A novel chimeric ribozyme vector produces potent inhibition of ICAM-1 expression on ischemic vascular endothelium.
A novel chimeric ribozyme vector produces potent inhibition of ICAM-1 expression on ischemic vascular endothelium.
复制标题
一种新型嵌合核酶载体可有效抑制缺血性血管内皮细胞的 ICAM-1 表达。
DOI：
10.1002/jgm.697
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Montgomery,RobertA
中科院分区：
文献类型：
--
作者：
Sonnenday,ChristopherJ;Warren,DanielS;Cooke,SaraK;Dietz,HarryC;Montgomery,RobertA
BackgroundInhibition of intercellular adhesion molecule‐1 (ICAM‐1) expression can ameliorate the inflammation induced by ischemia‐reperfusion injury (IRI) in animal models. However, current strategies to reduce ICAM‐1 expression have been limited by the lack of stability, poor specificity, and the transient nature of synthesized regulatory molecules (antisense/ribozyme).MethodsA chimeric expression vector was generated by fusing a ribozyme targeting sequence against ICAM‐1 to stabilizing stem‐loop structures and nuclear localization signals that are components of endogenous U1 small nuclear RNA. Oligonucleotide scanning was used to predict accessible sites for targeting within the rat ICAM‐1 transcript. Efficacy of the chimeric ribozyme vector was tested by transfection of rat aortic endothelial (RAE) cells (in vitro) and intraportal delivery in a rat hepatic IRI model (in vivo).ResultsTransfection of RAE cells with the chimeric ribozyme vector produced potent and specific inhibition of ICAM‐1 mRNA and protein levels by >65%. This reduction in ICAM‐1 expression was accompanied by a proportional decrease in neutrophil adhesion to RAE cells.In vivointraportal delivery of the chimeric targeting vector to rats sustaining hepatic IRI produced a marked reduction in ICAM‐1 expression on liver endothelium after reperfusion.ConclusionsA chimeric ribozyme vector effectively inhibited ICAM‐1 expression in vascular endothelial cells and in rat liver following IRI, demonstrating a novel gene targeting technique that may be ideally suited to clinical applications aimed at ameliorating IRI. Copyright © 2004 John Wiley & Sons, Ltd.
