摘要
目的探索连翘水提液中与RSV-F蛋白特异性结合的效应物质,并验证其作用效果。方法采用表面等离子共振亲和垂钓技术,富集、再生和回收连翘水提液中与RSV-F蛋白特异性结合的活性成分,运用超高效液质联用分析对回收样本进行分析鉴定,并采用分子对接、分子动力学模拟及SPR亲和力检测筛选与F蛋白稳定结合的活性成分,利用动物实验进行药效学验证。将BALB/c小鼠随机分为正常组、模型组、阳性对照组、原儿茶酸组和毛蕊花糖苷组。实验组均滴鼻RSV造模;各给药组于造模当天开始灌胃干预,连续4 d后取材。采用HE染色观察肺组织病变情况,ELISA法检测肺组织炎性因子变化。结果多种连翘水提液活性成分可与RSV-F蛋白结合,其中以毛蕊花糖苷、原儿茶酸结果更为稳定;体内药效学结果表明,与正常组相比,模型组小鼠肺组织有明显病变,炎性因子水平升高,采用毛蕊花糖苷、原儿茶酸干预后病变状态有所改善。结论连翘可通过毛蕊花糖苷、原儿茶酸等多种连翘水提液活性成分干预RSV-F蛋白表达以减轻RSV引起的肺组织病变程度,可能是其抗RSV的物质基础。
Objective To explore the effective substances that specifically bind to RSV-F protein in the water extract of Forsythia suspensa,and verify its effect.Methods Surface plasmon resonance(SPR)affinity fishing technology was used to enrich,regenerate and recover the active components that specifically bind to RSV-F protein in the water extract of Forsythia suspensa.The recovered samples were analyzed and identified by UHPLC-MS/MS,and the active components that stably bind to F protein were screened by molecular docking,molecular dynamics simulation and SPR affinity detection,and then returned to animal experiments for pharmacodynamics verification.BALB/c mice were randomly divided into normal group,model group,positive control group,protocatechuic acid group and verbascoside group.In the experimental group,RSV was dripped into the nose to make models.Each drug group began to be given intragastric intervention on the day of modeling,and samples were taken after 4 days.HE staining was used to observe the pathological changes of lung tissue,and ELISA was used to detect the changes of inflammatory factors in lung tissue.Results A variety of active components in the aqueous extract of Forsythia suspensa could bind to RSV-F protein,among which the results of verbascoside and protocatechuic acid were more stable.In vivo pharmacodynamics experiments showed that the lung tissue of the model group had obvious pathological changes and the level of inflammatory factors increased,and the pathological state was improved after the intervention of verbascoside and protocatechuic acid compared with the normal group.Conclusion Forsythia suspensa can interfere with the expression of RSV-F protein through many active components such as verbascoside and protocatechuic acid,which may be the material basis of its anti-RSV.
作者
王琳
王平
蔡雯
丁晓彦
王仪
王成
张梦茹
杜海涛
WANG Lin;WANG Ping;CAI Wen;DING Xiaoyan;WANG Yi;WANG Cheng;ZHANG Mengru;DU Haitao(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Shandong Academy of Chinese Medicine,Jinan 250014,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第5期1405-1416,共12页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
山东大学科学技术局山东大学“新高校20条”资助项目(202228121):山东道地中药材抗病毒研究科研带头人工作室,负责人:王平
山东省中医药管理局山东省中医药科技项目(Q-2023023):基于决策树预测模型的连翘靶向呼吸道合胞病毒F蛋白效应物质筛选,负责人:杜海涛
山东省科学技术厅山东省重点研发计划(科技示范工程)(2021SFGC1205):山东省重点研发计划(科技示范工程)药食同源特色产品创新研发与应用示范项目,负责人:王平。
关键词
连翘
SPR垂钓技术
RSV-F蛋白
分子对接
分子动力学模拟
Forsythia suspensa
SPR fishing technology
RSV-F protein
Molecular docking
Molecular dynamics simulation
