月季花活性成分槲皮素抗烟曲霉的宿主-病原体通路互作机制研究

doi:10.3870/j.issn.1672-8009.2026.03.005

摘要/Abstract

摘要： 目的基于宿主-病原体互作视角,探讨月季花的主要活性成分槲皮素抗烟曲霉的多靶点作用机制。方法利用TCMSP、GeneCards和OMIM数据库筛选月季花活性成分及烟曲霉感染相关宿主靶点;从NCBI和UniProt数据库获取烟曲霉毒力及耐药相关靶点;借助KEGG Orthology系统进行跨物种同源映射,运用Cytoscape 3.10.3构建宿主-病原体通路互作网络(host-pathogen pathway interaction network,HPPIN),通过拓扑分析(度中心性、介数中心性)识别核心靶点。采用AutoDock Vina1.5.7软件对槲皮素与核心靶点进行分子对接,结合能阈值≤-5.0 kcal/mol;选取最优复合物,使用GROMACS 2020.6软件进行100 ns分子动力学模拟(AMBER99SB力场),分析均方根偏差(RMSD)、均方根涨落(RMSF)及氢键数量以评估结合稳定性。最后参照CLSI M38-A3方案,采用微量液基稀释法测定槲皮素对烟曲霉标准株Af293的最低抑菌浓度(MIC);通过N-乙酰半胱氨酸和嘌呤混合物体外表型拯救实验,验证预测的氧化应激与嘌呤代谢干扰两条作用路径,倒置显微镜观察菌丝形态变化。结果 HPPIN识别出12个关键靶点。槲皮素与KRas及SAICAR合成酶的结合能分别为-7.6与-7.8 kcal/mol,复合物稳定(RMSD<0.2 nm)。其对烟曲霉MIC为256 μg/mL,完全抑制菌丝生长。此效应可被N-乙酰半胱氨酸逆转,证实氧化应激机制;与高浓度嘌呤联用导致萌发完全失败,揭示嘌呤代谢紊乱。结论槲皮素通过“断供”与“下毒”双路径协同抑制烟曲霉。

关键词: 宿主-病原体互作, 烟曲霉, 网络药理学, 分子对接, 分子动力学, 槲皮素, 月季花

Abstract: Objective To investigate the multi-target mechanism of quercetin,the main active component of Rosa chinensis(R.chinensis),against Aspergillus fumigatus(A.fumigatus) from the host-pathogen interaction perspective. Methods Firstly,the active components of R.chinensis and host targets related to A.fumigatus infection were screened using TCMSP,GeneCards,and OMIM databases.Virulence and drug resistance-related targets of A.fumigatus were obtained from NCBI and UniProt databases.Cross-species homology mapping was performed using the KEGG Orthology system,and a Host-Pathogen Pathway Interaction Network(HPPIN)was constructed using Cytoscape 3.10.3.Core targets were identified through topological analysis(degree centrality and betweenness centrality).Secondly,molecular docking of quercetin with the core targets was conducted using AutoDock Vina 1.5.7,with a binding energy threshold of ≤ -5.0 kcal/mol.The optimal complex was selected for a 100 ns molecular dynamics simulation using GROMACS 2020.6(AMBER99SB force field)to analyze binding stability via root-mean-square deviation(RMSD),root-mean-square fluctuation(RMSF),and hydrogen bond numbers.Finally,following the CLSI M38-A3 protocol,the minimum inhibitory concentration(MIC)of quercetin against the standard A.fumigatus strain Af293 was determined using the broth microdilution method.Phenotypic rescue experiments using a mixture of N-acetylcysteine and purine were performed to validate the predicted interference with oxidative stress and purine metabolism pathways,and hyphal morphological changes were observed under an inverted microscope. Results The HPPIN identified 12 key targets.Quercetin exhibited binding energies of-7.6 kcal/mol with KRas and-7.8 kcal/mol with SAICAR synthetase,forming stable complexes(RMSD<0.2 nm).The MIC of quercetin against A.fumigatus was 256 μg/mL,resulting in complete inhibition of hyphal growth.This inhibitory effect was reversed by N-acetylcysteine,confirming the role of oxidative stress.Concurrently,combination with high-concentration purine led to complete germination failure,indicating disruption of purine metabolism. Conclusion Quercetin synergistically inhibits A.fumigatus through dual mechanisms of “starvation” and “intoxication.”

Key words: host-pathogen interaction, Aspergillus fumigatus, network pharmacology, molecular docking, molecular dynamics, quercetin, Rosa chinensis

中图分类号:

R379

高小喻, 马彦. 月季花活性成分槲皮素抗烟曲霉的宿主-病原体通路互作机制研究[J]. 医学分子生物学杂志, 2026, 23(3): 266-274.

GAO Xiaoyu, MA Yan. Mechanisms of Host-Pathogen Pathway Interactions Mediated by Quercetin Against Aspergillus fumigatus[J]. Journal of Medical Molecular Biology, 2026, 23(3): 266-274.

