[关键词]
[摘要]
目的 在细胞水平评价血必净注射液(简称血必净)对水疱性口炎病毒(VSV)、甲型流感病毒(H1N1)、Ⅰ型单纯疱疹病毒(HSV-1)等多种病毒复制的影响,并探讨其与免疫系统相关的作用机制。方法 通过CCK-8法检测血必净对A549细胞活力的影响;采用流式细胞术及实时荧光定量PCR(qRT-PCR)技术检测血必净对VSV-eGFP、H1N1、HSV-1复制的影响;利用碘化丙啶(PI)染色检测血必净对病毒引起的细胞死亡的影响;qRT-PCR检测血必净对干扰素(IFN)及干扰素刺激基因(ISGs)mRNA水平的影响,以及其对病毒引起的促炎细胞因子基因表达的影响。结果 血必净以剂量相关方式抑制VSV、H1N1及HSV-1等病毒的复制(P<0.001),并显著减少病毒诱发的细胞死亡(P<0.05);血必净能有效降低病毒感染诱导的IL1Β、IL6、TNFΑ基因的表达(P<0.05),并抑制IFIT1、IFIT2等ISGs的表达(P<0.05)。结论 血必净能在体外抑制多种病毒的复制,降低病毒诱导的细胞死亡和炎症反应,其抗病毒的机制可能独立于Ⅰ型IFN信号通路。
[Key word]
[Abstract]
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.
[中图分类号]
R285.5
[基金项目]
中国科协青年人才托举工程(2020-QNRC1-03);国家自然科学基金资助项目(31900661,82001663)