Nuclear factor E2-related factor 2-dependent antioxidant response element activation by tert-butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult
Nuclear factor E2-related factor 2-dependent antioxidant response element activation by tert-butylhydroquinone and sulforaphane occurring preferentially in astrocytes conditions neurons against oxidative insult
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DOI：
10.1523/jneurosci.3817-03.2004
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发表时间：
2004-02-04
影响因子：
5.3
通讯作者：
Johnson, JA
中科院分区：
文献类型：
--
作者：
Kraft, AD;Johnson, DA;Johnson, JA
转录因子核因子E2相关因子2（Nrf 2）与神经细胞中的抗氧化反应元件（ARE）结合，导致诱导一系列基因，这些基因可以协调针对多种氧化应激源的保护性反应。在这项研究中，叔丁基对苯二酚（tBHQ）和萝卜硫素被用作该途径的激活剂。与先前的研究一致，ARE报告小鼠的原代皮层培养物的处理揭示了星形胶质细胞中的选择性启动子活性。这种激活保护神经元免受过氧化氢和非兴奋性毒性谷氨酸毒性。tBHQ处理来自Nrf 2敲除动物的培养物既不导致ARE活化也不导致神经保护。通过用复制缺陷型腺病毒（ad）感染重新引入Nrf 2，遗传反应和神经保护都得到了拯救。相反，用编码显性阴性（DN）Nrf 2的腺病毒（ad-DN-Nrf 2）感染或用选择性磷脂酰肌醇-3激酶抑制剂LY 294002预处理抑制tBHQ介导的启动子应答和相应的神经保护。有趣的是，腺病毒感染对星形胶质细胞的选择性高于神经元。为了揭示ARE激活引起的某些细胞类型特异性变化，在tBHQ处理之前，用编码绿色荧光蛋白（GFP）的腺病毒（ad-GFP）或ad-DN-Nrf 2（含有GFP）感染培养物。然后使用细胞分选技术从未感染的神经元群体中分离出富含胶质细胞的GFP感染细胞群体。微阵列分析被用来评估潜在的神经胶质细胞与神经元特异性贡献的神经保护作用的ARE激活和Nrf 2依赖。引人注目的是，tBHQ处理后神经元基因表达的变化依赖于星形胶质细胞中的Nrf 2活性。这表明Nrf 2依赖性遗传变化改变神经元-胶质细胞相互作用，从而产生神经保护作用。
Binding of the transcription factor nuclear factor E2-related factor 2 (Nrf2) to the antioxidant response element (ARE) in neural cells results in the induction of a battery of genes that can coordinate a protective response against a variety of oxidative stressors. In this study, tert-butylhydroquinone (tBHQ) and sulforaphane were used as activators of this pathway. Consistent with previous studies, treatment of primary cortical cultures from ARE reporter mice revealed selective promoter activity in astrocytes. This activation protected neurons from hydrogen peroxide and nonexcitotoxic glutamate toxicity. tBHQ treatment of cultures from Nrf2 knock-out animals resulted in neither ARE activation nor neuroprotection. By reintroducing Nrf2 via infection with a replication-deficient adenovirus (ad), both the genetic response and neuroprotection were rescued. Conversely, infection with adenovirus encoding dominant-negative (DN) Nrf2 (ad-DN-Nrf2) or pretreatment with the selective phosphatidylinositol-3 kinase inhibitor LY294002 inhibited the tBHQ-mediated promoter response and corresponding neuroprotection. Interestingly, the adenoviral infection showed a high selectivity for astrocytes over neurons. In an attempt to reveal some of the cell type-specific changes resulting from ARE activation, cultures were infected with adenovirus encoding green fluorescent protein (GFP) (ad-GFP) or ad-DN-Nrf2 (containing GFP) before tBHQ treatment. A glia-enriched population of GFP-infected cells was then isolated from a population of uninfected neurons using cell-sorting technology. Microarray analysis was used to evaluate potential glial versus neuron-specific contributions to the neuroprotective effects of ARE activation and Nrf2 dependence. Strikingly, the change in neuronal gene expression after tBHQ treatment was dependent on Nrf2 activity in the astrocytes. This suggests that Nrf2-dependent genetic changes alter neuron-glia interactions resulting in neuroprotection.