ROS production and mitochondrial dysfunction driven by PU.1-regulated NOX4-p22(phox) activation in Aβ-induced retinal pigment epithelial cell injury.
ROS production and mitochondrial dysfunction driven by PU.1-regulated NOX4-p22(phox) activation in Aβ-induced retinal pigment epithelial cell injury.
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Aβ诱导的视网膜色素上皮细胞损伤中 PU.1 调节的 NOX4-p22(phox) 激活驱动的 ROS 产生和线粒体功能障碍
作者：
Sun J;Chen J;Li T;Huang P;Li J;Shen M;Gao M;Sun Y;Liang J;Li X;Wang Y;Xiao Y;Shi X;Hu Y;Feng J;Jia H;Liu T;Sun X
基本原理：β淀粉样蛋白（Aβ）沉积是年龄相关性黄斑变性（AMD）的重要病理过程，主要由氧化应激引起视网膜色素上皮（RPE）变性。然而，尽管进行了大量的研究，但过氧化在Aβ介导的RPE损伤中的作用程度及其潜在机制尚未完全阐明。研究方法：我们对Aβ1-40诱导的视网膜变性小鼠模型中的RPE-脉络膜复合物进行了串联质量标记（TMT）质谱（MS）和生物信息学分析，以获得全面的蛋白质组学图谱。通过活性氧检测、线粒体活性氧测定、耗氧率测定、基因敲除实验、染色质免疫沉淀（ChIP）和荧光素酶测定证实了该模型中的关键调节子。结果如下：共鉴定了4243种蛋白质，其中1069种蛋白质受Aβ1-40显著影响，并通过生物信息学分析发现在氧化相关途径中富集。此外，NADPH氧化酶被鉴定为Aβ1-40介导的氧化应激中的枢纽蛋白，如线粒体功能障碍和活性氧物质过量产生所证明。通过基序和结合位点分析，我们发现转录因子PU.1/Spi 1是NADPH氧化酶，特别是NOX 4-p22 phox复合物激活的主要调节因子。此外，PU.1沉默阻止了RPE氧化应激和线粒体功能障碍，并挽救了视网膜结构和功能。结论：我们的研究表明，PU.1是AMD的一个新的治疗靶点，调节PU.1的表达代表了一种潜在的新方法，可以对抗Aβ驱动的RPE损伤中的过度氧化应激。
Rationale: Amyloid β (Aβ) deposition, an essential pathological process in age-related macular degeneration (AMD), causes retinal pigment epithelium (RPE) degeneration driven mostly by oxidative stress. However, despite intense investigations, the extent to which overoxidation contributes to Aβ-mediated RPE damage and its potential mechanism has not been fully elucidated. Methods: We performed tandem mass-tagged (TMT) mass spectrometry (MS) and bioinformatic analysis of the RPE-choroid complex in an Aβ1-40-induced mouse model of retinal degeneration to obtain a comprehensive proteomic profile. Key regulators in this model were confirmed by reactive oxygen species (ROS) detection, mitochondrial ROS assay, oxygen consumption rate (OCR) measurement, gene knockout experiment, chromatin immunoprecipitation (ChIP), and luciferase assay. Results: A total of 4243 proteins were identified, 1069 of which were significantly affected by Aβ1-40 and found to be enriched in oxidation-related pathways by bioinformatic analysis. Moreover, NADPH oxidases were identified as hub proteins in Aβ1-40-mediated oxidative stress, as evidenced by mitochondrial dysfunction and reactive oxygen species overproduction. By motif and binding site analyses, we found that the transcription factor PU.1/Spi1 acted as a master regulator of the activation of NADPH oxidases, especially the NOX4-p22phox complex. Also, PU.1 silencing impeded RPE oxidative stress and mitochondrial dysfunction and rescued the retinal structure and function. Conclusion: Our study suggests that PU.1 is a novel therapeutic target for AMD, and the regulation of PU.1 expression represents a potentially novel approach against excessive oxidative stress in Aβ-driven RPE injury.
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DOI：
10.1093/bioinformatics/btr064
发表时间：
2011-04-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Grant CE;Bailey TL;Noble WS
通讯作者：
Noble WS
硅蛋白诱导线粒体NOX4介导的内质网应激反应及其随后的凋亡。
影响因子：
3.8
作者：
Kim SH;Kim KY;Yu SN;Seo YK;Chun SS;Yu HS;Ahn SC
通讯作者：
Ahn SC
WGCNA：用于加权相关网络分析的 R 包。
影响因子：
3
作者：
Langfelder P;Horvath S
通讯作者：
Horvath S
影响因子：
11.4
作者：
Ferrington DA;Ebeling MC;Kapphahn RJ;Terluk MR;Fisher CR;Polanco JR;Roehrich H;Leary MM;Geng Z;Dutton JR;Montezuma SR
通讯作者：
Montezuma SR
影响因子：
15.9
作者：
Fan, Lampson M.;Geng, Li;Li, Jian-Mei
通讯作者：
Li, Jian-Mei