木质素抑制金黄色葡萄球菌所致皮肤感染的机制研究:生物膜形成核心基因筛选与验证
Mechanism of Lignin Inhibition of Skin Infections Caused by Staphylococcus au-reus:Screening and Validation of Hub Genes for Biofilm Formation
黄树强 1谭翠钰 1陈淼琪 1袁晓珺 1李志宏 1陈婉茹 2李芝鸿 1张彤彤 1徐令清1
作者信息
- 1. 广州医科大学附属清远医院(清远市人民医院)检验科,清远,511518
- 2. 广州医科大学第三临床学院,广州,511436
- 折叠
摘要
金黄色葡萄球菌(Staphylococcus aureus,SA)生物膜形成常导致严重的皮肤感染.木质素是抑制SA生长的有效药物,分析木质素对SA生物膜形成的抑制作用和分子机制有助于为临床治疗皮肤感染提供指导.本研究通过结晶紫染色实验证实木质素对SA生物膜的形成有抑制作用.随后,基于公共数据库获取木质素抑制SA生物膜形成的相关基因,结合机器学习与分子对接方法筛选核心基因.同时,设计人体皮肤SA生长密度预测模型,分析人体皮肤SA生长密度与生物膜形成相关基因表达水平的相关性.最后,通过实时定量PCR(real-time quantitative PCR,RT-qPCR)实验验证核心基因的表达水平.结果显示,木质素最小抑菌浓度(minimum inhibitory concentration,MIC)为16 mg/mL.随着木质素浓度升高,SA生物膜产量逐步减少.purA、trpB、hla、uhpT、leuC等核心基因拥有优越的与木质素结合的性能.人体皮肤预测模型显示,核心基因中trpB、leuC、leuD、dnaI与SA的皮肤生长密度评分呈负相关,而uhpT、hla、purA、dnaN与评分呈正相关.RT-qPCR结果表明,与未经处理的对照菌株相比,木质素处理后的菌株中hla、purA和uhpT基因的转录水平显著下降,而teuC和trpB基因的转录水平显著上升,与数据分析结果一致.本研究揭示了木质素对SA生物膜形成的靶向抑制作用,并阐明了其在SA皮肤感染中的治疗价值.
Abstract
Staphylococcus aureus(SA)biofilm formation frequently leads to severe skin infections.Lignin is an effective drug to inhibit the growth of SA,and analyzing the inhibitory effect and molecular mechanism of lignin on SA biofilm formation contributes to providing guidance for clinical treatment of skin infections.In this study,inhibition of SA biofilm formation by lignin was confirmed by crystal violet staining assay.Subsequently,we obtained the relevant genes of lignin inhibiting SA biofilm formation based on the public data-base and screened the hub genes by combining machine learning and molecular docking methods.At the same time,a human skin SA growth density prediction model was designed to analyze the correlation between human skin SA growth density and the expression levels of genes related to biofilm formation.Finally,the expression levels of hub genes were verified by real-time quantitative PCR(RT-qPCR).The results showed that the minimum inhibitory concentration(MIC)of lignin was 16 mg/mL.Biofilm production de-creased progressively with increasing lignin concentrations.The hub genes pur A,trpB,hla,uhpT,and leuC possessed superior binding properties with lignin.The human skin prediction model showed that the hub genes trpB,leuC,leuD,and dnaI were negatively corre-lated with density scores of SA,whereas the hub genes uhpT,hla,pur A,and dnaN were positively correlated with density scores of SA.RT-qPCR results showed that the transcript levels of hla,purA and uhpT genes were significantly decreased in the lignin-treated strain compared with the untreated control strain,whereas the transcript levels of leuC and trpB genes were significantly increased,which was consistent with the results of the data analysis.This study reveals the targeted inhibitory effect of lignin on SA biofilm forma-tion and elucidates its therapeutic value in SA skin infections.
关键词
木质素/金黄色葡萄球菌/生物膜/皮肤感染/生物信息学Key words
Lignin/Staphylococcus aureus/Biofilm/Skin infection/Bioinformatics引用本文复制引用
基金项目
广东省钟南山医学基金会资助科研项目(ZNSXS-20240010)
广东省医学科学技术研究基金资助项目(A2021490)
清远市人民医院医学科研基金资助项目(202301-201)
清远市人民医院医学科研基金资助项目(20190209)
2023年度广州医科大学学生创新能力提升计划资助项目(02-408-240603131130)
出版年
2024
NETL
NSTL
万方数据