用机制。方法。在聚苯乙烯板上研究了含金刚烷的化合物4-(金刚烷基-1)-1-(1-氨基丁基)苯(AM-166)阻止金黄色葡萄球菌生物膜形成和破坏生物膜的能力,方法是将龙胆紫吸附在其结构上,然后将染料解吸到有机溶剂中。金黄色葡萄球菌细胞在生物膜形成的第一阶段和成熟生物膜的组成中,使用活细胞(吖啶橙)和非活细胞(碘化丙啶)的特定染料评估其活力。采用聚合酶链反应(PCR)对琼脂糖凝胶中的PCR产物进行检测,检测抗生素耐药性和生物膜形成的相关基因。采用实时荧光定量PCR和半定量PCR检测AM-166对生物膜形成调控基因(ica、agrA、sarA和sigB)表达的影响。结果。发现化合物AM-166对金黄色葡萄球菌生物膜的形成具有抑制作用。最低抑制浓度(MIC)为5.0时,效果最显著(92.3%)。在AM-166的作用下,形成的2 d生物膜被明显破坏,在5.0 MIC时生物量下降30.9%。根据荧光显微镜的结果,5.0 MIC的金刚烷衍生物有助于减少金黄色葡萄球菌生物膜形成不同阶段的活细胞数量。分子遗传学研究结果表明,化合物AM-116的亚抑制浓度可显著抑制ica基因的表达。sarA、agrA、sigB基因的表达未见明显变化。结论。金刚烷衍生物对金黄色葡萄球菌生物膜影响的实验表明,AM-116在生物膜形成阶段的活性最明显,这可以通过抑制ica基因的转录活性来证明,ica基因负责生物膜形成的早期阶段,特别是浮游细胞对底物的粘附。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Antibiofilm Activity of 4-(Adamantyl-1)-1-(1-Aminobutyl) Benzol against Methicillin-Resistant Staphylococcus aureus
Staphylococcus aureus is a widespread opportunistic pathogen, causing community-acquired and nosocomial infections with both acute and chronic recurrent courses. The process of chronicity of the disease is provided by biofilms. Features of the structure and functioning of biofilms, in particular the presence of matrix, quorum sensing systems, persistent cells, and efflux pumps, provide microbial communities with resistance to antimicrobial drugs under their action in therapeutic concentrations. The insufficient eff ectiveness of modern antimicrobial chemotherapy against biofi lm microorganisms indicates the urgency of the problem to search for compounds with antibiofilm activity that can affect various stages of the biofilm formation and the formed biofilm. The aim of the study is to establish the antibiofilm activity of 4-(adamantyl-1)-1-(1-aminobutyl) benzol against methicillin-resistant S. aureus (MRSA) and to determine the mechanism of its action. Methods. The ability of adamantane-containing compound 4-(adamantyl-1)-1-(1-aminobutyl) benzol (AM-166) to prevent biofilm formation and destroy the formed biofilm of S. aureus was investigated on polystyrene plates by the sorption of gentian violet on its structures followed with desorption of the dye into the organic solvent. The viability of S. aureus cells at the first stage of biofilm formation and in the composition of mature biofilms was evaluated using specific dyes for living (acridine orange) and non-viable (propidium iodide) cells. Detection of genes responsible for antibiotic resistance and biofi lm formation was performed by the polymerase chain reaction (PCR) with detection of PCR products in agarose gel. Evaluation of the effect of AM-166 on the expression of genes regulating the biofilm formation (ica, agrA, sarA, and sigB) was investigated by the real-time PCR and semi-quantitative PCR. Results. It was found that the compound AM-166 shows activity against S. aureus biofilm formation. The most pronounced effect was registered at a concentration of 5.0 minimum inhibitory concentration (MIC) (92.3%.) Under the action of AM-166 on the formed 2-day biofilms, their destruction was marked: the biomass decreases by 30.9% at 5.0 MIC. According to the results of fluorescence microscopy, the adamantane derivative at 5.0 MIC helps to reduce the number of viable cells at different stages of formation of the S. aureus biofilm. The results of molecular genetic studies indicate that the ica gene expression is significantly inhibited by the action of subinhibitory concentrations of the compound AM-116. No significant changes in the expression of sarA, agrA, and sigB genes were registered. Conclusions. Experiments on the effect of adamantane derivative on S. aureus biofilms showed that the most pronounced activity of AM-116 was observed at the stage of biofilm formation, as evidenced by the inhibition of transcriptional activity of the ica gene responsible for early stages of the biofilm formation, in particular the adhesion of planktonic cells to the substrate.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Mikrobiolohichnyi zhurnal
Mikrobiolohichnyi zhurnal Medicine-Microbiology (medical)
CiteScore
0.70
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信