[關(guān)鍵詞]
[摘要]
目的 探索隱丹參酮在細胞水平的抗病毒作用,從藥物調(diào)控宿主抗病毒免疫角度,闡釋隱丹參酮抑制病毒復(fù)制的分子機制。方法 通過流式細胞術(shù)、實時熒光定量PCR(qRT-PCR)、免疫印跡實驗(Western blotting)等方法檢測隱丹參酮對水皰性口炎病毒(VSV)、甲型流感病毒(H1N1)、腦心肌炎病毒(EMCV)、Ⅰ型單純皰疹病毒(HSV-1)復(fù)制的影響?;诨蚣患治觯℅SEA),結(jié)合qRT-PCR、流式細胞術(shù),分析隱丹參酮抗病毒的作用機制。在干擾素受體IFNAR1敲除A549細胞(Ifnar1-/-A549)中,檢測I型干擾素(IFN-I)通路對隱丹參酮抗病毒功能的影響。結(jié)果 隱丹參酮顯著抑制VSV、H1N1、EMCV和HSV-1病毒的復(fù)制,在病毒進入前期和進入后階段發(fā)揮抗病毒作用,對病毒吸附過程無影響;通過GSEA分析結(jié)合qRT-PCR驗證,證明隱丹參酮促進IFN-I信號通路的激活;在Ifnar1-/-A549細胞中,隱丹參酮的抗病毒功能受到抑制。結(jié)論 隱丹參酮通過促進IFN-I信號通路激活抑制多種病毒復(fù)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.
[中圖分類號]
R285.5
[基金項目]
中國科協(xié)青年人才托舉工程項目(2020-QNRC1-03);國家自然科學(xué)基金資助項目(31900661,82001663)