Nrf2-induced antioxidant protection: a promising target to counteract ROS-mediated damage in neurodegenerative disease?

Neurodegenerative diseases share various pathological features, such as accumulation of aberrant protein aggregates, microglial activation, and mitochondrial dysfunction. These pathological processes are associated with generation of reactive oxygen species (ROS), which cause oxidative stress and subsequent damage to essential molecules, such as lipids, proteins, and DNA. Hence, enhanced ROS production and oxidative injury play a cardinal role in the onset and progression of neurodegenerative disorders. To maintain a proper redox balance, the central nervous system is endowed with an antioxidant defense mechanism consisting of endogenous antioxidant enzymes. Expression of most antioxidant enzymes is tightly controlled by the antioxidant response element (ARE) and is activated by nuclear factor E2-related factor 2 (Nrf2). In past years reports have highlighted the protective effects of Nrf2 activation in reducing oxidative stress in both in vitro and in vivo models of neurodegenerative disorders. Here we provide an overview of the involvement of ROS-induced oxidative damage in Alzheimer's disease, Parkinson's disease, and Huntington's disease and we discuss the potential therapeutic effects of antioxidant enzymes and compounds that activate the Nrf2-ARE pathway.     

Nrf2-ARE信号通路在神经退行性疾病中的作用研究进展

神经退行性疾病(Neurodegenerative disease)是一类以神经元退行性病变为基础的慢性、进行性、不可逆的神经系统疾病的总称,主要包括阿尔茨海默病(Alzheimer's disease,AD)、帕金森病(Parkinson's disease,PD)、亨廷顿舞蹈病(Huntington disease,HD)、肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)、脊髓肌萎缩症(spinal muscular atrophy,SMA)、不同类型脊髓小脑共济失调(spinal cerebella ataxias,SCA)等。其病因和发病机制十分复杂,其中,氧化应激学说近年来受到了人们的广泛关注和普遍认可。而研究表明,Nrf2-ARE信号通路是体内抗氧化应答机制中最重要的通路之一,其能对氧化应激导致的神经细胞损伤产生保护作用,即阻止神经细胞的病变和凋亡,进而延缓神经退行性疾病的发生发展,因而有望成为神经退行性疾病的有效治疗靶标。本文就Nrf2-ARE信号通路结构特点及Nrf2-ARE信号通路在神经退行性疾病中的作用研究进展作一综述。     

Effects of different exercise durations on Keap1-Nrf2-ARE pathway activation in mouse skeletal muscle

Abstract

The purpose of this study was to investigate the effects of acute exercise stress on the nuclear factor-erythroid2 p45-related factor 2 (Nrf2)/antioxidant response element (ARE) transactivation, Kelch-like ECH-associated protein 1 (Keap1) cytosolic protein and Nrf2 nucleoprotein expressions, Nrf2 target genes mRNA expressions, and glutathione redox (GSH/GSSG) ratio level; with a particular focus on the changes in Keap1-Nrf2-ARE pathway activation following different durations of exercise. Wild-type mice (C57BL/6J, two months old) were separated into one-hour and six-hour treadmill running groups, as well as a non-exercise control group (n = 10 in each group). Measurements of Nrf2/ARE transactivation, Nrf2 nucleoprotein expressions, Keap1 cytosolic protein expression, Nrf2 target genes’ mRNA expressions (superoxide dismutase-1 [SOD1], superoxide dismutase-2 [SOD2], γ-glutamyl cysteine ligase-modulatory [GCLm], γ-glutamyl cysteine ligase-catalytic [GCLc], glutathione reductase [GR], glutathione peroxidase-1 [Gpx1], catalase [CAT], and hemoxygenase-1 [Ho-1]), and GSH/GSSG ratio were carried out immediately after exercise. The results showed significant increases in Keap1-Nrf2-ARE pathway activation and the mRNA expressions of six measured enzymes in skeletal muscle after six hours of exercise; while in the one-hour exercise group, there was no change in Keap1-Nrf2-ARE pathway activation and only two enzymes’ mRNA expressions were increased. It is suggested that the changes in Keap1-Nrf2-ARE pathway activation and its target genes’ mRNA expressions were dependent on the exercise duration, with longer duration associated with higher responses.     

Biogenic nanoselenium particles activate Nrf2-ARE pathway by phosphorylating p38, ERK1/2, and AKT on IPEC-J2 cells

As the intestinal epithelium is vulnerable to oxidative stress because of frequent enterocyte renewal and continuous exposure to exogenous agents, it is meaningful to figure out how the epithelial cells exert antioxidant function. We previously synthesized a novel biogenic nanoselenium (BNS) particles and proved that BNS could effectively improve intestinal antioxidative function through activating Nrf2-ARE pathway. The objective of the present study was to investigate the mechanism by which BNS activate Nrf2-ARE pathway on the physiological function of intestinal epithelial cells. In the present study, we demonstrated that treatment of IPEC-J2 cells with BNS particles not only elevated the levels of downstream proteins of nuclear factor (erythroid-derived-2)-like 2 (Nrf2) such as heme oxygenase-1 and NQO-1 in a time-dependent manner which started to weaken at 12 hr after treatment but also significantly activated Nrf2, mitogen-activated protein kinase (MAPK), and protein kinase B (AKT) pathway in a time-dependent manner within 24 hr. BNS particles significantly increased the content of phosphorylated-Nrf2, without evident influence on the level of Kelch-like ECH-associated protein 1 (Keap1). Moreover, BNS also induced the activation of p38, extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase, and AKT while phosphorylating Nrf2. Using specific protein kinase inhibitors, we found that the Nrf2-phosphorylating and antioxidative effects of BNS particles were abolished when p38, ERK1/2, and AKT were significantly inhibited. Overall, our data demonstrated that BNS particles activated Nrf2-ARE pathway through p38, ERK1/2, and AKT mediated-phosphorylation of Nrf2 to improve the antioxidant function of intestinal epithelial cells     

