黄芩苷通过促进Ⅰ型干扰素表达抑制鸡传染性支气管炎病毒复制

doi:10.11843/j.issn.0366-6964.2023.11.037

摘要/Abstract

摘要： 本研究旨在探讨黄芩苷对鸡传染性支气管炎病毒(infectious bronchitis virus,IBV)的抑制作用及其作用机制。采用H1299和Vero细胞系作为供试细胞,分别在IBV感染前及感染后在细胞培养体系中加入黄芩苷(125 μg·mL^-1),分析黄芩苷对IBV的抑制作用。进一步,将黄芩苷倍比稀释成不同浓度,观察不同浓度黄芩苷对IBV的抑制作用。将试验分为空白组、IBV组、病毒感染前加药组、病毒感染后加药组,利用实时荧光定量PCR(qRT-PCR)和Western blot技术检测各组干扰素(interferon,IFN)通路相关信号分子的mRNA和蛋白表达情况。用重组人IFN-α处理IBV组和加药组后分析各组IBV对IFN-α治疗的敏感性。以间接免疫荧光技术检测IBV组和加药组磷酸化STAT1(p-STAT1)的核移位。采用基因沉默试验分析IBV组和加药组沉默STAT1对病毒复制的影响。结果显示,黄芩苷对IBV的复制具有一定的抑制作用,并呈剂量依赖性。与对照组相比,黄芩苷处理促进干扰素通路信号分子p-IRF3、IFN-β、p-STAT1的表达水平(P<0.05),下调JAK-STAT信号通路负调控因子SOCS3和SOCS3刺激因子IL-6的表达水平(P<0.05),增加p-STAT1核移位,并增强IBV对IFN-α治疗的敏感性。进一步,沉默STAT1减弱了黄芩苷对IBV的抑制作用。综上所述,黄芩苷可通过诱导Ⅰ型干扰素产生来抑制IBV的复制,减轻病毒感染对宿主细胞造成的细胞病变效应。

关键词: 黄芩苷, 传染性支气管炎病毒(IBV), 复制, 干扰素

Abstract: This study aimed to investigate the inhibitory effect of baicalin on IBV and its underlying mechanism. Firstly, baicalin (125 μg·mL^-1)was added into cells (H1299 and Vero) before or after IBV infection to analyze the inhibitory effect of baicalin on IBV replication, and then baicalin was diluted into different concentrations to observe the inhibitory effect of different concentrations of baicalin on IBV replication. Furthermore, the experiment was divided into normal group, IBV group, pre-treatment group and post-treatment group, qRT-PCR and Western blot were used to detect mRNA and protein expressions of IFN pathway related signaling molecules in each group. At the same time, the sensitivity of IBVs to IFN treatment was analyzed after treatment with recombinant human IFN-α and the nuclear translocation of p-STAT1 was detected by indirect immunofluorescence in IBV group and dosed groups. Finally, the effect of STAT1 silencing on IBV replication was analyzed by gene silencing assay. The results showed that baicalin had an inhibitory effect on IBV replication in a dose-dependent manner. Baicalin treatment up-regulated the expressions of IFN pathway signaling molecules P-IRF3, IFN-β, p-STAT1 (P<0.05), down-regulated the expression of JAK-STAT signaling pathway negative regulator SOCS3 and SOCS3 stimulator IL-6 (P<0.05). Simultaneously, baicalin could increase IFN-mediated translocation of p-STAT1 and increase the sensitivity of IBV to IFN treatment. Further experiments revealed that silencing STAT1 decreased the inhibition effect of baicalin on IBV replication. The results showed that baicalin can inhibit IBV replication and reduce the pathological effect of viral infection on host cells by inducing type Ⅰ IFN production.

Key words: baicalin, infectious bronchitis virus(IBV), replication, interferon

中图分类号:

贾艺泉, 于梦婷, 刘卫容, 方守国, 章松柏. 黄芩苷通过促进Ⅰ型干扰素表达抑制鸡传染性支气管炎病毒复制[J]. 畜牧兽医学报, 2023, 54(11): 4839-4850.

JIA Yiquan, YU Mengting, LIU Weirong, FANG Shouguo, ZHANG Songbai. Baicalin Inhibits Infectious Bronchitis Virus Replication via Promoting the Expression of Type Ⅰ Interferon[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4839-4850.

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