Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Reprod Toxicol. 2012 Dec;34(4):538-44. doi: 10.1016/j.reprotox.2012.08.008. Epub 2012 Sep 5.
To quantitate alcohol-induced alterations in the maternal uterine endothelial proteome utilizing iTRAQ-based mass spectrometry.
Uterine artery endothelial cells from third trimester pregnant ewes were FAC sorted, validated and treated without or with binge-like alcohol. Lysates were trypsin digested, iTRAQ-labeled, and analyzed using nano LC MS/MS.
Alcohol significantly upregulated 14 and downregulated 17 proteins (P<0.05) including those related to cell structure, transcription/translation regulation, histones, Ca(2+)/NO, and redox balance. Gene Ontology and ArrayTrack analyses revealed alterations to protein processing, binding, and nutrient metabolism pathways. Further, alcohol altered proteins previously correlated with fetal alcohol spectrum disorders (FASD) and those that regulate epigenetic, transcriptional, and translational processes.
Alcohol differentially alters the proteome in the maternal uterine compartment at the level of the endothelium. iTRAQ mass spectrometry provides a robust high throughput platform to comprehend the multi-mechanistic actions of alcohol and develop appropriate biomarkers and ameliorative measures for FASD.
利用 iTRAQ 基于质谱的方法定量酒精对母体子宫内皮蛋白质组的影响。
从妊娠晚期母羊的子宫动脉内皮细胞中进行 FAC 分选、验证，并进行无或 binge-like 酒精处理。用胰蛋白酶消化裂解物，用 iTRAQ 标记，并使用纳升 LC-MS/MS 进行分析。
酒精显著上调了 14 种蛋白，下调了 17 种蛋白（P<0.05），包括与细胞结构、转录/翻译调控、组蛋白、Ca(2+)/NO 和氧化还原平衡相关的蛋白。GO 和 ArrayTrack 分析揭示了蛋白质加工、结合和营养代谢途径的改变。此外，酒精改变了与胎儿酒精谱系障碍（FASD）相关的蛋白，以及调节表观遗传、转录和翻译过程的蛋白。
酒精在母体子宫内皮水平上差异地改变了蛋白质组。iTRAQ 质谱提供了一个强大的高通量平台，以理解酒精的多机制作用，并为 FASD 开发适当的生物标志物和改善措施。
