摘要
目的探讨邓氏清毒饮对甲型流感病毒(H3N2)的抗病毒活性及其在调节宿主炎症免疫应答中潜在的作用。方法观察邓氏清毒饮对H3N2体外活性的影响;用实时荧光定量PCR检测邓氏清毒饮对A549细胞感染H3N2病毒后白细胞介素6(IL-6)、白细胞介素8(IL-8)、干扰素诱导蛋白10(IP-10)、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)及趋化因子配体5(CCL-5)表达的影响。用蛋白印迹实验检测核因子抑制蛋白-κB(IκBα)、磷酸化核因子抑制蛋白-κB(p-IκBα)、核因子-κB p65(NF-κB p65)、磷酸化核因子-κB p65(p-NF-κB p65)的表达。结果邓氏清毒饮23.4、11.7、5.85 mg·mL^(-1)可显著抑制H3N2的体外复制,减少病毒空斑形成;在mRNA水平上可显著降低细胞因子IL-6、IL-8、IP-10、TNF-α、MCP-1及CCL-5的表达;蛋白印迹检测发现邓氏清毒饮可下调p-IκBα和p-NF-κB p65蛋白表达,增加IκBα的表达,从而抑制H3N2诱导的NF-κB信号通路激活。结论邓氏清毒饮可抑制H3N2的体外复制,抑制流感病毒诱导的NF-κB信号通路激活,降低炎性细胞因子的表达,表明邓氏清毒饮具有防治流感病毒的潜力。
Objective This study was designed to investigate the antiviral activity of Dengshi Qingdu Decoction against influenza virus(H3 N2)and its potential role in regulating inflammatory host immune response.MethodsThe in vitro inhibitory effect of Dengshi Qingdu Decoction on the replication of influenza virus(H3 N2)was evaluated by plaque reduction and other tests.Real-time RT-PCR was used to detect the expression levels of pro-inflammatory cytokines(IL-6,IL-8,IP-10,TNF-α,MCP-1 and CCL-5)in A549 cells infected with influenza virus(H3 N2).Western Blot was used to detect the expression levels of IκBα,p-IκBα,NF-κB p65,p-NF-κB p65.Results The results showed that 23.4,11.7,5.85 mg·mL-1 Dengshi Qingdu Decoction could significantly inhibit the replication of H3 N2 in vitro,reduce the formation of viral plaque.Real-time RT-PCR showed that Dengshi Qingdu Decoction could significantly reduce the mRNA levels of IL-6,IL-8,IP-10,TNF-α,MCP-1 and CCL-5 in the infected cells.Western Blot detection showed that Dengshi Qingdu Decoction could significantly reduce the expression of p-IκBαand p-NF-κB p65,while increase the IκBαin the infected cells,thus,inhibit the activation of NF-κB signaling pathway induced by influenza virus(H3 N2),and reduce the expression of inflammatory cytokines.ConclusionStudies have shown that Dengshi Qingdu Decoction can inhibit the replication of influenza virus(H3 N2)in vitro,inhibit the activation of NF-κB signal pathway induced by influenza virus,and reduce the expression of inflammatory cytokines,which proves that Dengshi Qingdu decoction has the potential of preventing and treating influenza virus.
作者
马钦海
陈瑞晗
雷标
王周琅
朱梓宁
黄子天
陈凯佳
邱仕君
刘小斌
邓中光
杨子峰
MA Qinhai;CHEN Ruihan;LEI Biao;WANG Zhoulang;ZHU Zining;HUANG Zitian;CHEN Kaijia;QIU Shijun;LIU Xiaobin;DENG Zhongguang;YANG Zifeng(State Key Laboratory of Respiratory Disease,National Clinical Research Center for Respiratory Disease,Guangzhou Institute of Respiratory Health,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 511436 Guangdong,China;Denglao JinFang Pharmaceutical Technology Co.,Ltd.,Meizhou 514000 Guangdong,China;School of Basic Medicine,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405 Guangdong,China;Guangdong New South China Academy of Traditional Chinese Medicine,Guangzhou 510230 Guangdong,China;State Key Laboratory of Quality Research in Chinese Medicine,Macao Institute for Applied Research in Medicine and Health,Macao University of Science and Technology,Taipa 519020 Macao SAR,China;KingMed Virology Diagnostic&Translational Center,Guangzhou 510725 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第5期597-603,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家重点研发计划项目(2017YFC1309302)
国家自然科学基金面上项目(81873065)
澳门科学技术发展基金项目(0172/2019/A3)
广州医科大学高水平大学建设项目([2017]159号、160号)。
