Mechanism by which ma-xing-shi-gan-tang inhibits the entry of influenza virus
Mechanism by which ma-xing-shi-gan-tang inhibits the entry of influenza virus
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DOI：
10.1016/j.jep.2012.05.061
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发表时间：
2012-08-30
影响因子：
5.4
通讯作者：
Horng, Jim-Tong
中科院分区：
文献类型：
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作者：
Hsieh, Chung-Fan;Lo, Cheng-wei;Horng, Jim-Tong
人种药理学相关性：麻杏石甘汤（MXSGT，又名麻杏石甘散）是一种中药汤剂，已被用于治疗普通感冒，发烧和流感病毒感染。然而，其抗流感病毒活性的潜在机制尚未完全了解。在这项研究中，我们研究了MXSGT在流感病毒感染的MDCK细胞的抗病毒作用及其机制，包括病毒表面超微结构的损伤和随之而来的抑制病毒entry.Materials和Methods：无毒浓度的MXSGT对流感病毒A/WSN/33的抗病毒活性进行了检查通过测定（中和试验）其抑制病毒诱导的细胞病变效应。首先用同步感染的添加时间测定法检查MXSGT作用模式，然后进行病毒附着和渗透测定。病毒的内吞作用进行了评价与附着和渗透试验。我们还进行了与抑制病毒进入相关的测定，如神经氨酸酶活性、血凝素活性和磷酸肌醇-3-激酶（PI 3 K）/AKT磷酸化测定。通过定量实时PCR、免疫印迹和免疫荧光显微镜证明了对病毒复制的抑制。结果：MXSGT对A/WSN/33（H1N1）流感病毒的EC 50为0.83 +/- 0.41 mg/ml，具有广谱的抗人流感病毒A/WSN/33（H1N1）的活性，包括对奥司他韦临床耐药株和H1N1 pdm株。当细胞用MXSGT处理时，病毒RNA和蛋白质的合成都被深深地抑制。添加时间试验表明，MXSGT阻断病毒进入期。这通过附着和渗透试验得到证实，其中MXSGT显示出相似的抑制效力（IC 50为0.58 +/- 0.07和0.47 +/- 0.08 mg/ml）。用原子力显微镜进行的高分辨率图像和定量测量证实，病毒表面结构被MXSGT破坏。我们还建立了由PI 3 K/AKT信号通路调节的病毒进入，被MXSGT.Conclusions废除：我们的研究结果为使用MXSGT治疗流感病毒感染提供了科学支持。MXSGT在流感病毒感染的季节性大流行的管理中具有潜在的效用，就像其他临床可用的药物一样。(c)2012爱思唯尔爱尔兰有限公司保留所有权利。
Ethnopharmacological relevance: Ma-xing-shi-gan-tang (MXSGT, aka maxing shigan powder), a Chinese herbal decoction, has been used for the treatment of the common cold, fever, and influenza virus infections. However, the underlying mechanisms of its activity against the influenza virus are not fully understood. In this study, we examined the antiviral effects of MXSGT in influenza-virus-infected MDCK cells and their underlying mechanisms, including the damage of the viral surface ultrastructure and the consequent inhibition of viral entry.Materials and methods: The antiviral activity of nontoxic concentrations of MXSGT against influenza virus A/WSN/33 was examined by assaying (neutralization assay) its inhibition of the virus-induced cytopathic effects. The mode of MXSGT action was first examined with a time-of-addition assay of synchronized infections, followed by viral attachment and penetration assays. Viral endocytosis was evaluated with attachment and penetration assays. We also performed assays related to the inhibition of viral entry, such as neuraminidase activity, hemagglutinin activity, and phosphoinositide-3-kinase (PI3K)/AKT phosphorylation assays. The inhibition of viral replication was demonstrated by quantitative real-time PCR, immunoblotting, and immunofluorescence microscopy. The surface ultrastructure of the MXSGT-treated virus was revealed by atomic force microscopy.Results: MXSGT exhibited an EC50 of 0.83 +/- 0.41 mg/ml against influenza virus A/WSN/33 (H1N1), with broad-spectrum inhibitory activity against different strains of human influenza A viruses, including clinical oseltamivir-resistant isolates and an H1N1 pdm strain. The synthesis of both viral RNA and protein was profoundly inhibited when the cells were treated with MXSGT. The time-of-addition assay demonstrated that MXSGT blocks the virus entry phase. This was confirmed with attachment and penetration assays, in which MXSGT showed similar inhibitory potencies (IC50 of 0.58 +/- 0.07 and 0.47 +/- 0.08 mg/ml). High-resolution images and quantitative measurements made with atomic force microscopy confirmed that the viral surface structure was disrupted by MXSGT. We also established that viral entry, regulated by the PI3K/AKT signaling pathway, was abolished by MXSGT.Conclusions: Our results give scientific support to the use of MXSGT in the treatment of influenza virus infections. MXSGT has potential utility in the management of seasonal pandemics of influenza virus infections, like other clinically available drugs. (c) 2012 Elsevier Ireland Ltd. All rights reserved.