150-kDa oxygen-regulated protein attenuates myocardial ischemia-reperfusion injury in rat heart
150-kDa oxygen-regulated protein attenuates myocardial ischemia-reperfusion injury in rat heart
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DOI：
10.1016/j.yjmcc.2005.01.001
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发表时间：
2005-03-01
影响因子：
5
通讯作者：
Matsuda, H
中科院分区：
文献类型：
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作者：
Aleshin, AN;Sawa, Y;Matsuda, H
早期收缩功能障碍和后期心肌细胞死亡是心肌梗死或需要心脏缺血停搏的重大心血管手术后可能出现的两个主要问题。我们发现24小时缺氧和1小时复氧可诱导培养大鼠心肌细胞中伴侣蛋白ORP150的表达。用腺病毒表达反义ORP150抑制其诱导可显著增强缺氧再氧诱导的心肌细胞死亡；过表达ORP150可减少细胞死亡。ORP150表达水平的降低也增强了caspase-3和-8的激活、细胞色素c的释放和DNA的断裂，表明这种伴侣蛋白调节凋亡细胞的死亡。相反，增加心肌细胞中ORP150的表达对这些分子的表达有相反的影响。此外，用日本血凝病毒(HVJ)-脂质体法将ORP150表达质粒转染到大鼠全心，可在体内显著抑制心肌缺血再灌注（I/R）引起的凋亡细胞死亡。有趣的是，ORP150似乎在体外经历缺血再氧化的心肌细胞中保持钙稳态。反义ORP150增强心肌细胞钙蛋白酶活性，而义ORP150抑制心肌细胞钙蛋白酶活性。最后，我们检测了过度表达ORP150或GFP蛋白并进行I/R的大鼠心脏的功能恢复情况；我们发现ORP150在短暂缺血后保留了早期收缩功能。我们的研究结果表明ORP150在大鼠心脏中具有细胞保护作用，并提示该蛋白在防止心肌细胞死亡和保持缺血损伤后的收缩功能方面具有治疗作用。(c) 2005 Elsevier Ltd版权所有。
Early contractile dysfunction and the later death of cardiomyocytes are two major problems that can follow myocardial infarction or major cardiovascular surgery that demands ischemic arrest of the heart. Here, we found that 24 h of hypoxia and 1 h of reoxygenation induced the expression of the chaperone ORP150 in cultured rat cardiomyocytes. Inhibition of its induction using an adenovirus to express anti-sense ORP150 significantly enhanced the hypoxia-reoxygenation-induced cardiomyocyte death; cell death was reduced by overexpressing ORP150. Decreased levels of ORP150 expression also enhanced caspase-3 and -8 activation, cytochrome-c release, and DNA fragmentation, suggesting that this chaperone regulates apoptotic cell death. In contrast, increasing the expression of ORP150 in the cardiomyocytes had the opposite effect on the expression of these molecules. Moreover, apoptotic cell death initiated by myocardial ischemia-reperfusion (I/R) was significantly inhibited in vivo by transfecting an ORP150 expression plasmid into whole rat heart using the hemagglutinating virus of Japan (HVJ)-liposome method. Interestingly, ORP150 seemed to preserve calcium homeostasis in cardiomyocytes that underwent ischemia-reoxygenation in vitro. Calpain activity in the cardiomyocytes was enhanced by anti-sense ORP150 and suppressed by sense ORP150. Finally, we examined the functional recovery of rat hearts that overexpressed ORP150 or GFP protein and were subjected to I/R; we found that ORP150 preserved early contractile function after transient ischemia. Our results indicated cytoprotective roles for ORP150 in rat heart and suggested a therapeutic role for the protein both in preventing cardiomyocyte death and in preserving contractile function after ischemic damage. (c) 2005 Elsevier Ltd. All rights reserved.