基于蛋白质组学探讨雾化吸入BD-77对呼吸道病毒感染的抑制作用及机制  

Preliminary Proteomics-based Investigation of Inhibitory Effect and Mechanism of BD-77 by Nebulized Inhalation on Respiratory Viral Infections

作　　者：包蕾[1] 耿子涵 郭姗姗[1] 周利润 赵荣华[1] 孙静[1] 鲍岩岩[1] 李星 黄慈钢 蒋坤 彭飞燕 徐洲 黄成钢 崔晓兰[1] 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)

机构地区：[1]中国中医科学院中药研究所,北京100700 [2]贵州百灵企业集团制药股份有限公司,贵州安顺561099 [3]中国科学院上海药物研究所,上海201203

出　　处：《中国实验方剂学杂志》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)。

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 sig

关 键 词：呼吸道病毒 雾化吸入法 BD-77 蛋白质组学 肺部炎症 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33