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。
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(20,40,80μL·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.
作者
张美琦
王浩嘉
李艺颖
王靳勇
王雪娇
王遥
贾鑫
ZHANG Meiqi;WANG Haojia;LI Yiying;WANG Jinyong;WANG Xuejiao;WANG Yao;JIA Xin(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;School of Life Science,Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《药物评价研究》
CAS
2022年第9期1697-1705,共9页
Drug Evaluation Research
基金
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)。
