银花平感颗粒和三拗汤体内体外结合对甲型流感病毒的影响  

Effects of Yinhua Pinggan Granule and San-ao Decoction on Influenza A Virusin Vitro and in Vivo

作　　者：杜海霞 周惠芬 何昱[3] 尹斐 杨洁红 卢亦愚[4] 万海同[1,2] DU Hai-xia;ZHOU Hui-fen;HE Yu;YIN Fei;YANG Jie-hong;LU Yi-yu;WAN Hai-tong(School of Basic Medical Science,Zhejiang Chinese Medicine University,Hangzhou(310053 );College of Life Sciences,Zhejiang Chinese Medical University,Hangzhou (310053);College of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou (310053);Institute of Microbiology,Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou(310009 ))

机构地区：[1]浙江中医药大学基础医学院,杭州310053 [2]浙江中医药大学生命科学学院,杭州310053 [3]浙江中医药大学药学院,杭州310053 [4]浙江省疾病预防控制中心微生物所,杭州310009

出　　处：《中国中西医结合杂志》2019年第2期223-229,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81603531);浙江省自然科学基金资助项目(No.LZ14H270001);浙江省卫生领军人才培养项目(No.浙卫发[2016]6号)

摘　　要：目的通过银花平感颗粒和三拗汤两方药体内外对甲型H1N1流感病毒影响的实验研究,探讨不同方剂配伍对流感病毒的抑制作用。方法体内实验:采用滴鼻感染建立小鼠流感病毒肺炎模型,按体重分层随机分为正常对照组、病毒模型组、达菲组[21. 63 mg/(kg·d)]、银花平感颗粒组[12 g/(kg·d)]和三拗汤组[9 g/(kg·d)],灌胃给予相应药物,每天1次,连续5天。灌胃结束后,摘取肺、脾及胸腺组织,分别称重,计算各组小鼠肺指数、肺组织抑制率及免疫脏器指数;采用实时荧光定量PCR法检测各组小鼠肺组织病毒载量表达,苏木精-伊红(HE)染色法检测各组小鼠肺组织的病理改变。体外实验:将银花平感颗粒和三拗汤两方药作用于感染甲型H1N1流感病毒的犬肾细胞(MDCK),采用MTT法检测两方药对甲型H1N1流感病毒的抗病毒有效率及显微镜观察其引起的细胞病变效应(CPE)。结果体内实验中,与正常对照组比较,病毒模型组小鼠肺指数升高,脾指数、胸腺指数均降低,多处肺实变,炎症细胞浸润,肺组织内病毒载量高表达(P <0. 01),与病毒模型组比较,银花平感颗粒和三拗汤均降低流感病毒感染小鼠肺指数(P <0. 01),升高脾指数、胸腺指数(P <0. 05,P <0. 01),并明显减轻流感病毒肺炎小鼠肺组织病理变化,降低肺组织病毒载量(P <0. 05,P <0. 01);体外实验中,银花平感颗粒和三拗汤可以明显抑制流感病毒引起的CPE,对病毒感染也有明显的保护作用。结论银花平感颗粒和三拗汤两方药在体内外均有良好的病毒抑制作用,其抗病毒作用可能与抑制流感病毒复制、保护免疫器官、减轻肺部炎症损伤及抑制病毒引起的CPE有关,且在体内外对流感病毒的抑制方面,银花平感颗粒比三拗汤配伍更全面,疗效更好。Objective To evaluate the effects of Yinhua Pinggan Granule and San-ao Decoction on inhibition influenza A/H1 N1 virusin vitro andin vivo,and to investigate the inhibitory effect of different compatibility of formula on the influenza A/H1 N1 virus. Methods In vivo experiments,the model of pneumonia by nasal dropping influenza virus was established in mice. According to the weight stratification,then they were divided randomly into five groups: normal control group,influenza virus model group,oseltamivir control group( 21. 63 mg/kg per day),Yinhua Pinggan Granule group( 12 g/kg per day),and San-ao Decotion group( 9 g/kg per day). The animals were received gavage administration with corresponding drugs,once per day for 5 successive days. After the gavage administration,lungs,spleens and thymuses were removed and weighed,respectively. Lung index,inhibition ratio of lung index and the immune organ index were calculated. The expressions of viral load in lung tissue of each group were detected via Real-time fluorescent quantitative PCR. The pathological change of lung tissue in each group were detected by hematoxylin-eosin staining( HE) staining. In vitro experiments,Yinhua Pinggan Granule and San-ao Decoction were used to treat Madin-Darby Canine Kidney( MDCK) cells infected with influenza A/H1 N1 virus. The antiviral efficiacy of two formulas against influenza A/H1 N1 virus were detected by MTT method and the cytopathic effects( CPE) that were induced by influenza virus were observed by Microscopy. Results In vivo experiments showed that compared with the normal control group,lung index increased,spleen index and thymus index decreased,multiple lung consolidation,inflammatory cell infiltration,and high expression of viral load in lung tissue of influenza virus model group(P < 0. 01). Compared with influenza virus model group,Yinhua Pinggan Granule and San-ao Decoction significantly decreased lung index,increased spleen index and thymus index in mice infected with influenza virus(P < 0. 05,P < 0. 01),and significantly

关 键 词：银花平感颗粒 三拗汤 甲型H1N1流感病毒 方剂配伍 

分 类 号：R285.5[医药卫生—中药学]