中国人兽共患病学报2024,Vol.40Issue(5) :421-429.DOI:10.3969/j.issn.1002-2694.2024.00.063

猪源SIRT5促进O型口蹄疫病毒在PK-15细胞复制

Porcine SIRT5 promotes replication of foot and mouth disease virus type O in PK-15 cells

陈国辉 史喜绢 别鑫恬 杨行 赵思越 张大俊 赵登率 闫文倩 陈玲玲 赵美玉 何路 郑海学 刘霞 张克山
中国人兽共患病学报2024,Vol.40Issue(5) :421-429.DOI:10.3969/j.issn.1002-2694.2024.00.063

猪源SIRT5促进O型口蹄疫病毒在PK-15细胞复制

Porcine SIRT5 promotes replication of foot and mouth disease virus type O in PK-15 cells

陈国辉 1史喜绢 2别鑫恬 2杨行 2赵思越 1张大俊 2赵登率 2闫文倩 2陈玲玲 2赵美玉 1何路 2郑海学 2刘霞 3张克山2
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作者信息

  • 1. 甘肃农业大学生命科学技术学院,兰州 730070;中国农业科学院兰州兽医研究所,兰州大学动物医学与生物安全学院动物疫病防控国家重点实验室,兰州 730046
  • 2. 中国农业科学院兰州兽医研究所,兰州大学动物医学与生物安全学院动物疫病防控国家重点实验室,兰州 730046
  • 3. 甘肃农业大学生命科学技术学院,兰州 730070
  • 折叠

摘要

目的 探究猪源SIRT5对O型口蹄疫病毒(Foot and mouth disease virus serotype O,FMDV-O)复制的影响及其调控FMDV-O复制的初步机制.方法 利用Western Blotting和RT-qPCR检测FMDV-O感染PK-15细胞后内源性SIRT5表达情况;设计合成3对SIRT5特异性siRNA,通过Western Blotting和RT-qPCR检测SIRT5、FMDV-O蛋白水平、转录水平及病毒拷贝数的变化情况;构建SIRT5真核表达质粒转染至PK-15细胞,运用Western Blotting、RT-qPCR实验方法探究过表达SIRT5对FMDV-O复制的影响,同时利用RT-qPCR检测过表达SIRT5对SeV、FMDV-O诱导的I型干扰素刺激基因mRNA表达水平的影响.结果 FMDV-O感染PK-15细胞后内源性SIRT5的表达上调;siRNA干扰SIRT5抑制FMDV-O的复制;过表达SIRT5促进FMDV-O复制;过表达SIRT5降低了 SeV、FMDV-O诱导的干扰素刺激基因mRNA的表达水平.结论 FMDV-O感染刺激宿主SIRT5表达,而猪源SIRT5通过抑制I型干扰素刺激基因的产生进而促进FMDV-O复制.本研究为进一步探究猪源SIRT5促进FMDV-O复制的机制提供参考依据.

Abstract

The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O)and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endoge-nous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 over-expression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O rep-lication by inhibiting production of type I interferon-stimula-ted genes.These findings provide a reference to further ex-plore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

关键词

猪源SIRT5/FMDV-O/干扰素刺激基因/PK-15细胞

Key words

porcine SIRT5/FMDV-O/interferon stimulating gene/PK-15 cells

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基金项目

国家重点研发计划项目(2021YFD1800300)

甘肃省科技重大专项(21ZD3NA001-5)

中国农业科学院重大任务(CAAS-ZDRW202006-03)

出版年

2024
中国人兽共患病学报
中国微生物学会

中国人兽共患病学报

CSTPCD北大核心
影响因子:0.814
ISSN:1002-2694
参考文献量2
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