《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (8): 2446-2454.doi: 10.16431/j.cnki.1671-7236.2019.08.031

• 预防兽医 • 上一篇    下一篇

神经肽S及其受体对伪狂犬病病毒感染的体外作用研究

蔡紫峰, 唐梦瑶, 黄辉鹏, 陈吉龙, 祁保民, 杨桂红   

  1. 福建农林大学动物科学学院, 闽台动物病原微生物学实验室, 福州 350002
  • 收稿日期:2019-03-26 出版日期:2019-08-20 发布日期:2019-08-17
  • 通讯作者: 杨桂红 E-mail:877813797@qq.com
  • 作者简介:蔡紫峰(1994-),男,福建泉州人,硕士生,研究方向:基础兽医学,E-mail:347663334@qq.com
  • 基金资助:

    国家自然科学基金(31302050);福建农林大学"金山学者"优秀学术新秀人才项目(2014)

Study on in vitro Efficacy of NPS and NPSR on Pseudorabies Virus Infection

CAI Zifeng, TANG Mengyao, HUANG Huipeng, CHEN Jilong, QI Baomin, YANG Guihong   

  1. Key Laboratory of Fujian-Taiwan Animal Pathogen Biology, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2019-03-26 Online:2019-08-20 Published:2019-08-17

摘要:

为了解神经肽S(NPS)及其受体(NPSR)在伪狂犬病病毒(PRV)感染过程中的作用,本试验采用细胞培养、RT-PCR、实时荧光定量PCR及shRNA技术研究NPS和NPSR在PRV体外感染过程中的表达变化,以及NPS和NPSR对病毒基因和相关细胞因子的变化。RT-PCR和实时荧光定量PCR结果显示,PRV感染培养的小鼠胚胎成纤维细胞系(3T3细胞)后12和18 h,NPS和NPSR mRNA的表达水平极显著上升(P<0.01);通过shRNA技术稳定干扰3T3细胞中NPSR表达后,PRV gE基因的表达极显著下调(P<0.01);添加外源性NPS可显著增强PRV感染的3T3细胞PRV gE基因及细胞因子白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的表达水平(P<0.05);添加NPSR抑制剂后,则极显著抑制细胞中PRV gE基因和细胞因子IL-6和TNF-α的表达水平(P<0.01),该作用效果与NPS的作用效果相反。上述结果表明,降低NPSR的表达能有效抑制PRV在3T3细胞上的复制,说明NPSR是PRV有效感染3T3细胞所必需的重要因子;外源性NPS可通过调节炎症细胞因子的表达而加剧PRV感染诱导的炎症反应。本研究结果为深入探索NPS和NPSR在PRV感染动物体内的调节作用奠定了基础。

关键词: 神经肽S(NPS); NPSR; 伪狂犬病病毒(PRV); 3T3细胞

Abstract:

To explore whether NPS and NPSR were involved in the process of pseudorabies virus (PRV) infection, the expression profiles of NPS and NPSR, virus gene and the related cytokines during PRV infection in vitro were studied by cell culture, RT-PCR, Real-time PCR and shRNA methods.The results of RT-PCR and Real-time PCR showed that the expression of NPS and NPSR in mouse embryonic fibroblasts (3T3 cells) infected with PRV increased extremely significantly 12 and 18 h after PRV infection (P<0.01).After stably interfering with the expression of NPSR in 3T3 cells by shRNA technology, the expression of PRV gE gene decreased extremely significantly (P<0.01).Adding exogenous NPS could significantly enhance the expression of PRV gE gene and cytokines IL-6 and TNF-α in 3T3 cells infected with PRV.However, the expression of PRV gE gene and cytokines IL-6 and TNF-α were extremely significantly inhibited by adding NPSR inhibitors (P<0.01), which was contrary to the effect of NPS.These results indicated that the robust expression of NPS and NPSR could effectively inhibit the replication of PRV in 3T3 cells, which indicated that NPSR was an important factor for the effective infection of PRV in 3T3 cells.Exogenous NPS could aggravate the inflammatory response induced by PRV infection by regulating the expression of inflammatory cytokines.These results provided a basis for further exploring the mechanism of NPS and NPSR involving in PRV infection in vivo.

Key words: neuropeptide S (NPS); NPSR; pseudorabies virus (PRV); 3T3 cell

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