目的 探討魚腥草Houttuyniae Herba的體內(nèi)外抗病毒作用及其對(duì)抗病毒天然免疫信號(hào)的激活作用。方法 CCK-8法檢測(cè)魚腥草水提物對(duì)非小細(xì)胞肺癌A549細(xì)胞活力的影響。利用表達(dá)綠色熒光蛋白的水皰性口炎病毒（vesicular stomatitis virus expressing green fluorescent protein，VSV-GFP）感染A549細(xì)胞，流式細(xì)胞術(shù)檢測(cè)魚腥草水提物對(duì)病毒復(fù)制的影響；qRT-PCR檢測(cè)魚腥草水提物對(duì)甲型流感病毒（influenza A virus，H1N1）、腦心肌炎病毒（encephalomyocarditis virus，EMCV）和單純皰疹病毒1型（herpes simplex virus type 1，HSV-1）復(fù)制的影響；利用生物信息學(xué)方法探究魚腥草抗病毒機(jī)制并采用qRT-PCR進(jìn)行驗(yàn)證；在野生型和I型干擾素受體1（interferon-I receptor 1，IFNAR1）敲除的A549細(xì)胞中檢測(cè)魚腥草水提物的抗病毒作用，Western blotting檢測(cè)魚腥草水提物對(duì)I型干擾素（interferon-I，IFN-I）通路下游TANK結(jié)合激酶1（TANK binding kinase 1，TBK1）、信號(hào)轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄活化因子1（signal transducer and activator of transcription 1，STAT1）和STAT2磷酸化水平的影響。構(gòu)建H1N1病毒感染小鼠模型，以奧司他韋為陽性對(duì)照藥，考察魚腥草水提物對(duì)感染小鼠肺組織病毒載量和肺組織病理變化的影響。結(jié)果 在細(xì)胞水平，魚腥草水提物顯著抑制VSV、H1N1、EMCV和HSV-1的復(fù)制（P＜0.05、0.01、0.001），且呈劑量相關(guān)性。在H1N1感染小鼠模型中，魚腥草水提物顯著降低肺組織病毒載量（P＜0.05），并改善肺組織的病理損傷。機(jī)制研究表明，魚腥草水提物在MEF、THP-1和PM細(xì)胞中顯著上調(diào)干擾素刺激基因（interferon stimulated genes，ISGs）表達(dá)（P＜0.05、0.01、0.001），并促進(jìn)TBK1、STAT1和STAT2磷酸化（P＜0.05、0.01、0.001）。結(jié)論 魚腥草通過激活基于IFN-I通路的抗病毒天然免疫發(fā)揮體內(nèi)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

國家自然科學(xué)基金面上項(xiàng)目（82474153）；北京市科技新星計(jì)劃課題（20230484342）；北京市自然科學(xué)基金資助項(xiàng)目（7242239）