摘要
目的:观察雾化吸入BD-77对多种呼吸道病毒感染动物模型的治疗作用,并利用蛋白质组学手段,探究BD-77广谱抗病毒作用机制。方法:流感病毒H1N1/FM1实验选用ICR小鼠分为正常组、模型组、达菲组、75、37.5 g·L^(-1)均分别吸入20 min组、25 min组;人冠状病毒229E和人冠状病毒OC43实验选用BALB/c小鼠正常组、模型组、磷酸氯喹组、BD-77的75、37.5、18.75、9.375 g·L^(-1)剂量组,每组10只,分别采用流感病毒H1N1/FM1、人冠状病毒229E和人冠状病毒OC43感染致肺炎模型,检测小鼠肺指数、实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织病毒载量、酶联免疫吸附测定法(ELISA)检测肺组织中相关炎性因子,并对OC43感染小鼠肺组织进行蛋白质组学分析。结果:与正常组比较,各感染组小鼠肺指数显著升高(P<0.01)、人冠状病毒229E和人冠状病毒OC43小鼠肺组织内能检测到病毒核酸,且人冠状病毒229E感染小鼠肺组织中白细胞介素-6(IL-6)、IL-10、肿瘤坏死因子-α(TNF-α)含量均显著升高(P<0.01);BD-77能够明显降低流感病毒H1N1/FM1、人冠状病毒229E和人冠状病毒OC43感染小鼠的肺指数(P<0.05,P<0.01);显著降低人冠状病毒229E和人冠状病毒OC43感染小鼠的肺内病毒载量(P<0.01);显著降低人冠状病毒229E感染小鼠肺组织中IL-6、IL-10、TNF-α的含量(P<0.01)。OC43感染小鼠肺组织蛋白质组学显示,BD-77调节了腺苷酸活化蛋白激酶(AMPK)信号通路、TNF信号通路、NOD样(NOD-like)信号通路、IL-17信号通路、叉头框蛋白O(FoxO)信号通路和转化生长因子-β(TGF-β)信号通路等。结论:雾化吸入BD-77能够有效治疗流感病毒H1N1/FM1、人冠状病毒229E和人冠状病毒OC43感染小鼠所致肺炎,并且可能是通过调节细胞代谢平衡、增强宿主免疫功能、减轻炎症反应来发挥抗病毒作用的。
Objective:To observe the therapeutic effect of BD-77 by nebulized inhalation on animal models of various respiratory viral infections and investigate the mechanism of broad-spectrum antiviral action of BD-77 using proteomics.Method:The influenza virus H1N1/FM1 experiment used ICR mice and divided them into a normal group,model group,Tamiflu group,and BD-77 groups of 75 and 37.5 g·L^(-1) for inhalation of 20 min and 25 min.Human coronavirus 229E and OC43 experiment divided the BALB/c mice into a normal group,model group,chloroquine phosphate group,and BD-77 groups of 75,37.5,18.75,and 9.375 g·L^(-1),with 10 mice in each group.Influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infectioninduced pneumonia models were used to detect mouse lung index,and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the viral load in lung tissue.Enzyme-linked immunosorbent assay(ELISA)was used to detect related inflammatory factors in lung tissue,and proteomics analysis was performed on the lung tissue of OC43-infected mice.Result:Compared with that in the normal group,the lung index of mice in each infection group was significantly increased(P<0.01),and viral nucleic acid could be detected in the lung tissue of mice infected with human coronaviruses 229E and OC43.The levels of interleukin-6(IL-6),IL-10,and tumor necrosis factor-α(TNF-α)in the lung tissue of mice infected with human coronavirus 229E were all significantly increased(P<0.01).BD-77 could significantly reduce the lung index of mice infected with influenza virus H1N1/FM1 and human coronaviruses 229E and OC43(P<0.05,P<0.01),cut down the viral load in the lungs of mice infected with human coronaviruses 229E and OC43(P<0.01),and lower the contents of IL-6,IL-10,and TNF-αin the lung tissue of mice infected with human coronavirus 229E(P<0.01).Proteomics analysis of the lung tissue of OC43-infected mice showed that BD-77 regulated the AMPK signaling pathway,TNF signaling pathway,NOD-like signaling pathway,IL-17 signaling pathway,Forkhead box protein O(FoxO)signaling pathway,transforming growth factor-β(TGF-β)signaling pathway,and other signaling pathways.Conclusion:Nebulized inhalation of BD-77 is effective in treating pneumonia caused by influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection in mice and may exert its antiviral effects by regulating the balance of cellular metabolism,enhancing the immune function of the host,and attenuating inflammatory responses.
作者
包蕾
耿子涵
郭姗姗
周利润
赵荣华
孙静
鲍岩岩
李星
黄慈钢
蒋坤
彭飞燕
徐洲
黄成钢
崔晓兰
BAO Lei;GENG Zihan;GUO Shanshan;ZHOU Lirun;ZHAO Ronghua;SUN Jing;BAO Yanyan;LI Xing;HUANG Cigang;JIANG Kun;PENG Feiyan;XU Zhou;HUANG Chenggang;CUI Xiaolan(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Medical Sciences Guizhou Bailing Enterprise Group Pharmaceutical Co.Ltd,Anshun 561099,China;Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第13期52-59,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82141206,82151210)。
