天山堇菜总黄酮抗甲型流感病毒的作用研究  

Study on the Anti-influenza A Virus Effect of Total Flavonoids from Viola tianshanica Maxim

作　　者：王雪 史玉柱 司丽君 马雪萍 王林林 姚华 黄华 WANG Xue;SHI Yuzhu;SI Lijun;MA Xueping;WANG Linlin;YAO Hua;HUANG Hua(Xinjiang Institute of Materia Medica,Urumqi 830004 Xinjiang,China)

机构地区：[1]新疆维吾尔自治区药物研究所,新疆乌鲁木齐830004

出　　处：《中药新药与临床药理》2020年第6期655-661,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：新疆维吾尔自治区自然科学基金面上项目(2017D01A71);新疆地产中药民族药新药研发培育项目(2016-02-03)。

摘　　要：目的考察天山堇菜总黄酮提取物(TSMF)体内外抗甲型流感病毒的作用。方法①体外实验:采用细胞病变效应(CPE)评分法,考察天山堇菜总黄酮对流感病毒株A/H1N1/京防/262/95、A/H3N2/济防/15/90、A/H3N2/粤防/243/72感染犬肾细胞(MDCK)引起细胞病变的影响。②体内实验:采用甲型流感病毒鼠肺适应株A/H1N1/PR/8/34感染的小鼠模型,观察天山堇菜总黄酮对流感病毒感染小鼠的生命延长率、死亡保护率、肺指数和肺匀浆中细胞因子干扰素-γ(IFN-γ)、白介素-1β(IL-1β)、白介素-10(IL-10)的含量,以及对血清中肿瘤坏死因子-α(TNF-α)、IFN-γ、IL-10含量的影响,以HE染色法观察天山堇菜总黄酮对流感病毒感染小鼠肺组织病理变化的影响。结果①体外实验中,与病毒对照组比较,天山堇菜总黄酮对流感病毒株A/H1N1/京防/262/95、A/H3N2/济防/15/90、A/H3N2/粤防/243/72引起的MDCK细胞病变均有一定的抑制作用,其选择指数(SI)值分别为:4.7、9.5、16.8。②体内实验中,与模型组比较,天山堇菜总黄酮中、高剂量组的小鼠死亡率显著降低(P <0.05),存活时间明显延长(P <0.01),其生命延长率分别为48.6%、51.2%;中、高剂量组小鼠的肺指数明显降低(P <0.05,P <0.01),抑制率分别为20.8%、27.6%;天山堇菜总黄酮各剂量组肺匀浆中的IL-1β及血清中的TNF-α水平明显降低(P <0.05,P <0.01),肺匀浆中IL-10水平明显升高(P <0.01);天山堇菜总黄酮中、高剂量组肺匀浆及血清中的IFN-γ水平均明显降低(P <0.05,P <0.01),高剂量组的血清IL-10水平明显升高(P <0.05)。此外,天山堇菜总黄酮对流感病毒感染小鼠的肺组织病理损伤具有明显改善作用。结论天山堇菜总黄酮具有较好的抗流感病毒作用,还能够调控炎性细胞因子的分泌,从而减轻流感病毒感染小鼠的肺部炎症。Objective To investigate the anti-influenza A virus effect of total flavonoids extract from Viola tianshanica Maxim.(TSMF)in vitro and in vivo. Methods For in vitro investigation,cytopathic effect(CPE)lesion scoring was adopted to observe the effect of TSMF on influenza virus strains A/H1N1/JINGFANG/262/95,A/H3N2/JIFANG/15/90, and A/H3N2/YUEFANG/243/72 in MDCK cells respectively. For in vivo investigation, ICR mice were infected with influenza virus A/H1N1/PR/8/34, then the life extension rate, death protection rate and lung index were observed;levels of interferon-γ(IFN-γ),interleukin-1β(IL-1β)and interleukin-10(IL-10)in lung homogenates and the levels of tumor necrosis factor(TNF-α),IFN-γ and IL-10 in serum were analyzed. Lung histopathological changes were visualized by HE staining. Results In vitro experiment:compared with virus control group,TSMF had good inhibitory effect on influenza virus strains A/H1N1/JINGFANG/262/95,A/H3N2/JIFANG/15/90 and A/H3N2/YUEFANG/243/72, with the SI values of 4.7, 9.5, and 16.8 respectively. In vivo experiment:compared with the model group,at medium and high doses TSMF could significantly decrease death rate(P < 0.05)and prolong the survival time of the mice infected with influenza virus(P < 0.01),the life extension rates were 48.6% and 51.2% respectively. At medium and high doses,TSMF could significantly decrease the lung index(P <0.05, P < 0.01), the inhibition rates were 20.8% and 27.6%, respectively. Compared with the model group,TSMF could obviously decrease the levels of IL-1β in lung homogenates and TNF-α in serum(P < 0.05,P < 0.01)and increase the levels of IL-10 in lung homogenates(P < 0.01). At medium and high doses, TSMF could significantly decrease the levels of IFN-γ(P < 0.05,P < 0.01)in lung homogenates and in serum. At high dose,TSMF could also obviously increase the levels of IL-10 in lung homogenates and in serum(P < 0.05). Additionally,TSMF could improve lung pathology. Conclusion The total flavonoids from Viola tianshanica Maxim. has a good effect ag

关 键 词：天山堇菜总黄酮 甲型流感病毒 细胞病变效应 肺部炎症 炎性细胞因子 小鼠 

分 类 号：R285.5[医药卫生—中药学]