参考文献

[1] LATGÉ J P,CHAMILOS G.Aspergillus fumigatus and aspergillosis in 2019[J].Clin Microbiol Rev,2019,33(1):e00140-e00118.
[2] HOSSEINI P,KENIYA M V,SAGATOVA A A,et al.The molecular basis of the intrinsic and acquired resistance to azole antifungals in Aspergillus fumigatus[J].J Fungi(Basel),2024,10(12):820.
[3] VAN DE VEERDONK F L,GRESNIGT M S,ROMANI L,et al.Aspergillus fumigatus morphology and dynamic host interactions[J].Nat Rev Microbiol,2017,15(11):661-674.
[4] 阿里木·艾山,王晓东.唑类耐药烟曲霉的耐药机制[J].中国真菌学杂志,2025,20(1):85-91.
Alimu A,WANG X D.Drug resistance mechanism of azole-resistant Aspergillus fumigatus[J].Chin J Mycol,2025,20(1):85-91.
[5] IVANA D R,MILAN S S,OLGICA D S,et al.Anti-Aspergillus properties of different extracts from selected plants[J].Afr J Microbiol Res,2011,5(23):3986-3990.
[6] HE J,DONG J,ZHANG W,et al.Comparative unravelling the antifungal activity and mechanism of oleuropein and matrine inhibiting the growth of phytopathogen Fusariumspp[J].Pestic Biochem Physiol,2025,213:106538.
[7] 靳蕊,徐敏纹,刘莹,等.黄酮类化合物的抑菌作用及其机制的研究[J].继续医学教育,2016,30(8):152-154.
JIN R,XU M W,LIU Y,et al.Study on bacteriostasis of flavonoids and its mechanism[J].Continuing Med Educ,2016,30(8):152-154.
[8] 吴晶,王亮,曹永兵,等.《中药大辞典》记载的抗真菌中药体外药效再评价[J].中国真菌学杂志,2016,11(6):348-353.
WU J,WANG L,CAO Y B,et al.Re-evaluation of the traditional Chinese medicines(TCMs)with antifungal activities recorded in‘Zhong Yao Da Ci Dian’[J].Chin J Mycol,2016,11(6):348-353.
[9] GARCÍA CARNERO L C,LI X,DE CASTRO P A,et al.Colistin enhances caspofungin antifungal efficacy against Aspergillus fumigatus by modulating calcium homeostasis and stress responses[J].Nat Commun,2025,16(1):5967.
[10] ZHANG G B,LI Q Y,CHEN Q L,et al.Network pharmacology:a new approach for Chinese herbal medicine research[J].Evid Based Complement Alternat Med,2013,2013:621423.
[11] CHEN L,DENG H,CUI H,et al.Inflammatory responses and inflammation-associated diseases in organs[J].Oncotarget,2018,9(6):7204-7218.
[12] KANEHISA M,FURUMICHI M,SATO Y,et al.KEGG:biological systems database as a model of the real world[J].Nucleic Acids Res,2025,53(D1):D672-D677.
[13] TROTT O,OLSON A J.AutoDock Vina:Improving the speed and accuracy of docking with a new scoring function,efficient optimization,and multithreading[J].J Comput Chem,2010,31(2):455-461.
[14] ABRAHAM M J,MURTOLA T,SCHULZ R,et al.GROMACS:High performance molecular simulations through multi-level parallelism from laptops to supercomputers[J].SoftwareX,2015,1-2:19-25.
[15] LINDORFF-LARSEN K,PIANA S,PALMO K,et al.Improved side-chain torsion potentials for the Amber ff99 SB protein force field[J].Proteins,2010,78(8):1950-1958.
[16] KAWASAKI L,WYSONG D,DIAMOND R,et al.Two divergent catalase genes are differentially regulated during Aspergillus nidulans development and oxidative stress[J].J Bacteriol,1997,179(10):3284-3292.
[17] NIERMAN W C,PAIN A,ANDERSON M J,et al.Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus[J].Nature,2005,438(7071):1151-1156.
[18] SNELDERS E,CELIA-SANCHEZ B N,NEDERLOF Y C,et al.Widely dispersed clonal expansion of multi-fungicide-resistant Aspergillus fumigatus limits genomic epidemiology prospects[J].mBio,2025,16(6):e0365224.
[19] YIN J,PENG X,LIN J,et al.Quercetin ameliorates Aspergillus fumigatus keratitis by inhibiting fungal growth,toll-like receptors and inflammatory cytokines[J].Int Immunopharmacol,2021,93:107435.
[20] CHEN F,ZHANG X,GUO R,et al.Glycolysis-mediated energy metabolism mechanisms:Synergistic antifungal activity of(±)-methylagrimonolide against drug-resistantCandidaalbicans[J].Bioorg Chem,2025,167:109253.
[21] ZHOU M,LIU L,CONG Z,et al.Author Correction:A dual-targeting antifungal is effective against multidrug-resistant human fungal pathogens[J].Nat Microbiol,2024,9(5):1394.