基于唾液酸化的电压依赖性Ca2+通道探讨二氢吡啶类钙通道阻滞剂抗甲型流感病毒的机制研究

流感病毒不断威胁着人类的健康，但传统的抗流感药物均已经出现耐药，迫切需要研发新作用机制抗病毒药物。最新的研究显示，唾液酸化的电压依赖性Ca2+通道Cav1.2是甲型流感病毒感染的关键受体分子，这一发现不仅对流感病毒发病机制提出了全新认识，且为抗病毒药物的研发指出了新方向。我们前期研究发现尼索地平等二氢吡啶类钙通道阻滞剂具有抗甲型流感病毒的活性，但其抗病毒作用机制尚未阐明。本项目采用FRET、GST pull-down、激光共聚焦显微镜技术以及膜片钳等方法，从细胞、分子水平阐明二氢吡啶类钙通道阻滞剂抗流感病毒作用机制，以及评价其与神经氨酸酶抑制剂联用的药效。同时，探索其他RNA病毒如登革病毒内化机制的共性和规律。本项目旨在深入挖掘老药潜力，为流感提供新的防治策略。同时为解析RNA病毒其它成员的内化机制提供新的视角和重要依据，为研发相对广谱的抗病毒入侵药物提供思路。

Influenza virus is constantly threatening human health worldwide. Unfortunately, resistant strains to existing anti-influenza drugs have also emerged. It is urgent to develop novel antiviral drugs with new mechanism of actions for the prevention and treatment of influenza. The latest research shows that a sialylated voltage-dependent Ca2+ Channel is critical for influenza A virus infection.The result not only puts forward a new understanding of the pathogenesis of influenza virus, but also provides a potential target for the development of novel anti-influenza drugs. Our previous studies suggested that dihydropyridine calcium blockers such as nisoldipine exerted a significant inhibitory effect on influenza A virus. In the present study, we attempt to elucidate the mechanism of dihydropyridine calcium blockers against influenza A virus by FRET, GST pull-down, co-localization under laser confocal microscope assay and patch clamp technique, and then evaluate the efficacy of dihydropyridine calcium blockers in combination with neuraminidase inhibitors. To explore the common characteristics and rules of virus entry, the internalization mechanism of other RNA viruses such as dengue viruses will be further studied. The purpose of this project is to enlarge the clinical application of dihydropyridine calcium blockers and provide a new strategy for influenza prevention and control. It also provides a new perspective and an important basis for the analysis of the internalization mechanism of other RNA viruses, which offers a new way for the research and development of broad-spectrum antiviral drugs.