目的 在細(xì)胞水平評價(jià)血必凈注射液（簡稱血必凈）對水皰性口炎病毒（VSV）、甲型流感病毒（H1N1）、Ⅰ型單純皰疹病毒（HSV-1）等多種病毒復(fù)制的影響，并探討其與免疫系統(tǒng)相關(guān)的作用機(jī)制。方法 通過CCK-8法檢測血必凈對A549細(xì)胞活力的影響；采用流式細(xì)胞術(shù)及實(shí)時(shí)熒光定量PCR（qRT-PCR）技術(shù)檢測血必凈對VSV-eGFP、H1N1、HSV-1復(fù)制的影響；利用碘化丙啶（PI）染色檢測血必凈對病毒引起的細(xì)胞死亡的影響；qRT-PCR檢測血必凈對干擾素（IFN）及干擾素刺激基因（ISGs）mRNA水平的影響，以及其對病毒引起的促炎細(xì)胞因子基因表達(dá)的影響。結(jié)果 血必凈以劑量相關(guān)方式抑制VSV、H1N1及HSV-1等病毒的復(fù)制（P＜0.001），并顯著減少病毒誘發(fā)的細(xì)胞死亡（P＜0.05）；血必凈能有效降低病毒感染誘導(dǎo)的IL1Β、IL6、TNFΑ基因的表達(dá)（P＜0.05），并抑制IFIT1、IFIT2等ISGs的表達(dá)（P＜0.05）。結(jié)論 血必凈能在體外抑制多種病毒的復(fù)制，降低病毒誘導(dǎo)的細(xì)胞死亡和炎癥反應(yīng)，其抗病毒的機(jī)制可能獨(dú)立于Ⅰ型IFN信號通路。

Objective To evaluate the treatment effect and underlying mechanism of Xuebijing Injection (XBJ) on the replication of vesicular stomatitis virus (VSV), influenza A virus (H1N1), herpes simplex virus 1 (HSV-1) and other viruses in vitro. Methods CCK8 assay was performed to detect the cell viability of different concentrations of XBJ treated on A549 cells. Flow cytometry and qRT-PCR were conducted to analyze viral replications. The cells were stained with propidium iodide (PI) and calculated by high content imaging to determine cell death. Effect of XBJ on the mRNA levels of IFNB1, interferon-stimulated genes (ISGs) (IFIT1, IFIT2, IFI44, IFIH1, OAS2), and proinflammatory cytokines (IL1Β, IL6, TNFΑ) expression were analyzed by qRT-PCR. Results Low cytotoxic concentration (20, 40, 80 μL·mL^-1) of XBJ were selected for subsequent study. XBJ efficiently suppressed VSV, H1N1and HSV-1 replication in a dosedependent manner (P < 0.001). Meanwhile, compared with the control group, virus-induced cell death and inflammatory response were reduced by XBJ treatment (P < 0.05). In addition, XBJ could effectively reduce the expression of IL1B, IL6 and TNFA genes induced by virus infection (P < 0.05), and inhibit the expression of ISGs such as IFIT1 and IFIT2 (P < 0.05). Conclusion XBJ inhibits the replication of several viruses by reducing virus-induced cell death and inflammatory response in an IFN independent manner.

R285.5

中國科協(xié)青年人才托舉工程（2020-QNRC1-03）；國家自然科學(xué)基金資助項(xiàng)目（31900661，82001663）