宣肺败毒方活性组分对新型冠状病毒刺突蛋白诱导炎症及嗅觉损伤的影响

Effects of the active fraction of Xuanfei Baidu Formula on inflammation and olfactory impairment induced by SARS-CoV-2 spike protein

关键词:  宣肺败毒方  新型冠状病毒  固有免疫  炎症反应  嗅觉损伤

Key words:  Xuanfei Baidu Formula  SARS-CoV-2  innate immunity  inflammatory response  olfactory impairment

摘要:

[目的] 通过对宣肺败毒方活性组分（XFBD-C）的体内外药效研究，探讨其对新型冠状病毒刺突蛋白诱导的炎症反应及嗅觉损伤的影响。[方法] 使用宣肺败毒方活性组分干预新型冠状病毒刺突蛋白（S蛋白）诱导的巨噬细胞炎症模型，通过酶联免疫吸附实验（ELISA）检测白细胞介素-6（IL-6）的分泌；通过实时荧光定量聚合酶链反应（RT- qPCR）检测IL-6、白细胞介素-1β（IL-1β）和单核细胞趋化蛋白-1（MCP-1）的表达。进一步通过对单侧嗅基板注射S蛋白构建斑马鱼幼鱼的嗅觉损伤模型，通过RT-qPCR检测活性组分干预后IL-1β的表达变化；通过喂食行为学实验检测幼鱼的觅食活跃程度；通过荧光成像分析幼鱼头部的嗅神经荧光强度和中性粒细胞聚集情况。[结果] 宣肺败毒方活性组分在细胞模型中有效抑制了S蛋白诱导的IL-6、IL-1β和MCP-1的过度表达，在斑马鱼模型中显著抑制了幼鱼体内IL-1β的表达，提高了S蛋白刺激后幼鱼的觅食活跃程度，并且提升了S蛋白注射区域的嗅神经荧光强度、减弱了中性粒细胞在损伤区域内的聚集。[结论] 宣肺败毒方活性组分可以有效抑制新型冠状病毒刺突蛋白诱导的炎症反应及嗅觉损伤，为辨析宣肺败毒方有效组分及其防治病毒感染诱发的嗅觉损伤研究和应用提供了依据。

Abstract:

[Objective] To evaluate the effects of the active fraction of Xuanfei Baidu Formula on inflammation and olfactory impairment induced by SARS-CoV-2 spike protein using in vivo and in vitro models. [Methods] The active fraction of Xuanfei Baidu Formula was used to intervene the macrophage inflammation model induced by SARS-CoV-2 spike protein（S protein）. The secretion of interleukin-6（IL-6） was measured by enzyme-linked immunosorbent assay（ELISA）. Real-time fluorescence quantitative polymerase chain reaction（RT-qPCR） was used to quantify the expression of IL-6，interleukin-1β（IL-1β），and monocyte chemoattractant protein-1（MCP-1）. Further，an olfactory injury model of zebrafish larvae was constructed by micro-injecting S protein into one side of the olfactory placode. The expression change of IL-1β after treatment of the active fraction was measured by RT-qPCR. Feeding behavior experiment was conducted to test their food perception activity. Fluorescence imaging was used to analyze the fluorescence intensity of olfactory nerves and neutrophil aggregation in the heads of zebrafish. [Results] In vitro，the active fraction of Xuanfei Baidu Formula effectively inhibited the overexpression of IL-6，IL-1β，and MCP-1 induced by S protein. in vivo，it significantly inhibited the expression of IL-1β in zebrafish larvae，improved their locomotion and food perception activity，enhanced fluorescence intensity of olfactory nerves in the injured area，and reduced neutrophils accumulation. [Conclusion] The active fraction of Xuanfei Baidu Formula could effectively inhibit the inflammatory responses and olfactory impairment induced by SARS-CoV-2 spike protein，which provides a basis for research and application of the active components of Xuanfei Baidu Formula，as well as its treatment for virus-induced olfactory impairment.