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Volume 25 Issue 1
February 2010
    Citation: Sui GAO, Ming WANG, Jian-wen GUO, Dong Xi, Xiao-ping LUO, Qin NING. Construction of mTNFR1shRNA Plasmid and its Biological Effects on MHV-3 Induced Fulminant Hepatitis in BALB/cJ Mice .VIROLOGICA SINICA, 2010, 25(1) : 52-58.  http://dx.doi.org/10.1007/s12250-010-3072-x
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    Construction of mTNFR1shRNA Plasmid and its Biological Effects on MHV-3 Induced Fulminant Hepatitis in BALB/cJ Mice

    cstr: 32224.14.s12250-010-3072-x

    • Laboratory of Infectious Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

    • Corresponding author: Qin NING, qning@tjh.tjmu.edu.cn
    • Received Date: 11 May 2009
      Accepted Date: 12 October 2009
      Available online: 01 February 2010

      Fund Project: National Key Basic Research Program of China 2007CB512900National Science Fund of China (NSFC) 30672380National Science Fund of China (NSFC) 30571643National Key Basic Research Program of China 2005CB522901National Science Fund of China (NSFC) 30700702

    • Previous study on TNFR1-mediated hepatocyte apoptosis has been implicated in the development of fulminant viral hepatitis. To interfere with the potentially effective target, plasmid named p-mTNFR1shRNA complimentary to the sequence responsible for mTNFR1 was also constructed and further confirmed by sequence analysis. To investigate the effect of mTNFR1shRNA plasmid on mTNFR1 expression in vivo and the disease progress in MHV-3 induced fulminant hepatitis mice model. By hydrodynamic injection of mTNFR1shRNA plasmid, the survival rate of mice, hepatic pathological change were examined and compared between mice with/without mTNFR1shRNA plasmid intervention. The expression of mTNFR1 was detected by Real-time PCR, immunohistochemistry staining. The mTNFR1shRNA plasmid significantly reduced mTNFR1 expression in vivo, markedly ameliorates inflammatory infiltration, prolonged the survival time period and elevated the survival rate from 0 up to 13.3% in Balb/cJ mice with MHV-3 induced fulminant hepatitis. This study was designed to explore the opportunity of RNA interference technique in inhibiting TNFR1 expression, which has been reported to be involved in the development of a variety of diseases including fulminant viral hepatitis and severe chronic hepatitis B.
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      4. Cheng V C, Lo C M, Lau G K, et al. 2003. Current issues and treatment of fulminant hepatic failure including transplantation in Hong Kong and the Far East. Semin. Liver Dis, 23, 239-250.
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      5. Costelli P, Aoki P, Zingaro B, et al. 2003. Mice lacking TNFalpha receptors 1 and 2 are resistant to death and fulminant liver injury induced by agonistic anti-Fas antibody. Cell Death Differ, 10 (9): 997-1004.
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      6. Elbashir S M, Harborth J, Lendeckel W, et al. 2001. Duplexes of 21-nucleotide RNAs mediate RNA in-terference in cultured mammalian cells. Nature, 411 494-498.
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      Construction of mTNFR1shRNA Plasmid and its Biological Effects on MHV-3 Induced Fulminant Hepatitis in BALB/cJ Mice

        Corresponding author: Qin NING, qning@tjh.tjmu.edu.cn
      • Laboratory of Infectious Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
      • Received Date: 11 May 2009
      • Accepted Date: 12 October 2009
      Fund Project:  National Key Basic Research Program of China 2007CB512900National Science Fund of China (NSFC) 30672380National Science Fund of China (NSFC) 30571643National Key Basic Research Program of China 2005CB522901National Science Fund of China (NSFC) 30700702

      Abstract: Previous study on TNFR1-mediated hepatocyte apoptosis has been implicated in the development of fulminant viral hepatitis. To interfere with the potentially effective target, plasmid named p-mTNFR1shRNA complimentary to the sequence responsible for mTNFR1 was also constructed and further confirmed by sequence analysis. To investigate the effect of mTNFR1shRNA plasmid on mTNFR1 expression in vivo and the disease progress in MHV-3 induced fulminant hepatitis mice model. By hydrodynamic injection of mTNFR1shRNA plasmid, the survival rate of mice, hepatic pathological change were examined and compared between mice with/without mTNFR1shRNA plasmid intervention. The expression of mTNFR1 was detected by Real-time PCR, immunohistochemistry staining. The mTNFR1shRNA plasmid significantly reduced mTNFR1 expression in vivo, markedly ameliorates inflammatory infiltration, prolonged the survival time period and elevated the survival rate from 0 up to 13.3% in Balb/cJ mice with MHV-3 induced fulminant hepatitis. This study was designed to explore the opportunity of RNA interference technique in inhibiting TNFR1 expression, which has been reported to be involved in the development of a variety of diseases including fulminant viral hepatitis and severe chronic hepatitis B.

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