目的 探讨鱼腥草Houttuyniae Herba的体内外抗病毒作用及其对抗病毒天然免疫信号的激活作用。方法 CCK-8法检测鱼腥草水提物对非小细胞肺癌A549细胞活力的影响。利用表达绿色荧光蛋白的水疱性口炎病毒（vesicular stomatitis virus expressing green fluorescent protein，VSV-GFP）感染A549细胞，流式细胞术检测鱼腥草水提物对病毒复制的影响；qRT-PCR检测鱼腥草水提物对甲型流感病毒（influenza A virus，H1N1）、脑心肌炎病毒（encephalomyocarditis virus，EMCV）和单纯疱疹病毒1型（herpes simplex virus type 1，HSV-1）复制的影响；利用生物信息学方法探究鱼腥草抗病毒机制并采用qRT-PCR进行验证；在野生型和I型干扰素受体1（interferon-I receptor 1，IFNAR1）敲除的A549细胞中检测鱼腥草水提物的抗病毒作用，Western blotting检测鱼腥草水提物对I型干扰素（interferon-I，IFN-I）通路下游TANK结合激酶1（TANK binding kinase 1，TBK1）、信号转导和转录活化因子1（signal transducer and activator of transcription 1，STAT1）和STAT2磷酸化水平的影响。构建H1N1病毒感染小鼠模型，以奥司他韦为阳性对照药，考察鱼腥草水提物对感染小鼠肺组织病毒载量和肺组织病理变化的影响。结果 在细胞水平，鱼腥草水提物显著抑制VSV、H1N1、EMCV和HSV-1的复制（P＜0.05、0.01、0.001），且呈剂量相关性。在H1N1感染小鼠模型中，鱼腥草水提物显著降低肺组织病毒载量（P＜0.05），并改善肺组织的病理损伤。机制研究表明，鱼腥草水提物在MEF、THP-1和PM细胞中显著上调干扰素刺激基因（interferon stimulated genes，ISGs）表达（P＜0.05、0.01、0.001），并促进TBK1、STAT1和STAT2磷酸化（P＜0.05、0.01、0.001）。结论 鱼腥草通过激活基于IFN-I通路的抗病毒天然免疫发挥体内外的抗病毒作用。

Objective To explore the antiviral effect of Yuxingcao (Houttuyniae Herba) in vitro and in vivo, as well as its activation effect on natural immune signals against viruses. Methods CCK-8 method was used to detect the effect of Houttuyniae Herba water extract on viability of A549 cells; A549 cells was infected with vesicular stomatitis virus expressing green fluorescent protein (VSV-eGFP), the effect of Houttuyniae Herba water extract on virus replication was detected by flow cytometry; qRT-PCR was used to detect the effect of Houttuyniae Herba water extract on the replication of influenza A virus (H1N1), encephalomyocarditis virus (EMCV) and herpes simplex virus type 1 (HSV-1); Antiviral mechanism of Houttuyniae Herba was explored by bioinformatics methods and validated through qRT-PCR; The antiviral effect of Houttuyniae Herba water extract was detected in wild-type and type I interferon receptor 1 (IFNAR1) knockout A549 cells, and the effects of Houttuyniae Herba water extract on downstream TANK binding kinase 1 (TBK1), signal transducer and activator of transcription 1 (STAT1) and STAT2 phosphorylation levels in interferon-I (IFN-I) pathway were examined by Western blotting. A mouse model infected with H1N1 virus was constructed and oseltamivir was used as a positive control drug to investigate the effect of Houttuyniae Herba water extract on viral load and pathological changes in lung tissue of infected mice. Results At the cellular level, Houttuyniae Herba water extract significantly inhibited the replication of VSV, H1N1, EMCV and HSV-1 (P < 0.05, 0.01, 0.001), and showed a dose-dependent effect. In the H1N1 infected mouse model, Houttuyniae Herba water extract significantly reduced the viral load in lung tissue (P < 0.05), and improved the pathological damage of lung tissue. Mechanism studies showed that Houttuyniae Herba water extract significantly up-regulated the expressions of interferon stimulated genes (ISGs) in MEF, THP-1 and PM cells (P < 0.05, 0.01, 0.001), and promoted the phosphorylation of TBK1, STAT1 and STAT2 (P < 0.05, 0.01, 0.001). Conclusion Houttuyniae Herba water extract exerts antiviral effects both in vitro and in vivo by activating IFN-I pathway based antiviral innate immunity.

R285.5

国家自然科学基金面上项目（82474153）；北京市科技新星计划课题（20230484342）；北京市自然科学基金资助项目（7242239）