中国中药杂志

2013, v.38(11) 1772-1777

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荆芥挥发油及其主要成分抗流感病毒作用与机制研究
Study on effect and mechanism of volatile oil of Schizonepetae Herba and its essential components against influenza virus

何婷;汤奇;曾南;苟玲;刘金伟;杨靖;于柳;王哲;龚锡平;
HE Ting,TANG Qi,ZENG Nan,GOU Ling,LIU Jin-wei,YANG Jing,YU Liu,WANG Zhe,GONG Xi-ping(Cultivation Base of State and Provincial Key Laboratory for Research,Development and Utilization of Traditional Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

摘要(Abstract):

目的:体内、体外实验观察荆芥挥发油及主要成分薄荷酮、胡薄荷酮的抗甲型流感病毒A/PR/8/34(H1N1)作用及其Toll样受体/干扰素(toll-like receptor/interferon,TLR/IFN)信号通路机制。方法:制备流感病毒性肺炎模型小鼠,以预防和治疗方式给药,测定模型动物肺组织病毒滴度以评价药物体内抗H1N1效应。ELISA法观察药物对模型小鼠血清干扰素-α(interferon-alpha,IFN-α),干扰素-β(interferon-beta,IFN-β),白介素-2(interleukin-2,IL-2),白介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)水平的影响及病毒感染狗肾传代细胞(Madin-Darby canine kidney cell,MDCK)上清液中IFN-β分泌水平的影响。实时荧光定量RT-PCR法观察药物对病毒感染MDCK细胞白介素1受体相关激酶-4(IL-1R-associated kinase-4,IRAK4)与Toll样受体3(toll-like receptor-3,TLR3)mRNA表达水平的影响。结果:体内实验表明,荆芥挥发油0.226 mg.kg-1、薄荷酮0.5 mg.kg-1及胡薄荷酮0.19 mg.kg-1治疗方式给药明显降低感染小鼠肺组织病毒滴度;荆芥挥发油0.226 mg.kg-1明显升高感染小鼠血清IFN-α,IL-2含量;薄荷酮0.5 mg.kg-1显著提高血清IFN-β含量。薄荷酮0.5 mg.kg-1、胡薄荷酮0.19 mg.kg-1明显降低病毒感染小鼠血清IL-6含量。荆芥挥发油0.452,0.226 mg.kg-1及胡薄荷酮0.19 mg.kg-1明显降低模型小鼠血清TNF-α含量。体外实验显示,荆芥挥发油0.1 g.L-1、胡薄荷酮0.1 g.L-1显著升高感染细胞上清液中IFN-β含量,并使细胞IRAK4 mRNA表达水平显著升高;薄荷酮0.25 g.L-1使感染细胞IRAK4 mRNA表达显著增强,但明显降低TLR3 mRNA表达。结论:荆芥挥发油、薄荷酮及胡薄荷酮治疗方式给药能显著降低病毒感染小鼠肺组织病毒滴度,显示出体内抗病毒作用,但预防给药效果不明显。体内抗病毒感染机制与提高感染小鼠干扰素(IFN-α,IFN-β),IL-2水平,抑制IL-6,TNF-α分泌有关;细胞模型结果提示药物通过TLR3和TLR7信号通路影响IFN-β的表达。
Objective: To observe the effect of volatile oil of Schizonepetae Herba(VOSH),and its essential components-menthone and pulegone against anti-influenza virus A/PR/8/34(H1N1) in vivo and in vitro,as well as the signaling mechanism of its toll-like receptor/interferon(TLR/IFN).Method: The lung-adapted PR-8 virus model was prepared in mice.They were administered with preventive and therapeutic drugs,and the hemagglutination titer of model animals was determined to evaluate in vivo effect against H1N1.ELISA test was conducted to observe the effect on IFN-α,IFN-β,IL-2,IL-6 and TNF-α in serum,as well as IFN-β secretion in H1N1 infected MDCK supernatant.Real-time RT-PCR was employed to observe the expression levels of IRAK4 and TLR3 mRNA.Result: The in vivo experiment shows that the hemagglutination titer was significantly decreased when the mice were treated with VOSH(0.266 mg·kg-1),menthone(0.5 mg·kg-1) and pulegone(0.19 mg·kg-1) in therapeutic way;VOSH(0.226 mg·kg-1) had a significant effect on increasing serum levels of IFN-α,IL-2;Methone(0.5 mg·kg-1) had a significant effect on increasing serum levels of IFN-β;Methone(0.5 mg·kg-1) and pulegone(0.19 mg·kg-1) had a significant effect on decreasing serum levels of IL-6;VOSH(0.452,0.226 mg·kg-1) and pulegone(0.19 mg·kg-1) had a significant effect on decreasing serum levels TNF-α.The in vitro experiment showed that the expression levels of IRAK4 mRNA and IFN-β were significantly increased in VOHS(0.1 g·L-1) and pulegone(0.1 g·L-1) groups;and the menthone(0.25 g·L-1) group showed a significant rise in the expression levels of IRAK4 mRNA,but a notable decline in TLR3 mRNA.Conclusion: The administration with VOSH,methone and pulegone in therapeutic way can significantly decrease the hemagglutination titer,which demonstrates the anti-virus effect of the administration in therapeutic way,but no notable efficacy of the administration in preventive way.The in vivo anti-virus mechanism is related to regulation of IFN-α,IFN-β and IL-2.

关键词(KeyWords): 荆芥挥发油;薄荷酮;胡薄荷酮;甲型流感病毒(H1N1);病毒滴度;细胞因子;TLR/IFN信号
volatile oil of Schizonepetae Herba;menthone;pulegone;influenza virus A/PR/8/34(H1N1);hemagglutination titer;cytokines

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(30973923);; 2010年度高层次留学人才回国资助人选项目;; 2010年度留学回国人员科技活动项目择优项目(国家人社部人社厅函[2010]411,412号)

作者(Author): 何婷;汤奇;曾南;苟玲;刘金伟;杨靖;于柳;王哲;龚锡平;
HE Ting,TANG Qi,ZENG Nan,GOU Ling,LIU Jin-wei,YANG Jing,YU Liu,WANG Zhe,GONG Xi-ping(Cultivation Base of State and Provincial Key Laboratory for Research,Development and Utilization of Traditional Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

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