Skin fibroblast model to study an impaired glutathione synthesis: Consequences of a genetic polymorphism on the proteome
Skin fibroblast model to study an impaired glutathione synthesis: Consequences of a genetic polymorphism on the proteome
复制标题
DOI：
10.1016/j.brainresbull.2008.10.015
复制
发表时间：
2009-04-06
影响因子：
3.8
通讯作者：
Riederer, Beat M.
中科院分区：
文献类型：
--
作者：
Gysin, Rene;Riederer, Irene M.;Riederer, Beat M.
谷胱甘肽（GSH）的合成受损，观察到在几个多因素的疾病，包括精神分裂症和心肌梗死。遗传研究揭示了精神分裂症与谷氨酸半胱氨酸连接酶（GCL）催化亚基（GCLC）中的GAG三核苷酸重复（TNR）多态性之间的关联。疾病相关的基因型，这种多态性与GCLC蛋白表达，GCL活性和GSH含量的下降。为了阐明蛋白质组水平GCL活性降低的后果，基于GCLC GAG TNR多态性选择了三名精神分裂症患者和三名对照。通过皮肤活检获得成纤维细胞培养物，并用已知诱导氧化应激的物质叔丁基对苯二酚（t-BHQ）进行激发。蛋白质组的变化进行了分析，通过二维凝胶电泳（2-DE），结果显示10个点，上调后的t-BHQ治疗的患者，但在对照组。通过MALDI质谱可以鉴定出9个相应的蛋白质，这些蛋白质参与各种细胞功能，包括能量代谢、氧化应激反应和细胞骨架重组。总之，GSH合成受损的受试者的皮肤成纤维细胞显示出对氧化应激反应的改变的蛋白质组反应。此外，该研究证实了使用成纤维细胞作为研究精神疾病易感因素的额外手段。(C)2008年爱思唯尔公司All rights reserved.
An impaired glutathione (GSH) synthesis was observed in several multifactorial diseases, including schizophrenia and myocardial infarction. Genetic studies revealed an association between schizophrenia and a GAG trinucleotide repeat (TNR) polymorphism in the catalytic subunit (GCLC) of the glutamate cysteine ligase (GCL). Disease-associated genotypes of this polymorphism correlated with a decrease in GCLC protein expression, GCL activity and GSH content. To clarify consequences of a decreased GCL activity at the proteome level, three schizophrenia patients and three controls have been selected based on the GCLC GAG TNR polymorphism. Fibroblast cultures were obtained by skin biopsy and were challenged with tert-butylhydroquinone (t-BHQ), a substance known to induce oxidative stress. Proteome changes were analyzed by two dimensional gel electrophoresis (2-DE) and results revealed 10 spots that were upregulated in patients following t-BHQ treatment, but not in controls. Nine corresponding proteins could be identified by MALDI mass spectrometry and these proteins are involved in various cellular functions, including energy metabolism, oxidative stress response, and cytoskeletal reorganization. In conclusion, skin fibroblasts of subjects with an impaired GSH synthesis showed an altered proteome reaction in response to oxidative stress. Furthermore, the study corroborates the use of fibroblasts as an additional mean to study vulnerability factors of psychiatric diseases. (C) 2008 Elsevier Inc. All rights reserved.