Shih Andy Y, Imbeault Sophie, Barakauskas Vilte, Erb Heidi, Jiang Lei, Li Ping, Murphy Timothy H
Kinsmen Laboratory, Department of Psychiatry, Brain Research Center, Vancouver, Canada.
J Biol Chem. 2005 Jun 17;280(24):22925-36. doi: 10.1074/jbc.M414635200. Epub 2005 Apr 19.
NF-E2 related factor (Nrf2) controls a pleiotropic cellular defense, where multiple antioxidant/detoxification pathways are up-regulated in unison. Although small molecule inducers of Nrf2 activity have been reported to protect neurons in vitro, whether similar pathways can be accessed in vivo is not known. We have investigated whether in vivo toxicity of the mitochondrial complex II inhibitor 3-nitropropionic acid (3-NP) can be attenuated by constitutive and inducible Nrf2 activity. The absence of Nrf2 function in Nrf2(-/-) mice resulted in 3-NP hypersensitivity that became apparent with time and increasing dose, causing motor deficits and striatal lesions on a more rapid time scale than identically treated Nrf2(+/+) and Nrf2(+/-) controls. Striatal succinate dehydrogenase activity, the target of 3-NP, was inhibited to the same extent in all genotypes by a single acute dose of 3-NP, suggesting that brain concentrations of 3-NP were similar. Dietary supplementation with the Nrf2 inducer tert-butylhydroquinone attenuated 3-NP toxicity in Nrf2(+/-) mice, but not Nrf2(-/-), confirming the Nrf2-specific action of the inducer in vivo. Increased Nrf2 activity alone was sufficient to protect animals from 3-NP toxicity because intrastriatal adenovirus-mediated Nrf2 overexpression significantly reduced lesion size compared with green fluorescent protein overexpressing controls. In cultured astrocytes, 3-NP was found to increase Nrf2 activity leading to antioxidant response element-dependent gene expression providing a potential mechanism for the increased sensitivity of Nrf2(-/-) animals to 3-NP toxicity in vivo. We conclude that Nrf2 may underlie a feedback system limiting oxidative load during chronic metabolic stress.
核因子E2相关因子(Nrf2)控制着一种多效性细胞防御机制,在此机制中,多种抗氧化/解毒途径会同时上调。尽管已有报道称Nrf2活性的小分子诱导剂在体外可保护神经元,但尚不清楚在体内是否能激活类似途径。我们研究了线粒体复合物II抑制剂3-硝基丙酸(3-NP)的体内毒性是否可通过组成型和诱导型Nrf2活性来减轻。Nrf2基因敲除(Nrf2(-/-))小鼠中Nrf2功能的缺失导致对3-NP超敏,且随着时间推移和剂量增加愈发明显,与接受相同处理的Nrf2(+/+)和Nrf2(+/-)对照相比,在更快的时间尺度上导致运动功能缺陷和纹状体损伤。单次急性剂量的3-NP对所有基因型纹状体琥珀酸脱氢酶活性(3-NP的作用靶点)的抑制程度相同,这表明脑中3-NP的浓度相似。用Nrf2诱导剂叔丁基对苯二酚进行饮食补充可减轻Nrf2(+/-)小鼠的3-NP毒性,但对Nrf2(-/-)小鼠无效,这证实了该诱导剂在体内的Nrf2特异性作用。单独增加Nrf2活性就足以保护动物免受3-NP毒性,因为与过表达绿色荧光蛋白的对照相比,纹状体内腺病毒介导的Nrf2过表达显著减小了损伤大小。在培养的星形胶质细胞中,发现3-NP可增加Nrf2活性,导致依赖抗氧化反应元件的基因表达,这为Nrf2(-/-)动物在体内对3-NP毒性敏感性增加提供了一种潜在机制。我们得出结论,Nrf2可能是慢性代谢应激期间限制氧化负荷的反馈系统的基础。