Redox activation of Ref-1 potentiates cell survival following myocardial ischemia reperfusion injury
Redox activation of Ref-1 potentiates cell survival following myocardial ischemia reperfusion injury
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DOI：
10.1016/j.freeradbiomed.2007.04.025
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发表时间：
2007-08-01
影响因子：
7.4
通讯作者：
Das, Dipak K.
中科院分区：
文献类型：
--
作者：
Gurusamy, Narasimman;Malik, Gautam;Das, Dipak K.
最近的一项研究表明，心脏适应性可以加强硫氧还蛋白-1(Trx-1)的移位，使其进入细胞核，然后与Ref-1相互作用，产生生存信号。在这里，我们提出的证据表明，这种适应也导致Ref-1的核转位，当心脏被反义Ref-1预处理时，Ref-1的核转位几乎完全被抑制，反义Ref-1也取消了心脏保护性适应性反应。当用Ref-1免疫沉淀左心室核提取液时，发现大量的NF-kappa B和Nrf2与Ref-1相关。反义Ref-1显著抑制了Ref-1-NF-kappa B和Ref-1-Nrf2的相互作用。然而，核抽提物与核因子kappa B的免疫沉淀表明，在适应的心脏中，Trx-1与核因子kappaB的结合增加，这再次被反义Ref-1显著阻断。NRF2也与核因子kappaB相关；然而，这种关联似乎不依赖于Ref-1。相反，心肌对缺血的适应性抑制了缺血再灌注引起的核内Nrf2的丢失，这一作用可被反义Ref-1抑制。Ref-1的核转位和激活似乎产生了存活信号，Akt的磷酸化增加可以被反义Ref-1抑制。最后，共聚焦显微镜证实了免疫印迹的结果，清楚地显示了Ref-1的核转位以及Ref-1与核因子kappaB在适应心脏中的核3D共定位，并显示了Ref-1的反义抑制作用。我们的结果表明，PC通过Akt的磷酸化导致核转位和Ref-1的激活来增强生存信号，而NF-kappa B与Ref-1、Trx-1和Nrf2之间的显著相互作用似乎调节了Ref-1诱导的生存信号。(C)2007 Elsevier Inc.保留所有权利。
A recent study showed that cardiac adaptation could potentiate translocation of thioredoxin-1 (Trx-1) into the nucleus, which then interacted with Ref-1, resulting in a survival signal. Here, we present evidence that such adaptation also causes nuclear translocation of Ref-1, which is almost completely inhibited when the hearts were pretreated with antisense Ref-1 that also abolished the cardioprotective adaptive response. Significant amounts of NF kappa B and Nrf2 were found to be associated with Ref-1 when the nuclear extract obtained from the left ventricle was immunoprecipitated with Ref-1. Such Ref-1-NF kappa B and Ref-1-Nrf2 interactions were significantly inhibited with antisense Ref-1. However, immunoprecipitation of nuclear extract with NF kappa B showed that the association of Trx-1 with NF kappa B is increased in the adapted heart, which was again significantly blocked by antisense Ref-1. Nrf2 was also associated with NF kappa B; however, such association appeared to be independent of Ref-1. In contrast, myocardial adaptation to ischemia inhibited the ischemia reperfusion-induced loss of Nrf2 from the nucleus, which was inhibited by antisense Ref-1. The nuclear trans location and activation of Ref-1 appeared to generate a survival signal as evidenced by the increased phosphorylation of Akt that was inhibited with antisense Ref-1. Finally, confocal microscopy confirmed the results of immunoblotting, clearly showing the nuclear translocation of Ref-1 and nuclear 3D colocalization of Ref-1 with NF kappa B in the adapted heart and its inhibition with antisense Ref-1. Our results show that PC potentiates a survival signal through the phosphorylation of Akt by causing nuclear translocation and activation of Ref-1, where significant interaction among NF kappa B and Ref-1, Trx-1, and Nrf2 appears to regulate Ref-1 -induced survival signal. (C) 2007 Elsevier Inc. All rights reserved.