[关键词]
[摘要]
目的 探索隐丹参酮在细胞水平的抗病毒作用,从药物调控宿主抗病毒免疫角度,阐释隐丹参酮抑制病毒复制的分子机制。方法 通过流式细胞术、实时荧光定量PCR(qRT-PCR)、免疫印迹实验(Western blotting)等方法检测隐丹参酮对水疱性口炎病毒(VSV)、甲型流感病毒(H1N1)、脑心肌炎病毒(EMCV)、Ⅰ型单纯疱疹病毒(HSV-1)复制的影响。基于基因集富集分析(GSEA),结合qRT-PCR、流式细胞术,分析隐丹参酮抗病毒的作用机制。在干扰素受体IFNAR1敲除A549细胞(Ifnar1-/-A549)中,检测I型干扰素(IFN-I)通路对隐丹参酮抗病毒功能的影响。结果 隐丹参酮显著抑制VSV、H1N1、EMCV和HSV-1病毒的复制,在病毒进入前期和进入后阶段发挥抗病毒作用,对病毒吸附过程无影响;通过GSEA分析结合qRT-PCR验证,证明隐丹参酮促进IFN-I信号通路的激活;在Ifnar1-/-A549细胞中,隐丹参酮的抗病毒功能受到抑制。结论 隐丹参酮通过促进IFN-I信号通路激活抑制多种病毒复制。
[Key word]
[Abstract]
Objective To investigate effect of cryptotanshinone on viral replication at the cellular level, and to explain molecular mechanism from perspective of immune regulation. Methods Antiviral effect of cryptotanshinone on the replication of vesicular stomatitis virus (VSV), influenza A virus (H1N1), encephalomyocarditis virus (EMCV) and herpes simplex virus type Ⅰ (HSV-1) were studied by flow cytometry, qRT-PCR and Western blotting. Based on Gene Set Enrichment Analysis (GSEA), combined with qRT-PCR and flow cytometry, the antiviral mechanism of cryptotanshinone was analyzed. The effect of type I interferon (IFN-I) pathway on the antiviral function of cryptotanshinone was detected in IFNAR1 knockout A549 cells (Ifnar1-/-A549). Results Cryptotanshinone significantly inhibited replication of vesicular stomatitis virus (VSV), influenza A virus subtype H1N1 (H1N1) and encephalomyocarditis virus (EMCV), exerted its antiviral effect before and after viral entry, but had no effect on viral attachment step. GSEA analysis combined with qRT-PCR proved that cryptotanshinone promoted the activation of IFN-I signaling pathway. The antiviral function of cryptotanshinone was inhibited in Ifnar1-/-A549 cells. Conclusion Cryptotanshinone inhibits replication of various viruses by activating type I interferon pathway.
[中图分类号]
R285.5
[基金项目]
中国科协青年人才托举工程项目(2020-QNRC1-03);国家自然科学基金资助项目(31900661,82001663)