摘要
目的探究中药勒马回抑制耐甲氧西林金黄色葡萄球菌(MRSA)耐药生物膜的作用和机制。方法培养MRSA至耐药生物膜成熟后,给予勒马回注射液处理,用扫描电子显微镜(SEM)观察生物膜超微结构。基于文献和数据库检索获得勒马回主要活性成分和生物膜耐药性相关基因,用Swiss Targets Predictio、UniProt数据库获得有效成分的3D结构和生物膜耐药性相关靶点,用Cytoscape软件构建勒马回的活性成分、靶点的关系网络。利用STRING构建蛋白质-蛋白质相互作用(PPI)网络;用AutoDock Vina软件,将关键活性成分与核心靶点进行分子对接,并用Gromacs2022对关键活性成分与核心靶点进行100 ns分子动力学模拟,筛选有效成分和基因;用反转录定量聚合酶链式反应(RT-qPCR)验证勒马回及其主要成分对耐药生物膜形成及耐药相关核心基因的影响。结果勒马回对MRSA生物膜形成有抑制作用;其主要成分原儿茶酸(PCA)和水杨酸(SA)可与MRSA耐药生物膜核心靶点SarA、SarX、Rot、SigB紧密结合,结合能均低于-4.5 kcal/mol;分子动力学模拟结果表明,勒马回复合物整体结构稳定;RT-PCR显示,勒马回能显著下调核心调控基因的表达。结论勒马回的活性成分PCA和SA可能通过多靶点协同干扰生物膜相关调控网络、下调关键调控基因表达来抑制MRSA耐药生物膜的形成。
Objective To investigate the inhibitory effects and underlying mechanisms of the Chinese medicinal material Veronica linariifolia(Lemahui,LMH)on methicillin-resistant Staphylococcus aureus(MRSA)drug-resistant biofilms.Methods MRSA was cultured to form mature drug-resistant biofilms,which were then treated with LMH injection.The ultrastructure of the biofilms was observed using scanning electron microscopy(SEM).Based on literature review and database retrieval,the main active components of LMH and genes associated with biofilm drug resistance were identified.The 3D structures of the active compounds and the corresponding biofilm drug-resistance targets were obtained from the SwissTargetPrediction and UniProt databases.Cytoscape software was used to construct the component-target interaction network of LMH.A protein-protein interaction(PPI)network was established using the STRING database.Molecular docking between key active compounds and core targets was performed using AutoDock Vina,followed by 100 ns molecular dynamics(MD)simulations with Gromacs 2022 to screen effective components and genes.Finally,reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to validate the effects of LMH and its major active components on the expression of core genes related to biofilm formation and drug resistance.Results LMH inhibited MRSA biofilm formation.Its main constituents,protocatechuic acid(PCA)and salicylic acid(SA),bound tightly to the core targets of MRSA drug-resistant biofilms SarA,SarX,Rot,and SigB,with binding energies below−4.5 kcal/mol.Molecular dynamics simulations indicated overall structural stability of these complexes.RT-qPCR showed that LMH significantly downregulated the expression of these core regulatory genes.Conclusion LMH inhibits the formation of MRSA drug-resistant biofilms.Its mechanisms may involve multi-target synergistic modulation by PCA and SA,which interfere with biofilm-associated regulatory networks and downregulate the expression of key regulatory genes.
作者
安洪萨
戎新倩
杨伟峰
张璐璐
郑昊然
陈艺幻
姜春燕
吕诚
谭勇
AN Hongsa;RONG Xinqian;YANG Weifeng;ZHANG Lulu;ZHENG Haoran;CHEN Yihuan;JIANG Chunyan;LYU Cheng;TAN Yong(Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Medical Experimental Center,China Academy of Chinese Medical Sciences,Beijing 100700;Department of Dermatology,Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010)
出处
《北京中医药》
2025年第10期1285-1291,共7页
Beijing Journal of Traditional Chinese Medicine
基金
中国中医科学院科技创新工程中医临床基础学科创新团队项目(C2021B003)
中国中医科学院自主选题项目(Z0735)。
关键词
勒马回
耐甲氧西林金黄色葡萄球菌
耐药生物膜
网络药理学
分子动力学
Veronica linariifolia
methicillin-resistant Staphylococcus aureus
drug-resistant biofilm
network pharmacology
molecular dynamics
