Construction of mTNFR1shRNA Plasmid and its Biological Effects on MHV-3 Induced Fulminant Hepatitis in BALB/cJ Mice  

作　　者：Sui GAO Ming WANG Jian-wen GUO Dong Xi Xiao-ping LUO Qin NING 

机构地区：[1]Laboratory of Infectious Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

出　　处：《Virologica Sinica》2010年第1期52-58,共7页中国病毒学（英文版）

基　　金：National Science Fund of China(NSFC)(30571643,30672380,30700702);National Key Basic Research Program of China(2005CB522901,2007CB512900)

摘　　要：Previous study on TNFRl-mediated hepatocyte apoptosis has been implicated in the development of fulminant viral hepatitis. To interfere with the potentially effective target, plasmid named p-mTNFRlshRNA complimentary to the sequence responsible for mTNFR1 was also constructed and further confirmed by sequence analysis. To investigate the effect of mTNFRlshRNA plasmid on mTNFR1 expression in vivo and the disease progress in MHV-3 induced fulminant hepatitis mice model. By hydrodynamic injection of mTNFRlshRNA plasmid, the survival rate of mice, hepatic pathological change were examined and compared between mice with/without mTNFRlshRNA plasmid intervention. The expression of mTNFR1 was detected by Real-time PCR, immunohistochemistry staining. The mTNFRlshRNA 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.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.

关 键 词：Fulminant hepatitis TNFR1 RNA interference SHRNA 

分 类 号：S858.32[农业科学—临床兽医学] TS218[农业科学—兽医学]