解表清里方干预甲型H1N1流行性感冒小鼠的抗病毒作用机制  

Mechanism of Antiviral Action of Jiebiao Qingli Fang in Mice with Influenza A(H1N1)

作　　者：马一川 姚睿祺 明雨 晁燕 张洪春[5] MA Yichuan;YAO Ruiqi;MING Yu;CHAO Yan;ZHANG Hongchun(Beijing University of Chinese Medicine,Beijing 100029,China;Shenzhen Nanshan District Hospital of Traditional Chinese Medicine,Shenzhen 518000,China;Institute of Basic Research in Clinical Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Beijing Oriental Yungjia Pharmaceuticals Co,Ltd,Beijing 100029,China;Clinical Research Center for Respiratory Diseases,China-Japan Friendship Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029 [2]深圳市南山区中医院,深圳518000 [3]中国中医科学院中医临床基础医学研究所,北京100700 [4]北京东方运嘉药业有限公司,北京100029 [5]中日友好医院呼吸疾病临床研究中心,北京100029

出　　处：《世界中医药》2024年第14期2091-2097,共7页World Chinese Medicine

基　　金：国家重点研发计划“中医药现代化”重点专项(2022YFC3500800);中国中医科学院流感防治技术体系(ZZ13-025-06)。

摘　　要：目的:探讨解表清里方对甲型H1N1流行性感冒小鼠的抗病毒作用机制。方法:用随机数字法将36只7周龄雌性BalB/c小鼠随机分为空白组、模型组、西药组及解表清里方高剂量组、中剂量组、低剂量组,每组6只。空白组给予等量生理盐水滴鼻,其余组小鼠给予甲型流感病毒液于左侧鼻孔滴鼻,每只50μL。造模2 h后,西药组给予0.037 5 g/(kg·d)磷酸奥司他韦灌胃;解表清里方高剂量组、中剂量组、低剂量组分别给予6.05 g/(kg·d)、3.02 g/(kg·d)、1.51 g/(kg·d)解表清里方灌胃。空白组及模型组同时给予生理盐水0.2 mL灌胃,均1次/d,连续5 d。对比各组肺指数、胸腺指数和脾脏指数,肺组织病理变化,肺组织中乳动物雷帕霉素靶蛋白(mTOR)表达,肺悬液中病毒基因及磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(AKT)、糖原合酶激酶3β(GSK-3β)mRNA表达,肺组织中PI3K、AKT、GSK-3β蛋白表达。结果:动物实验结果显示,与模型组相比,西药组和中药高剂量组能降低小鼠肺指数(P<0.05),其他组别能降低肺指数、升高胸腺指数和脾脏指数,但差异无统计学意义(P>0.05);解表清里方高、中剂量组能减轻小鼠肺部病变程度,西药组、解表清里方高、中剂量组能降低小鼠肺组织中M、NS2病毒基因复制水平(P<0.01或P<0.05);西药组和解表清里方高剂量组能降低小鼠肺组织中mTOR表达水平;西药组、解表清里方高剂量和中剂量组均能显著下调PI3K、AKT和GSK-3β蛋白和mRNA的表达水平(P<0.01或P<0.05)解表清里方低剂量组在各个方面与模型组相差不显著。结论:解表清里方甲型H1N1流感小鼠有抗病毒作用,其作用机制可能与抑制PI3K/AKT/GSK-3β信号通路中关键蛋白PI3K、AKT、GSK-3β、mTOR表达有关。Objective:To investigate the mechanism of the antiviral action of Jiebiao Qingli Fang in mice with influenza A(H1N1).Methods:Thirty-six 7-week-old female BalB/c mice were randomly divided into the blank group,the model group,the western medicine group,and the high-dose,medium-dose,and low-dose groups of Jiebiao Qingli Fang by the random number method,with six mice in each group.The blank group was given an equal amount of saline nasal drops,and the rest of the mice were given influenza A virus solution in the left nostril at 50 μL.After 2 h of modeling,0.037 5 g/(kg·d) of oseltamivir phosphate was administered by gavage in the western medicine group,and 6.05 g/(kg·d),3.02 g/(kg·d),and 1.51 g/(kg·d) of Jiebiao Qingli Fang were administered to the high-dose group,the medium-dose group,and the low-dose group by gavage,respectively.The blank group and the model group were daily given saline 0.2 mL by gavage at the same time for five consecutive days.Lung index,thymus index,spleen index,pathological changes of lung tissue,expression of mammalian target of rapamycin(mTOR) in lung tissue,mRNA expression of the viral genes and phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),and glycogen synthase kinase 3 beta(GSK-3β) in lung suspension,and protein expression of PI3K,AKT,and GSK-3β in lung tissue were compared between the two groups.Results:The results of animal experiments showed that compared with the model group,the western medicine group and the high-dose group of Jiebiao Qingli Fang were able to reduce the lung index of mice(P<0.05),and the other groups were able to reduce the lung index and increase the thymus index and spleen index,but the differences were not statistically significant(P>0.05).The high-dose and medium-dose groups of Jiebiao Qingli Fang were able to reduce the degree of lung lesions in mice,while the western medicine group and the high-dose and medium-dose groups of Jiebiao Qingli Fang were able to reduce the M and NS2 viral gene replication levels in lung tissue of mice(P<0.01 or P

关 键 词：解表清里方 甲型H1N1流行性感冒 小鼠 抗病毒 雷帕霉素靶蛋白 磷脂酰肌醇3激酶/蛋白激酶B/糖原合酶激酶3β信号通路 细胞自噬 细胞凋亡 

分 类 号：R254[医药卫生—中医内科学]