Hypoxic preconditioning up-regulates DJ-1 protein expression in rat heart-derived H9c2 cells through the activation of extracellular-regulated kinase 1/2 pathway
Hypoxic preconditioning up-regulates DJ-1 protein expression in rat heart-derived H9c2 cells through the activation of extracellular-regulated kinase 1/2 pathway
复制标题
缺氧预处理通过激活细胞外调节激酶1/2通路上调大鼠心脏源性H9c2细胞中DJ-1蛋白表达
DOI：
10.1007/s11010-012-1414-8
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发表时间：
2012-11-01
影响因子：
4.3
通讯作者：
He, Ming
中科院分区：
文献类型：
--
作者：
Lu, Hai-Shan;Chen, He-Ping;He, Ming
心肌预适应是一种强大的现象，可以减轻缺血/再灌注引起的氧化应激并引起延迟的心脏保护。其机制涉及细胞内信号通路的激活和保护性抗氧化蛋白的上调。DJ-1蛋白是一种多功能细胞内蛋白，在减缓氧化应激、促进细胞存活等方面发挥着重要作用。在本研究中，我们研究了DJ-1是否在缺氧预处理（HP）后期上调，并且DJ-1的上调是通过细胞外调节激酶1/2 （ERK1/2）信号通路介导的。大鼠心脏源性H9c2细胞暴露于HP。24h后进行缺氧/复氧（H/R）处理，适当测定细胞活力、乳酸脱氢酶（LDH）、细胞内活性氧（ROS）、ERK1/2磷酸化水平和DJ-1蛋白水平。结果表明，HP能有效抑制H/ r诱导的活力损失和LDH泄漏。HP促进ERK1/2活化，上调DJ-1蛋白表达，抑制H/R诱导的ROS升高。然而，当ERK1/2磷酸化被U0126特异性抑制时，HP期间发生的DJ-1表达增加几乎被完全消除，从而消除HP诱导的延迟心脏保护作用，HP对H/ r诱导的氧化应激的抑制作用也被逆转。此外，通过siRNA敲除DJ-1可以减弱HP诱导的延迟心脏保护。我们的数据表明，HP可以通过erk1 /2依赖的信号通路上调DJ-1蛋白的表达。重要的是，DJ-1可能参与HP对H/R损伤的延迟心脏保护作用。
Myocardial preconditioning is a powerful phenomenon that can attenuate ischemia/reperfusion-induced oxidant stress and elicit delayed cardioprotection. Its mechanisms involve activation of intracellular signaling pathways and up-regulation of the protective antioxidant proteins. DJ-1 protein, as a multifunctional intracellular protein, plays an important role in attenuating oxidant stress and promoting cell survival. In the present study, we investigated whether DJ-1 is up-regulated during the late phase of hypoxic preconditioning (HP) and the up-regulation of DJ-1 is mediated by extracellular-regulated kinase 1/2 (ERK1/2) signaling pathway. Rat heart-derived H9c2 cells were exposed to HP. Twenty-four hours later cells were subjected to hypoxia/reoxygenation (H/R) and then cell viability, lactate dehydrogenase (LDH), intracellular reactive oxygen species (ROS), ERK1/2 phosphorylation, and DJ-1 protein were measured appropriately. The results showed that HP efficiently attenuated H/R-induced viability loss and LDH leakage. In addition, HP promoted ERK1/2 activation, up-regulated DJ-1 protein expression, inhibited H/R induced the elevation of ROS. However, when ERK1/2 phosphorylation was specifically inhibited by U0126, the increase in DJ-1 expression occurring during HP was almost completely abolished and, as a result, the delayed cardioprotection induced by HP was abolished, and the inhibitory effect of HP on H/R-induced oxidant stress was also reversed. Furthermore, knocking down DJ-1 by siRNA attenuated the delayed cardioprotection induced by HP. Our data indicate that HP can up-regulate DJ-1 protein expression through the ERK1/2-dependent signaling pathway. Importantly, DJ-1 might be involved in the delayed cardioprotective effect of HP against H/R injury.