北豆根总生物碱的抗病毒感染功能与机制  

Effect and Mechanism of Menispermi Rhizoma Total Alkaloids on Antiviral Infection

作　　者：王雪娇 李琪琪 禹艳丽 刘霞 李敏 刘哲 贾鑫 王遥 WANG Xuejiao;LI Qiqi;YU Yanli;LIU Xia;LI Min;LIU Zhe;JIA Xin;WANG Yao(School of Life Science,Beijing University of Chinese Medicine,Beijing 100027,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100027,China)

机构地区：[1]北京中医药大学生命科学学院,北京100027 [2]北京中医药大学中药学院,北京100027

出　　处：《中国实验方剂学杂志》2024年第10期37-44,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市科技新星计划项目(2023107);中华中医药学会青年人才托举工程项目A类资助项目;国家自然科学基金项目(82001663);北京市自然科学基金项目(7242239)。

摘　　要：目的:探讨北豆根总生物碱的抗病毒感染作用及其与Ⅰ型干扰素(IFN-Ⅰ)信号通路的关系。方法:通过倒置荧光显微镜、流式细胞术、实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)等方法检测北豆根总生物碱对甲型流感病毒(H1N1)、水疱性口炎病毒(VSV)和脑心肌炎病毒(EMCV)在细胞内复制的影响。构建H1N1病毒感染小鼠模型,将小鼠分为空白组、H1N1模型组、北豆根总生物碱组(10、20、30 mg·kg^(-1))和奥司他韦组(40 mg·kg^(-1)),研究北豆根总生物碱对感染小鼠的体质量和生存率的影响。Real-time PCR检测北豆根总生物碱对细胞中IFN-Ⅰ通路的激活作用,并在干扰素受体1(IFNAR1)敲除的A549细胞(IFNAR1^(-/-)-A549)检测北豆根总生物碱抗病毒感染作用与IFN-Ⅰ通路的关系。结果:与空白组比较,病毒感染模型组中的病毒大量增殖(P<0.01);与模型组比较,北豆根总生物碱在体外显著抑制H1N1、VSV和EMCV的复制(P<0.01),在体内抑制H1N1病毒感染小鼠体质量降低,提升小鼠生存率(P<0.05)。此外,北豆根总生物碱激活IFN-Ⅰ通路并依赖该通路发挥抗病毒感染作用。结论:北豆根总生物碱通过激活IFN-Ⅰ通路发挥体内和体外的抗病毒感染作用。Objective:To investigate the antiviral effect of Menispermi Rhizoma total alkaloids and its relationship with the typeⅠinterferon(IFN-Ⅰ)signaling pathway.Method:The effects of Menispermi Rhizoma total alkaloids on the intracellular replication of influenza A virus(H1N1),vesicular stomatitis virus(VSV),and cerebral myocarditis virus(EMCV)were detected by fluorescent inverted microscope,flow cytometry,Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and Western blot.A mouse model infected with H1N1 was constructed,and the mice were divided into a control group,H1N1 model group,Menispermi Rhizoma total alkaloids groups(10,20,30 mg·kg^(-1)),and oseltamivir group(40 mg·kg^(-1)),so as to study the effects on the weight and survival rate of infected mice.Real-time PCR was used to detect the activation effect of Menispermi Rhizoma total alkaloids on the IFN-Ⅰ pathway in cells,and the relationship between the antiviral effect of Menispermi Rhizoma total alkaloids in IFNAR1 knockout A549 cells(IFNAR1^(-/-)-A549)and IFN-Ⅰ pathway was detected.Result:Compared with the control group,the virus proliferated significantly in the model group(P<0.01).Compared with the model group,Menispermi Rhizoma total alkaloids could significantly inhibit the replication of H1N1,VSV,and EMCV in vitro(P<0.01),inhibit the weight loss of the mice infected with the H1N1 in vivo,and improve the survival rate of mice(P<0.05).In addition,Menispermi Rhizoma total alkaloids activated the IFN-Ⅰ pathway and relied on this pathway to exert the function of antiviral infection.Conclusion:Menispermi Rhizoma total alkaloids exert antiviral effects in vivo and in vitro by activating the IFN-Ⅰ pathway.

关 键 词：北豆根总生物碱 甲型流感病毒(H1N1) 水疱性口炎病毒(VSV) 脑心肌炎病毒(EMCV) 抗病毒免疫 Ⅰ型干扰素(IFN-Ⅰ) 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33