中药组方通过黄酮多酚类抗氧化物质多靶点调节氧化还原状态阻遏病毒感染与炎性反应

Traditional Chinese medicine regulates redox state to suppress viral infection and inflammatory response by the multi-target intervention of flavonoid-polyphenol antioxidants

关键词:  麻黄汤  多靶点抗氧化  呼吸道病毒感染  炎性反应  黄酮多酚类抗氧化物质

Key words:  Mahuang Decoction  multi-target antioxidant  respiratory virus infection  inflammatory response  flavonoid-polyphenol antioxidant

摘要:

[目的] 探究麻黄汤4味中药中黄酮多酚类化合物基于抗氧化抑制新型冠状病毒感染及减轻肺组织炎症损伤的氧化还原机制。[方法] 使用体外自由基医学检测技术评估麻黄汤组方成分的抗氧化特性，包括清除活性氧超氧阴离子（O₂·^-）、羟基自由基（·OH）和氮中心自由基能力以及总抗氧化能力和多酚含量。并使用麻黄汤单味药及其抗氧化单体干预病毒感染的细胞，检测分析细胞内O₂·^-和病毒核心蛋白水平变化。体内实验采用脂多糖（LPS）急性肺部炎症小鼠模型，检测小鼠血清细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平、脾组织巨噬细胞及肺组织氧化还原相关指标核因子红系2相关因子2（NRF2）、硫氧还蛋白1（TRX1）、丙二醛（MDA）和3硝基酪氨酸（3NT）变化。[结果] 结果显示麻黄、桂枝、炙甘草和杏仁4味药中，麻黄具有最强的清除·OH和氮中心自由基能力以及最强的总抗氧化能力，且多酚含量也最高，桂枝具有最强的清除O₂·^-能力，炙甘草和杏仁抗氧化能力则相对较弱。桂枝干预可抑制严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）及其德尔塔（Delta）变体病毒在胞内的复制，并可在体内通过上调抗氧化通路NRF2-TRX1抑制活性氧分子信号从而抑制炎症。此外，组方中含有的黄酮类单体槲皮素也显示对O₂·^-和·OH都有很强的清除能力，并可明显抑制病毒复制。[结论] 研究结果提示，以麻黄和桂枝为代表的含黄酮多酚类抗氧化物质的中药组方成分可以通过不同抗氧化特征发挥多靶点氧化还原调节作用，在病毒入侵、复制和炎性反应的不同氧还状态阶段，发挥不同程度的调控和干预作用，实现对不同病程的有效干预。通过揭示中药组方成分中黄酮多酚类抗氧化物质抗病毒感染和阻遏炎性反应的作用机制，将有助于指导中药更合理配伍，实施精准干预。研究结果也提示，在临床早期干预阶段结合中药抗氧化有效成分的使用，有望为未来可能发生的变异病毒感染的广谱性治疗提供更多的临床获益，从而有助防控新发突发呼吸道病毒的流行，为探索中医药传统辨证施治理论结合抗氧化干预元素以更有效地实现“标本兼治”提供新思路。

Abstract:

[Objective] To investigate the redox mechanism of the flavonoid-polyphenol antioxidants in the four herbs of Mahuang Decoction to inhibit SARS-CoV-2 infection and reduce inflammatory injury of lung tissue. [Methods] The antioxidant properties of the components in Mahuang Decoction were assessed using the free radical medical detection technique，including scavenging abilities of superoxide anion，hydroxyl radical，and DPPH radical，as well as the total antioxidant capacity and total polyphenol content. We used herbs and antioxidant monomers from Mahuang Decoction to intervene in virus-infected cells and assayed to analyze changes in intracellular O₂·^- and the viral core protein level. In vivo experiment was performed using the Lipopolysaccharide（LPS）-induced mouse model. The mice were examined for the levels of tumor necrosis factor-α（TNF-α） and interleukin-6（IL-6），changes of macrophages in spleen and changes of inflammation and redox indexes of nuclear factor erythroid 2-related factor 2（NRF2）， thioredoxin-1（TRX1）， malondialdehyde（MDA），and 3-nitrotyrosine（3NT） in lung tissue. [Results] The results showed that among Mahuang，Guizhi，Zhigancao and Xingren，Mahuang exhibited the strongest scavenging abilities of hydroxyl and DPPH radicals and the strongest total antioxidant capacity，as well as the highest polyphenol content. Guizhi had the strongest scavenging ability of superoxide anion，while the antioxidant capacity of Zhigancao and Xingren was relatively weaker. The intervention of virus-infected cells using Guizhi was found to inhibit the replication of SARS-CoV-2 and Delta variant. Guizhi inhibited ROS signaling and suppressed the inflammatory response by upregulating the NRF2-TRX1 antioxidant pathway. In addition，quercetin，a flavonoid monomer in Mahuang Decoction exhibited strong scavenging abilities of superoxide anion and hydroxyl radicals，and was able to inhibit viral replication obviously. [Conclusion] The flavonoid-polyphenol antioxidants represented by Mahuang and Guizhi could play a multi-targeted role in redox regulation and exert different roles in different stages of viral infection，so as to realize effective and precise intervention. By revealing the mechanism of action of flavonoid-polyphenol antioxidants in antiviral infections and inhibition of inflammatory responses，these results would help to guide the rational compatibility of traditional Chinese medicine ingredients to achieve precise intervention during the treatment of different clinical courses of illnesses. The results of this study also suggested that the antioxidant intervention of traditional Chinese medicine could be used to control respiratory viral infections and prevent new pandemic disasters in a broad-spectrum manner，and provide new ideas for exploring the combination of antioxidant strategies and traditional Chinese medicine.