Small Interfering RNA Targeting Heme Oxygenase-1 (HO-1) Reinforces Liver Apoptosis Induced by Ischemia-Reperfusion Injury in Mice: HO-1 Is Necessary for Cytoprotection
Small Interfering RNA Targeting Heme Oxygenase-1 (HO-1) Reinforces Liver Apoptosis Induced by Ischemia-Reperfusion Injury in Mice: HO-1 Is Necessary for Cytoprotection
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DOI：
10.1089/hum.2009.049
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发表时间：
2009-10-01
影响因子：
4.2
通讯作者：
Kupiec-Weglinski, Jerzy W.
中科院分区：
文献类型：
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作者：
Ke, Bibo;Shen, Xiu-Da;Kupiec-Weglinski, Jerzy W.
我们已经证明，血红素氧合酶-1（HO-1）的过表达可以预防导致缺血和再灌注损伤（IRI）的肝脏炎症反应。本研究旨在探讨HO-1在肝IRI中的确切作用和机制，通过采用小干扰RNA（siRNA）有效抑制HO-1表达，在体外和体内。使用部分肝叶热缺血模型，向小鼠注射HO-1 siRNA/非特异性对照siRNA或Ad-HO-1/Ad-beta-gal。HO-1 siRNA治疗组血清谷草转氨酶水平升高，肝水肿明显，肝窦充血/胞质空泡化，肝细胞坏死严重。相比之下，Ad-HO-1预处理的动物仅显示出最小的窦状隙充血，而没有水肿/空泡化或坏死。HO-1 siRNA的施用显著增加了局部中性粒细胞积聚和凋亡细胞的频率。用HO-1 siRNA处理的小鼠的特征在于caspase-3活性增加和HO-1表达减少，而给予Ad-HO-1的小鼠显示caspase-3活性降低和HO-1/Bcl-2/Bcl-xL增加，数据通过使用体外细胞培养系统证实。因此，通过使用siRNA方法，本研究证实HO-1提供针对肝IRI的有效细胞保护并调节肝细胞凋亡。事实上，siRNA提供了一个强大的工具，可以用来研究各种肝脏疾病中的基因功能。
We have shown that overexpression of heme oxygenase-1 (HO-1) prevents the liver inflammation response leading to ischemia and reperfusion injury (IRI). This study was designed to explore the precise function and mechanism of HO-1 cytoprotection in liver IRI by employing a small interfering RNA ( siRNA) that effectively suppresses HO-1 expression both in vitro and in vivo. Using a partial lobar liver warm ischemia model, mice were injected with HO-1 siRNA/nonspecific control siRNA or Ad-HO-1/Ad-beta-gal. Those treated with HO-1 siRNA showed increased serum glutamic-oxaloacetic transaminase levels, significant liver edema, sinusoidal congestion/cytoplasmic vacuolization, and severe hepatocellular necrosis. In contrast, Ad-HO-1-pretreated animals revealed only minimal sinusoidal congestion without edema/vacuolization or necrosis. Administration of HO-1 siRNA significantly increased local neutrophil accumulation and the frequency of apoptotic cells. Mice treated with HO-1 siRNA were characterized by increased caspase-3 activity and reduced HO-1 expression, whereas those given Ad-HO-1 showed decreased caspase-3 activity and increased HO-1/Bcl-2/Bcl-xL, data confirmed by use of an in vitro cell culture system. Thus, by using an siRNA approach this study confirms that HO-1 provides potent cytoprotection against hepatic IRI and regulates liver apoptosis. Indeed, siRNA provides a powerful tool with which to study gene function in a wide range of liver diseases.