转录因子Nrf2在肝脏疾病中的作用

NF-E2相关因子2(nuclear erythroid 2-related factor 2,Nrf2)是一种能调节肝脏中大量解毒和抗氧化防御基因表达的重要转录因子.氧化应激与各种形式的肝损伤有密切的关系.Nrf2由亲电体压力或氧化应激激活,并通过结合抗氧化反应元件(antioxidant response element,ARE)诱导其靶基因,从而对细胞产生保护作用.因此,Nrf2通路在肝脏疾病中的作用已被深入研究.多种动物模型研究结果表明,Nrf2通路通过靶基因表达,在对抗病毒性肝炎、药物性肝损伤、酒精性肝病、非酒精性脂肪肝及肝癌方面表现出了不同的生物功能.根据Nrf2及其信号通路在对抗肝损伤中产生保护作用的相关文献,本文综述并讨论了其作为治疗肝损伤的药物作用靶点方面可能的应用前景.     

Isolation, identification, and biological evaluation of Nrf2-ARE activator from the leaves of green perilla (Perilla frutescens var. crispa f. viridis)

The nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway is a cellular defense system against oxidative stress. Activation of this pathway increases expression of antioxidant enzymes. Epidemiological studies have demonstrated that the consumption of fruits and vegetables is associated with reduced risk of contracting a variety of human diseases. The aim of this study is to find Nrf2-ARE activators in dietary fruits and vegetables. We first attempted to compare the potency of ARE activation in six fruit and six vegetables extracts. Green perilla (Perilla frutescens var. crispa f. viridis) extract exhibited high ARE activity. We isolated the active fraction from green perilla extract through bioactivity-guided fractionation. Based on nuclear magnetic resonance and mass spectrometric analysis, the active ingredient responsible for the ARE activity was identified as 2',3'-dihydroxy-4',6'-dimethoxychalcone (DDC). DDC induced the expression of antioxidant enzymes, such as γ-glutamylcysteine synthetase (γ-GCS), NAD(P)H: quinone oxidoreductase-1 (NQO1), and heme oxygenase-1. DDC inhibited the formation of intracellular reactive oxygen species and the cytotoxicity induced by 6-hydroxydopamine. Inhibition of the p38 mitogen-activated protein kinase pathway abolished ARE activation, the induction of γ-GCS and NQO1, and the cytoprotective effect brought about by DDC. Thus, this study demonstrated that DDC contained in green perilla enhanced cellular resistance to oxidative damage through activation of the Nrf2-ARE pathway.     

Enhancement of dopaminergic activity and region-specific activation of Nrf2-ARE pathway by intranasal supplements of testosterone propionate in aged male rats

The potential influence of intranasal testosterone propionate (InTP) supplements on mesodopaminergic system in aged male rats was investigated by analyzing the exploratory and motor behaviors as well as dopamine neurobiochemical indices. Meanwhile, oxidative stress parameters and pathway of nuclear factor erythroid 2-related factor 2 (Nrf2)-binding antioxidant response elements (Nrf2-ARE) were examined to check whether the Nrf2-ARE pathway was involved in the InTP-induced alteration of mesodopaminergic system in aged male rats. The exploratory and motor behavioral deficits, as well as the reduced expression of dopamine, tyrosine hydroxylase, and dopamine transporter, which indicated the declined activity of mesodopaminergic system, were ameliorated in rats administered with 12-week InTP. The results indicated that chronic InTP supplements could effectively influence the brain function activity in a way opposite to the effect of aging on the mesodopaminergic system of rats. The increased levels of Nrf2, heme oxygenase-1, and NAD(P)H:quinone oxidoreductase-1 in the substantia nigra and ventral tegmental area, but not in the hippocampus of InTP-administered aged male rats, indicated that the ameliorative effect of InTP supplements on mesodopaminergic system might be related to the region-specific activation of the Nrf2-ARE pathway.     

Brazilian red propolis extract enhances expression of antioxidant enzyme genes in vitro and in vivo

ABSTRACT

Brazilian red propolis reportedly has reactive oxygen species (ROS) scavenging effects in vitro, but the cellular mechanisms remain unclear. In the present study, the effects of an ethanol extract of Brazilian red propolis (EERP) on the Nrf2-ARE intracellular antioxidant pathway were examined in vitro and in vivo. EERP and its constituents transactivated the reporter gene through the ARE sequence and enhanced the expression of Nrf2-regulated genes in HEK293 cells. It also increased Nrf2 protein in the nucleus, which was partially inhibited by kinase inhibitors. Furthermore, EERP suppressed ROS generation and cytotoxicity induced by tert-butyl hydroperoxide. In vivo, orally administered EERP increased the expression of Nrf2-regulated genes in mice liver. These results suggest that EERP is a potential resource for preventing oxidative stress-related diseases as an Nrf2 inducer.