Identification of the NF-E2-related Factor-2-dependent Genes Conferring Protection against Oxidative Stress in Primary Cortical Astrocytes Using Oligonucleotide Microarray Analysis

Abstract

The antioxidant responsive element (ARE) mediates transcriptional regulation of phase II detoxification enzymes and antioxidant proteins such as NAD(P)H:quinone oxidoreductase (NQO1), glutathione S-transferases, and glutamate-cysteine ligase. In this study, we demonstrate that NF-E2-related factor-2 (Nrf2) plays a major role in transcriptional activation of ARE-driven genes and identify Nrf2-dependent genes by oligonucleotide microarray analysis using primary cortical astrocytes from Nrf2 ؉/؉ and Nrf2 ؊/؊ mice. Nrf2 ؊/؊ astrocytes had decreased basal NQO1 activity and no induction by tert-butylhydroquinone compared with Nrf2 ؉/؉ astrocytes. Similarly, both basal and induced levels of human NQO1-ARE-luciferase expression in Nrf2 ؊/؊ astrocytes were significantly lower than in Nrf2 ؉/؉ astrocytes. Furthermore, human NQO1-ARE-luciferase expression in Nrf2 ؊/؊ astrocytes was restored by overexpression of Nrf2, whereas ARE activation in Nrf2 ؉/؉ astrocytes was completely blocked by dominantnegative Nrf2. In addition, we observed that Nrf2-dependent genes protected primary astrocytes from H 2 O 2 -or platelet-activating factor-induced apoptosis. In support of these observations, we identified Nrf2-dependent genes encoding detoxification enzymes, glutathione-related proteins, antioxidant proteins, NADPHproducing enzymes, and anti-inflammatory genes using oligonucleotide microarrays. Proteins within these functional categories are vital to the maintenance and responsiveness of a cell defense system, suggesting that an orchestrated change in gene expression via Nrf2 and the ARE gives a synergistic protective effect against oxidative stress.

Astroglia as a cellular target for neuroprotection and treatment of neuro‐psychiatric disorders

“…Hence, the Nrf2‐ARE pathway is considered a high‐value therapeutic target (de Vries et al, 2008; Johnson and Johnson, 2015; van Muiswinkel and Kuiperij, 2005). It is preferentially activated in astrocytes while neurons largely depend on astrocytes for the antioxidant defense (Kraft, Johnson, & Johnson, 2004; Lee, Calkins, Chan, Kan, & Johnson, 2003; Shih et al, 2003). Therefore, unsurprisingly, many studies report that activation of the Nrf2 pathway in astrocytes is neuroprotective (Calkins, Vargas, Johnson, & Johnson, 2010; Chen et al, 2009; Gan, Vargas, Johnson, & Johnson, 2012; Vargas, Johnson, Sirkis, Messing, & Johnson, 2008).…”

Flunarizine induces Nrf2-mediated transcriptional activation of heme oxygenase-1 in protection of auditory cells from cisplatin

“…The primary astrocyte of Nrf2 À/À mice is also more susceptible to oxidative stress and inflammation than that of Nrf2 þ / þ mice. 6 Leung et al 7 showed that deficiency of Nrf2 results in early embryonic lethality with severe oxidative stress. These observations, collectively, imply that Nrf2 is a master regulator of ARE-driven transcriptional activation for antioxidant genes in maintaining the homeostasis of Redox status within cells.…”

Dietary leucine improves flesh quality and alters mRNA expressions of Nrf2-mediated antioxidant enzymes in the muscle of grass carp ( Ctenopharyngodon idella )

“…The transcription factor Nrf2 is a central protein that regulates the transcription of multiple antioxidant enzyme genes in mice primary cortical astrocytes (Lee et al, 2003). In mice, the up-regulation of Nrf2 expression in the lung induced greater expression of Nrf2 target antioxidant genes, such as CAT and GPx (Blake et al, 2010).…”