猪圆环病毒2型诱导PK-15细胞产生IFN-β的信号通路研究

doi:10.3864/j.issn.0578-1752.2016.10.016

Abstract

Abstract: 【Objective】 The objective of this study is to investigate the mechanism of IFN-β production induced by PCV2 in PK-15 cells and provide a theoretical foundation for the pathogenesis and prevention of porcine circovirus disease.【Method】 The PK-15 cells were divided into five groups randomly: Control group, PCV2 group, BX795 (inhibitor of TBK1/IKKε) group, and BAY11-7082 (inhibitor of NF-κB) group, BX795+BAY 11-7082(mixed inhibitors) group. BX795 group, BAY 11-7082 group and BX795+BAY 11-7082 group were pretreated with 0.5µmol BX795, 5µmol BAY 11-7082 and 0.5µmol BX795+5µmol BAY 11-7082 for one hour, respectively. Then the PK-15 cells were infected with PCV2 for 1 h except the control group. The cells were collected at 3, 12, 24, 48 and 72h after infection. The RNA was extracted from the five groups and then reverse-transcribed into cDNA. The mRNA levels of IFN-β, pattern recognition receptors (TLR3, TLR9, RIG-1, MDA-5 and DAI) and adaptor proteins (TRIF, MyD88, Sting, MAVS and IRF3) were detected by real time PCR.【Result】The mRNA levels of IFN-β in PCV2-infected groups were significantly higher than the control at 48h and 72h (P＜0.01), indicating that IFN-β can be induced by PCV2 in PK-15 cells. The mRNA levels of TLR9 in PCV2-infected groups were higher than the control at 48h and 72h (P＜0.01), the mRNA levels of TLR3 increased obviously at 48h (P＜0.01), and the mRNA levels of TRIF and MyD88 which were adaptor proteins of TLR9 and TLR3 were also up-regulated at 48h (P＜0.01), demonstrating that NF-κB signal pathway mediated by TLRs is activated by PCV2. The mRNA levels of MDA-5 and RIG-1 were significantly higher than the control at 72h (P＜0.01), the expression levels of IRF3, MAVS, Sting which were adaptor proteins of RIG-1 and MDA-5 significantly increased at 48h (P＜0.01), indicating that IRF3 signal pathway mediated by RLRs is activated by PCV2. The mRNA levels of DAI were higher at 48h and 72h (P＜0.01), also indicating that DNA recognition receptor is involved in the production of IFN-β. BX795 and BAY 11-7082 inhibitors were used to inhibit the signal pathway mediated by IRF3 and NF-κB, the result showed that the mRNA levels of IFN-β in BX795-treated group was obviously lower than the control group (P＜0.01), while no significant difference was found between BAY 11-7028-treated group and the control group, demonstrating that the up-regulation of IFN-β induced by PCV2 is mainly relative to the IRF3 signal pathway. 【Conclusion】The PCV2 could induce production of IFN-β in PK-15 cells and IRF3 signal pathway played an important role during this process.

Key words: PCV2, PK-15 cell, IFN-β, signal pathway, adaptor proteins

ZHANG Xin-chen, ZHAO Qi-ling, CHEN Meng-meng, SUN Jia-rui, BAO En-dong, ZHANG Shu-xia, Lü Ying-jun. Signal Pathway of Interferon-β Induced by Porcine Circovirus Type 2 in PK-15 Cells[J].Scientia Agricultura Sinica, 2016, 49(10): 2008-2016.

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Signal Pathway of Interferon-β Induced by Porcine Circovirus Type 2 in PK-15 Cells

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