Proteomic similarities in steroid responsiveness in normal and glaucomatous trabecular meshwork cells
Proteomic similarities in steroid responsiveness in normal and glaucomatous trabecular meshwork cells
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发表时间：
2012-07
期刊：
影响因子：
2.2
通讯作者：
K. Bollinger;J. Crabb;Xianglin Yuan;Tasneem Putliwala;A. Clark;J. Crabb
中科院分区：
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作者：
K. Bollinger;J. Crabb;Xianglin Yuan;Tasneem Putliwala;A. Clark;J. Crabb
目的糖皮质激素(GCs)是一种常见的抗炎药，可导致眼内高眼压和继发性青光眼，其原因是房水流经小梁网(TM)受损。GC信号转导机制复杂且知之甚少。为了更好地了解眼睛中的GC信号，我们测试了一种假设，即来自正常和青光眼捐赠者的TM细胞中存在类固醇反应的常见机制。方法4种原代培养的正常人和青光眼供者的TM细胞经地塞米松(Dex)或不加地塞米松(Dex)处理10d后，提取细胞蛋白，用液质联用(LC-MS/MS)iTRAQ(等压相对和绝对定量标记)技术进行鉴定和定量。结果共检测到718个蛋白质。地塞米松处理显著改变了≥3细胞样本中40种蛋白质的丰度，其中37种蛋白质以前没有与TM细胞中的GC信号相关。在分析的所有四个TM细胞中，大多数类固醇反应蛋白都发生了变化，包括正常和青光眼。GC诱导的蛋白质组学改变支持细胞外基质的重塑，细胞骨架/细胞-细胞相互作用的破坏，以及线粒体功能障碍。这样的生理后果似乎与TM细胞中由转化生长因子-β-2诱导的相同，后者是导致高眼压和青光眼病理的另一个推定因素。结论扩展了参与GC信号转导的TM蛋白质谱，揭示了GC信号转导在正常和青光眼TM细胞中的共同后果，揭示了类固醇和转化生长因子β-2诱导的正常和青光眼TM细胞蛋白质组学变化的相似性。最后，这些数据将为TM定量蛋白质组数据库做出贡献。
Purpose Glucocorticoids (GCs) are common anti-inflammatory agents that can cause ocular hypertension and secondary glaucoma as a consequence of impaired aqueous humor outflow through the trabecular meshwork (TM). Mechanisms of GC-signaling are complex and poorly understood. To better understand GC-signaling in the eye, we tested the hypothesis that common mechanisms of steroid responsiveness exist in TM cells from normal and glaucomatous donors. Methods Four primary cultures of human TM cells from normal and glaucomatous donors were treated with or without dexamethasone (Dex) for 10 days, then cellular proteins were extracted, identified and quantified by liquid chromatography tandem mass spectrometry (LC MS/MS) iTRAQ (isobaric tags for relative and absolute quantitation) technology. Results A total of 718 proteins were quantified. Dex-treatment significantly altered the abundance of 40 proteins in ≥3 cell samples, 37 of which have not previously been associated with GC-signaling in TM cells. Most steroid responsive proteins were changed in all four TM cells analyzed, both normal and glaucomatous. GC-induced proteomic changes support remodeling of the extracellular matrix, disorganization of the cytoskeleton/cell-cell interactions, and mitochondrial dysfunction. Such physiologic consequences appear common to those induced in TM cells by transforming growth factor-β2, another putative contributor to ocular hypertension and glaucoma pathology. Conclusions The results expand the repertoire of TM proteins involved in GC-signaling, demonstrate common consequences of GC-signaling in normal and glaucomatous TM cells, and reveal similarities in proteomic changes induced by steroids and TGFβ2 in normal and glaucomatous TM cells. Finally, the data contributes to a TM quantitative proteomic database.