Nrf2抗氧化的分子调控机制

Nrf2是调控细胞氧化应激反应的重要转录因子,同时也是维持细胞内氧化还原稳态的中枢调节者。Nrf2通过诱导调控一系列抗氧化蛋白的组成型和诱导型表达,可以减轻活性氧和亲电体引起的细胞损伤,使细胞处于稳定状态,维持机体氧化还原动态平衡。本研究为了从分子层面深入探讨剖析Nrf2发挥抗氧化功能的作用机制,通过查找阅读大量相关文献并进行整理归纳,最终从Nrf2的结构与激活、Nrf2抗氧化功能以及Nrf2抗氧化的分子调控机制三个方面进行了概述分析。其中在对Nrf2抗氧化的分子调控机制的探讨部分,既探析了对Nrf2起激活作用的相关调节因子的作用机制,又分析了Nrf2被激活后对其下游多种抗氧化因子及谷胱甘肽氧化还原系统的诱导调控机制,以期较深入了解Nrf2抵抗机体氧化应激损伤作用及其抗氧化分子调控机制。     

Gualou Guizhi Granule Protects Against Oxidative Injury by Activating Nrf2/ARE Pathway in Rats and PC12 Cells

Stroke involves numerous pathophysiological processes and oxidative stress is considered as a main cellular event in its pathogenesis. The nuclear factor erythroid-2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway plays a key role in inducing phase II detoxifying enzymes and antioxidant proteins and is now considered as a interesting therapeutic target for the treatment of stroke. The objective of this study is to investigate the protective effect of Gualou Guizhi granule (GLGZG) against oxidative stress and explore the protective mechanism of the Nrf2/ARE pathway. In vivo, administration of GLGZG in a rat model of focal cerebral ischemia significantly suppressed oxidative injury by increasing the activity of superoxide dismutase and glutathione level and decreasing reactive oxygen species and malondialdehyde levels. Western blot analysis showed that GLGZG induced nuclear translocation of Nrf2, and combined with real-time PCR results, which indicated that GLGZG up-regulated the Nrf2/ARE pathway. In addition, in cultured PC12 cells, GLGZG protected against H₂O₂ induced oxidative injury and activated the Nrf2/ARE pathway. All the results demonstrated that GLGZG in the management of cerebral ischemia and H₂O₂ induced oxidative injury may be associated with activation of Nrf2/ARE signaling pathway.     

Hypochlorous acid alters bronchial epithelial cell membrane properties and prevention by extracellular glutathione.

In chronic inflammatory diseases of the airways, such as cystic fibrosis, hypochlorous acid (HOCl) generated by neutrophils is involved in airway injury. We examined the effects of HOCl on 16HBE14o- bronchial epithelial cells by bolus addition or by generation with glucose oxidase plus myeloperoxidase. HOCl produced both carbonyl formation of a discreet number of proteins and modification of surface targets that were recognized by an antibody raised against HOCl-modified protein. Bolus or enzymatically generated HOCl decreased transepithelial resistance, but surprisingly bolus HOCl also increased short-circuit current. Glutathione in lung epithelial lining fluid is an excellent scavenger of HOCl; however, glutathione content is lower in cystic fibrosis epithelial lining fluid due to deficient glutathione transport to the apical side of bronchial-tracheal epithelial cells (Gao L, Kim KJ, Yankaskas JR, and Forman HJ. Am J Physiol Lung Cell Mol Physiol 277: L113-L118, 1999). We found that alteration of the GSH content of apical fluid above 16HBE14o- cells was protective because all HOCl-induced changes were delayed or eliminated by exogenous glutathione within the physiological range. Extrapolating this to cystic fibrosis suggests that HOCl can alter cell function without destruction but that elevating glutathione could be protective.     

Nrf2在阿尔茨海默病中的研究现状

阿尔茨海默病(Alzheimer's disease,AD)是最常见的神经系统变性疾病,主要病理特征为细胞外老年斑(senile plaques,SP)和细胞内神经原纤维缠结(neurofibrillary tangles,NFT)形成.但其发病机制不清,涉及多种病理学变化如炎症反应、氧化应激、线粒体功能障碍、细胞凋亡以及突触功能障碍等.核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)是经典的调控机体抗氧化应激反应的核转录因子.Nrf2激活后诱导抗氧化蛋白的表达,提高机体的抗氧化应激能力.随着Nrf2抗氧化应激作用研究的深入,发现Nrf2不仅能够通过抗氧化应激延缓AD的发生发展,且在AD的病理性沉积物的清除、抗炎、抗凋亡、神经营养等方面扮演着重要的角色.近年来,由于多种针对单一靶点的抗AD药物临床试验的失败,有学者提出Nrf2可能是实现AD多靶点疗法的重要因子.因此,本文对Nrf2在AD中的研究现状做一综述,为寻找治疗AD潜在的生物学靶点提供理论依据.     

Human alveolar epithelial cell injury induced by cigarette smoke

Background

Cigarette smoke (CS) is a highly complex mixture and many of its components are known carcinogens, mutagens, and other toxic substances. CS induces oxidative stress and cell death, and this cell toxicity plays a key role in the pathogenesis of several pulmonary diseases.

Methodology/Principal Findings

We studied the effect of cigarette smoke extract (CSE) in human alveolar epithelial type I-like (ATI-like) cells. These are isolated type II cells that are differentiating toward the type I cell phenotype in vitro and have lost many type II cell markers and express type I cell markers. ATI-like cells were more sensitive to CSE than alveolar type II cells, which maintained their differentiated phenotype in vitro. We observed disruption of mitochondrial membrane potential, apoptosis and necrosis that were detected by double staining with acridine orange and ethidium bromide or Hoechst 33342 and propidium iodide and TUNEL assay after treatment with CSE. We also detected caspase 3 and caspase 7 activities and lipid peroxidation. CSE induced nuclear translocation of Nrf2 and increased expression of Nrf2, HO-1, Hsp70 and Fra1. Moreover, we found that Nrf2 knockdown sensitized ATI-like cells to CSE and Nrf2 overexpression provided protection against CSE-induced cell death. We also observed that two antioxidant compounds N-acetylcysteine and trolox protected ATI-like cells against injury by CSE.

Conclusions

Our study indicates that Nrf2 activation is a major factor in cellular defense of the human alveolar epithelium against CSE-induced toxicity and oxidative stress. Therefore, antioxidant agents that modulate Nrf2 would be expected to restore antioxidant and detoxifying enzymes and to prevent CS-related lung injury and perhaps lessen the development of emphysema.