Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine, Daegu 41062, Korea.
Molecules. 2021 Feb 15;26(4):1029. doi: 10.3390/molecules26041029.
Calcium (Ca) dependent signaling circuit plays a critical role in influenza A virus (IAV) infection. The 8--(--methoxycinnamoyl)harpagide (MCH) exhibits pharmacological activities that exert neuroprotective, hepatoprotective, anti-inflammatory and other biological effects. However, not have reports of antiviral effects. To investigate the antiviral activity of MCH on IAV-infected human lung cells mediated by calcium regulation. We examined the inhibitory effect of MCH on IAV infections and measured the level of viral proteins upon MCH treatment using Western blotting. We also performed molecular docking simulation with MCH and IAV M2 protein. Finally, we analyzed MCH's suppression of intracellular calcium and ROS (reactive oxygen species) in IAV-infected human lung cells using a flow cytometer. The results shown that MCH inhibited the infection of IAV and increased the survival of the infected human lung cells. The levels of IAV protein M1, M2, NS1 and PA were inhibited in MCH-treated human lung cells compared to that in infected and untreated cells. Also, docking simulation suggest that MCH interacted with M2 on its hydrophobic wall (L40 and I42) and polar amino acids (D44 and R45), which formed intermolecular contacts and were a crucial part of the channel gate along with W41. Lastly, MCH inhibited IAV infection by reducing intracellular calcium and mitochondrial Ca/ROS levels in infected human lung cells. Taken together, these data suggest that MCH inhibits IAV infection and increases the survival of infected human lung cells by suppressing calcium levels. These results indicate that MCH is useful for developing IAV treatments.
钙（Ca）依赖性信号通路在甲型流感病毒（IAV）感染中发挥着关键作用。8--（-甲氧基肉桂酰基）哈巴苷（MCH）具有药理活性，发挥神经保护、肝保护、抗炎等生物学作用。但是，没有关于其抗病毒作用的报道。为了研究钙调节介导的 MCH 对 IAV 感染人肺细胞的抗病毒活性。我们通过 Western blot 检测了 MCH 对 IAV 感染的抑制作用，并在 MCH 处理后测量了病毒蛋白的水平。我们还进行了 MCH 与 IAV M2 蛋白的分子对接模拟。最后，我们使用流式细胞仪分析了 MCH 在 IAV 感染人肺细胞中对细胞内钙和 ROS（活性氧）的抑制作用。结果表明，MCH 抑制了 IAV 的感染并增加了感染人肺细胞的存活率。与感染和未处理的细胞相比，MCH 处理的人肺细胞中 IAV 蛋白 M1、M2、NS1 和 PA 的水平降低。此外，对接模拟表明 MCH 与 M2 相互作用于其疏水性壁（L40 和 I42）和极性氨基酸（D44 和 R45），形成分子间接触，是与 W41 一起形成通道门的关键部分。最后，MCH 通过降低感染人肺细胞中的细胞内钙和线粒体 Ca/ROS 水平来抑制 IAV 感染。总之，这些数据表明 MCH 通过抑制钙水平抑制 IAV 感染并增加感染人肺细胞的存活率。这些结果表明 MCH 可用于开发 IAV 治疗方法。
