NF-E2-related factor-2 mediates neuroprotection against mitochondrial complex I inhibitors and increased concentrations of intracellular calcium in primary cortical neurons
NF-E2-related factor-2 mediates neuroprotection against mitochondrial complex I inhibitors and increased concentrations of intracellular calcium in primary cortical neurons
复制标题
DOI：
10.1074/jbc.m305204200
复制
发表时间：
2003-09-26
影响因子：
4.8
通讯作者：
Johnson, JA
中科院分区：
文献类型：
--
作者：
Lee, JM;Shih, AY;Johnson, JA
NF-E2 相关因子 2 (Nrf2) 通过称为抗氧化响应元件 (ARE) 的增强子序列调节 II 相解毒酶和抗氧化蛋白的基因表达。在这项研究中，我们证明 Nrf2 通过协调上调 ARE 驱动基因来保护含有星形胶质细胞（大约 10%）和神经元（大约 90%）的混合原代神经元培养物中的神经元。与Nrf2(+/+)神经元相比，该混合培养系统中的Nrf2(-/-)神经元对线粒体毒素(1-甲基-4-苯基-1,2,5,6-四氢吡啶或鱼藤酮)诱导的细胞凋亡更敏感。为了了解这种观察到的差异敏感性的潜在机制，我们使用寡核苷酸微阵列比较了基因表达谱。微阵列数据显示，与 Nrf2(-/-) 神经元培养物相比，Nrf2(+/+) 神经元培养物中编码解毒酶、抗氧化蛋白、钙稳态蛋白、生长因子、神经元特异性蛋白和信号分子的基因表达水平更高。正如从微阵列数据预测的那样，Nrf2(-/-)神经元确实更容易受到离子霉素和2,5-二(叔丁基)-1,4-氢醌诱导的细胞内钙增加的细胞毒性作用的影响。最后，腺病毒载体介导的Nrf2过表达恢复了Nrf2(-/-)神经元培养物中ARE驱动的基因表达，并将Nrf2(+/+)神经元从鱼藤酮或离子霉素诱导的细胞死亡中拯救出来。总而言之，这些发现表明 Nrf2 在保护神经元免受毒性损伤方面发挥着重要作用。
NF-E2-related factor-2 (Nrf2) regulates the gene expression of phase II detoxification enzymes and antioxidant proteins through an enhancer sequence referred to as the antioxidant-responsive element ( ARE). In this study, we demonstrate that Nrf2 protects neurons in mixed primary neuronal cultures containing both astrocytes (similar to 10%) and neurons (similar to 90%) through coordinate up-regulation of ARE- driven genes. Nrf2(-/-) neurons in this mixed culture system were more sensitive to mitochondrial toxin ( 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine or rotenone)-induced apoptosis compared with Nrf2(+/+) neurons. To understand the underlying mechanism of this observed differential sensitivity, we compared the gene expression profiles using oligonucleotide microarrays. Microarray data showed that Nrf2(+/+) neuronal cultures had higher expression levels of genes encoding detoxification enzymes, antioxidant proteins, calcium homeostasis proteins, growth factors, neuron-specific proteins, and signaling molecules compared with Nrf2(-/-) neuronal cultures. As predicted from the microarray data, Nrf2(-/-) neurons were indeed more vulnerable to the cytotoxic effects of ionomycin- and 2,5-di( t-butyl)-1,4-hydroquinone-induced increases in intracellular calcium. Finally, adenoviral vector-mediated overexpression of Nrf2 recovered ARE- driven gene expression in Nrf2(-/-) neuronal cultures and rescued Nrf2(+/+) neurons from rotenone- or ionomycin- induced cell death. Taken together, these findings suggest that Nrf2 plays an important role in protecting neurons from toxic insult.