戊型肝炎病毒感染调控TLR3信号通路的研究
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摘要
戊型肝炎病毒(Hepatitis E virus,HEV)是导致急性戊肝的主要原因.通过建立基因IV型HEV感染A549细胞模型,研究病毒感染后TLR3信号通路相关因子表达的变化.病毒感染A549细胞后通过实时荧光定量PCR检测IFN-β的mRNA表达量,Western blot分析磷酸化IRF3(pIRF3)和IKKε蛋白表达水平的变化.结果表明:HEV感染A549细胞后细胞内IFN-β的相对表达水平显著下降,TLR3信号通路介导的pIRF3及IKKε蛋白在病毒感染后表达量显著下降.基因Ⅳ型HEV能够抑制TLR3信号通路介导的IRF3的磷酸化及IKKε蛋白的表达,从而抑制了IFN-β的表达,为进一步研究HEV复制机制和致病机理奠定基础.
Hepatitis E virus(HEV) infection is the leading cause of acute hepatitis E. A549 cell model was established by infected HEV, and the expression of TLR3 signaling pathway-related factors was studied after viral infection. The mRNA expression of IFN-β was detected by real-time quantitative PCR after A549 cells were infected with virus. The expression levels of phosphorylated IRF3(pIRF3) and IKKε protein were analyzed by Western blot. The results showed that the relative expression level of IFN-β was significantly decreased in HEV-infected A549 cells, and the expression of pIRF3 and IKKε protein mediated by TLR3 signaling pathway was significantly decreased after viral infection. Gene type IV HEV can inhibit the expression of phosphorylated IRF3 and IKKε protein mediated by TLR3 signaling pathway, thereby inhibiting the expression of IFN-β, which will facilitate further studies on HEV replication mechanism and pathogenesis.
Hepatitis E virus(HEV) infection is the leading cause of acute hepatitis E. A549 cell model was established by infected HEV, and the expression of TLR3 signaling pathway-related factors was studied after viral infection. The mRNA expression of IFN-β was detected by real-time quantitative PCR after A549 cells were infected with virus. The expression levels of phosphorylated IRF3(pIRF3) and IKKε protein were analyzed by Western blot. The results showed that the relative expression level of IFN-β was significantly decreased in HEV-infected A549 cells, and the expression of pIRF3 and IKKε protein mediated by TLR3 signaling pathway was significantly decreased after viral infection. Gene type IV HEV can inhibit the expression of phosphorylated IRF3 and IKKε protein mediated by TLR3 signaling pathway, thereby inhibiting the expression of IFN-β, which will facilitate further studies on HEV replication mechanism and pathogenesis.
引文
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[3] Aggarwal R,Jameel S.Hepatitise[J].Hepatology,2011,54(6):2218-2226.
[4] Kumagai Y,Akira S.Identification and functions of pattern-recognition receptors[J].Journal of Allergy and Clinical Immunology,2010,125(5):985-992.
[5] Fang J,Fang D,Silver P B,et al.The role of TLR2,TRL3,TRL4,and TRL9 signaling in the pathogenesis of autoimmune disease in a retinal autoimmunity model[J].Investigative ophthalmology & visual science,2010,51(6):3092-3099.
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[8] He M,Wang M,Huang Y,et al.The ORF3 protein of genotype 1 hepatitis E virus suppresses TLR3-induced NF-κB signaling via TRADD and RIP1[J].Scientific reports,2016,6:27597.
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[11] Mogensen T H.Pathogen recognition and inflammatory signaling in innate immune defenses[J].Clinical microbiology reviews,2009,22(2):240-273.
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[13] Jiang L,Wan Y,Anderson J C,et al.Genetic dissection of Arabidopsis MAP kinase phosphatase 1-dependent PAMP-induced transcriptional responses[J].Journal of experimental botany,2017,68(18):5207-5220.
[14] Liu L,Botos I,Wang Y,et al.Structural basis of toll-like receptor 3 signaling with double-stranded RNA[J].Science,2008,320(5874):379-381.
[15] Devhare P B,Chatterjee S N,Arankalle V A,et al.Analysis of antiviral response in human epithelial cells infected with hepatitis E virus[J].PloS one,2013,8(5):e63793.
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[17] Vercammen E,Staal J,Beyaert R.Sensing of viral infection and activation of innate immunity by toll-like receptor 3[J].Clinical microbiology reviews,2008,21(1):13-25.
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