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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2008, Vol. 2 Issue (2) : 166-170     DOI: 10.1007/s11684-008-0030-1
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Blockage of receptor-interacting protein 2 expression by small interfering RNA in murine macrophages
LIU Hongchun, CAO Zhongwei, JIN Jianjun, WANG Jiyao
Department of Gastroenterology, Zhongshan Hospital, Fudan University
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Abstract  This study aims to demonstrate that blocking the receptor-interacting protein2 (Rip2) expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality. Murine Rip2 small interfering RNA (siRNA) plasmids were constructed and transfected into macrophage and Rip2 expression was detected with reverse transcription-polymerase chain reaction (RT-PCR) and western blot. Cell proliferation was assayed with MTT. TNF-? concentration was assayed with ELISA and high-mobility group box 1 protein (HMGB1) level with semi-quantitative western blot after lipopolysaccharide (LPS) stimulation. LPS challenge was given after the plasmids were injected into mice and the survival rate was calculated. Rip2 siRNA plasmid could block the mRNA and protein expression of Rip2 and promote cell proliferation. Blocking Rip2 could attenuate LPS-induced TNF-? and HMGB1 production. The HMGB1 expression in the liver decreased to (40.21 ± 11.03) pg/g, and serum TNF-? level decreased to (300.43 ± 59.26) ng/L (P < 0.05). The survival rate of mice from endotoxemia was also improved (P < 0.05). The results demonstrate that Rip2 siRNA plasmid can block the expression of Rip2, decrease the production of TNF-? and HMGB1 and protect mice from fatal endotoxemia.
Issue Date: 05 June 2008
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-008-0030-1     OR     http://academic.hep.com.cn/fmd/EN/Y2008/V2/I2/166
1 Yang H Ochani M Li J Qiang X Tanovic M Harris H E Susarla S M Ulloa L Wang H DiRaimo R Czura C J Wang H Roth J Warren H S Fink M P Fenton M J Andersson U Tracey K J Reversing established sepsis with antagonists of endogenoushigh-mobility group box 1Proc Natl AcadSci USA 2004 101(1)296301.
doi:10.1073/pnas.2434651100
2 Kobayashi K Inohara N Hernandez L D Galán J E Núñez G Janeway C A Medzhitov R Flavell R A RICK/Rip2/CARDIAK mediatessignaling for receptors of the innate and adaptive immune systemsNature 2002 416(6877)194199.
doi:10.1038/416194a
3 Elbashir S M Lendeckel W Tuschl T RNA interference is mediated by 21- and 22-nucleotide RNAsGenes Dev 2001 15(2)188200.
doi:10.1101/gad.862301
4 Schwarz D S Hutvágner G Du T Xu Z Aronin N Zamore P D Asymmetryin the assembly of the RNAi enzyme complexCell 2003 115(2)199208.
doi:10.1016/S0092‐8674(03)00759‐1
5 McCully M Baroja M Chau T Salgado A Blake P Madrenas J Su.80.up-regulation of Rip2 expression during peritonitis correlates withclinical outcome. Clin Immunol 2006 119(Suppl 1)s187.
doi: 10.1016/j.clim.2006.04.507
6 Nicholson D W Caspasestructure, proteolytic substrates, and function during apoptotic celldeath. Cell Death Differ. 1999 6(11)10281042
7 McCarthy J V Ni J Dixit V M RIP2 is a novel NF-kappaB-activating and cell death-inducingkinaseJ Biol Chem 1998 273(27)1696816975.
doi:10.1074/jbc.273.27.16968
8 Wang H Yang H Tracey K J Extracellular role of HMGB1 in inflammation and sepsisJ Intern Med 2004 255(3)320331.
doi:10.1111/j.1365‐2796.2003.01302.x
9 Jungas T Verbeke P Darville T Ojcius DM Cell death,BAX activation, and HMGB1 release during infection with ChlamydiaMicrobes Infect 2004 6(13)11451155.
doi:10.1016/j.micinf.2004.07.004
10 Wang H Bloom O Zhang M Vishnubhakat J M Ombrellino M Che J Frazier A Yang H Ivanova S Borovikova L Manogue K R Faist E Abraham E Andersson J Andersson U Molina P E Abumrad N N Sama A Tracey K J HMG-1as a late mediator of endotoxin lethality in miceScience 1999 285(5425)248251.
doi:10.1126/science.285.5425.248
11 Andersson U Wang H Palmblad K Aveberger A C Bloom O Erlandsson-Harris H Janson A Kokkola R Zhang M Yang H Tracey K J HMG-1 stimulates proinflammatory cytokinesynthesis in human monocytesJ Exp Med 2000 192(4)565576.
doi:10.1084/jem.192.4.565
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