RNAi-Mediated Down-Regulation of CD47 Protects against Ischemia/Reperfusion-Induced Myocardial Damage via Activation of eNOS in a Rat Model
RNAi-Mediated Down-Regulation of CD47 Protects against Ischemia/Reperfusion-Induced Myocardial Damage via Activation of eNOS in a Rat Model
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在大鼠模型中，RNAi 介导的 CD47 下调可通过激活 eNOS 来防止缺血/再灌注引起的心肌损伤
DOI：
10.1159/000453170
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发表时间：
2016-01-01
影响因子：
--
通讯作者：
Yang, Jian
中科院分区：
文献类型：
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作者：
Wang, Hui-bo;Yang, Jun;Yang, Jian
背景/目的：氧化应激与缺血再灌注（I/R）引起的心肌损伤的发病机制密切相关。先前的研究已经证实心脏CD 47驱动左心室心力衰竭。然而，CD 47在心肌I/R损伤（MIRI）中的作用尚未提出。本研究旨在探讨利用RNA干扰（RNAi）技术下调CD 47表达是否能解除一氧化氮（NO）信号通路的抑制，减轻大鼠I/R心肌损伤。研究方法：将雄性Sprague-Dawley大鼠（n = 40）随机分为四组，并用盐水（Sham和I/R组）或表达对照（Ad-EGFP-N）或靶向CD 47（Ad-EGFP-CD 47）RNAi的腺病毒进行预处理。4 d后，结扎大鼠冠状动脉左前降支30 min，再灌注3 h，建立MIRI模型。收集心脏组织，并通过免疫组织化学、蛋白质印迹和定量RT-PCR进行评估。结果指标包括梗死面积、血清心肌酶（肌酸激酶、肌酸激酶MB和乳酸脱氢酶）水平、氧化应激标志物和心肌形态学变化。结果：Ad-EGFP-CD 47 RNAi转染心肌后，心肌组织中CD 47的表达明显降低。CD 47的下调与心肌梗死面积和血清心肌酶水平降低、内皮型一氧化氮合酶活性增加、一氧化氮水平升高和氧化应激水平降低显著相关。结论：这些数据表明，下调CD 47对MIRI发挥保护作用，这可能是由于通过激活eNOS/NO信号通路减轻氧化应激。(C)2016作者（s）由S. Karger AG，巴塞尔
Background/Aims: Oxidative stress is strongly implicated in the pathogenesis of myocardial damage caused by ischemia reperfusion (I/R). Previous studies have confirmed that cardiac CD47 drives left ventricular heart failure. However, the role for CD47 in myocardial I/R injury (MIRI) has not previously been proposed. This study was designed to investigate whether down-regulation of CD47 using RNA interference (RNAi) technology can relieve inhibition of nitric oxide signaling and attenuate myocardial damage in a rat model of I/R. Methods: Male Sprague-Dawley rats (n = 40) were randomly allocated to four groups and pre-treated either with saline (Sham and I/R groups), or adenovirus expressing either control (Ad-EGFP-N) or CD47-targeting (Ad-EGFP-CD47) RNAi. After four days, the rat MIRI model was established by occluding the left anterior descending coronary artery for 30 min, followed by reperfusion for 3 h. Heart tissue was harvested and assessed by immunohistochemistry, western blot, and quantitative RT-PCR. Outcome measures included infarct size, myocardial enzyme (creatine kinase, creatine kinase-MB, and lactate dehydrogenase) levels in serum, markers of oxidative stress, and morphological changes to the myocardium. Results: Delivery of Ad-EGFP-CD47 RNAi into the myocardium remarkably decreased CD47 expression levels. Down-regulation of CD47 was significantly associated with reduced infarct size and serum levels of myocardial enzymes, increased activity of endothelial nitric oxide synthase, increased levels of nitric oxide, and decreased levels of oxidative stress. Conclusion: These data indicate that down-regulation of CD47 exerts a protective effect against MIRI, which may be attributable to attenuation of oxidative stress via activation of the eNOS/NO signaling pathway. (C) 2016 The Author(s) Published by S. Karger AG, Basel