Glucose reintroduction triggers the activation of Nrf2 during experimental ischemia reperfusion
Glucose reintroduction triggers the activation of Nrf2 during experimental ischemia reperfusion
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DOI：
10.1007/s11010-012-1300-4
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发表时间：
2012-07-01
影响因子：
4.3
通讯作者：
Leonard, Martin O.
中科院分区：
文献类型：
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作者：
Crean, Daniel;Felice, Luca;Leonard, Martin O.
再灌注导致氧气、葡萄糖和其他受限成分快速重新引入缺血组织。它不仅带来了细胞生存所必需的成分，还带来了氧化应激和细胞损伤的爆发。在这项研究中，我们的主要目的是研究葡萄糖作为再灌注时转录因子 NF-E2 相关因子 2 (Nrf2) 激活的决定因素以及下游抗氧化剂 NADPH 依赖性还原酶的表达。将肾上皮 HK-2 细胞暴露于氧气和葡萄糖消耗后重新引入的环境中，导致 Nrf2 蛋白的核转位以依赖于葡萄糖的方式增加。观察到 Nrf2 依赖性基因 NAD(P)H 脱氢酶醌 1 (NQO1) 的激活和诱导在 5 mM 葡萄糖浓度下达到最大。在这些条件下对 siRNA 靶向细胞的 mRNA 进行微阵列分析，揭示了 Nrf2 依赖性 NADPH 依赖性还原酶 NQO1、醛酮还原酶家族 1、成员 C1-3 和脱氢酶/还原酶（SDR 家族）成员 2 (DHRS2) 的 Nrf2 依赖性表达，所有基因均被证明可以防止氧化应激介导的细胞损伤。此外，NQO1和DHRS2 mRNA水平在葡萄糖重新引入时特异性上调，并且在小鼠肾蒂钳夹的体内缺血再灌注损伤模型中也增加。总之，我们证明葡萄糖耗尽后重新引入葡萄糖会激活 Nrf2 和 Nrf2 调节 NADPH 依赖性还原酶的表达。我们认为这些发现代表了一种先前未报道的 Nrf2 激活机制，作为 IRI 中的细胞保护途径。
Reperfusion results in a rapid reintroduction of oxygen, glucose, and other restricted components to an ischemic tissue. It brings with it not only the necessary components for cell survival but also a burst of oxidative stress and cellular damage. In this study, our primary aims were to investigate glucose as a determining factor for the activation of the transcription factor NF-E2-related factor 2 (Nrf2) upon reperfusion and the expression of downstream anti-oxidant NADPH-dependent reductases. Exposure of renal epithelial HK-2 cells to oxygen and glucose reintroduction after depletion resulted in an increase in nuclear translocation of Nrf2 protein in a manner dependent upon glucose. This activation and the induction of the Nrf2-dependent gene NAD(P)H dehydrogenase, quinone 1 (NQO1) was observed to be maximum at a concentration of 5 mM glucose. Microarray analysis of mRNA from siRNA targeted cells under these conditions revealed the Nrf2-dependent expression of NADPH-dependent reductase enzymes NQO1, Aldo-keto reductase family 1, members C1-3 and dehydrogenase/reductase (SDR family) member 2 (DHRS2), all genes demonstrated to protect against oxidative stress-mediated cellular injury. In addition, NQO1 and DHRS2 mRNA levels were specifically upregulated on glucose reintroduction and were also increased in an in vivo ischemia reperfusion injury model of murine renal pedicle clamping. In conclusion, we demonstrate that glucose reintroduction after depletion activates Nrf2 and Nrf2 regulated NADPH-dependent reductase expression. We suggest these findings represent a previously unreported mechanism for the activation of Nrf2 as a cytoprotective pathway in IRI.