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使用二维差异凝胶电泳和质谱分析法对多囊卵巢综合征患者的网膜脂肪组织进行蛋白质组学分析。

Proteomic analysis of human omental adipose tissue in the polycystic ovary syndrome using two-dimensional difference gel electrophoresis and mass spectrometry.

作者信息

Cortón Marta, Botella-Carretero José I, López Juan A, Camafeita Emilio, San Millán José L, Escobar-Morreale Héctor F, Peral Belén

机构信息

Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, E-28029 Madrid, Spain.

出版信息

Hum Reprod. 2008 Mar;23(3):651-61. doi: 10.1093/humrep/dem380. Epub 2007 Dec 22.

Abstract

BACKGROUND

Our aim was to study the protein expression profiles of omental adipose tissue biopsies obtained from morbidly obese women with or without polycystic ovary syndrome (PCOS) at the time of bariatric surgery to evaluate the possible involvement of visceral adiposity in the development of PCOS.

METHODS

Ten PCOS patients and nine control samples were included. We used two-dimensional difference gel electrophoresis (2D-DIGE) followed by in-gel digestion, and mass spectrometry (MS) of selected protein spots.

RESULTS

The 2D-DIGE technology allowed the analysis of approximately 1840 protein spots in the comparative study of control and patient proteomes, revealing 15 statistically significant spot changes (>2-fold, P < 0.05). Unambiguous protein identification was achieved for 9 of these 15 spots by MS. This preliminary study revealed differences in expression of proteins that may be involved in lipid and glucose metabolism, oxidative stress processes and adipocyte differentiation; they include proapolipoprotein Apo-A1, annexin V, glutathione S-transferase M3 (GSTM3), triosephosphate isomerase, peroxiredoxin 2 isoform a, actin and adipocyte plasma membrane-associated protein. The most relevant finding was an increase of GSTM3 in the omental fat of PCOS patients confirming previous studies conducted by our group.

CONCLUSIONS

Proteomic analysis of omental fat reveals differential expression of several proteins in PCOS patients and non-hyperandrogenic women presenting with morbid obesity. The application of this novel methodology adds further evidence to support the role of visceral adiposity in the pathogenesis of PCOS.

摘要

背景

我们的目的是研究在减肥手术时从患有或不患有多囊卵巢综合征(PCOS)的病态肥胖女性获取的网膜脂肪组织活检样本的蛋白质表达谱,以评估内脏肥胖在PCOS发生发展中可能的作用。

方法

纳入10例PCOS患者和9个对照样本。我们采用二维差异凝胶电泳(2D-DIGE),随后进行胶内消化以及对选定蛋白质斑点进行质谱分析(MS)。

结果

在对照和患者蛋白质组的比较研究中,2D-DIGE技术能够分析约1840个蛋白质斑点,发现15个具有统计学意义的斑点变化(>2倍,P<0.05)。通过质谱对这15个斑点中的9个实现了明确的蛋白质鉴定。这项初步研究揭示了可能参与脂质和葡萄糖代谢、氧化应激过程及脂肪细胞分化的蛋白质表达存在差异;这些蛋白质包括载脂蛋白Apo-A1前体、膜联蛋白V、谷胱甘肽S-转移酶M3(GSTM3)、磷酸丙糖异构酶、过氧化物酶2同工型a、肌动蛋白和脂肪细胞质膜相关蛋白。最相关的发现是PCOS患者网膜脂肪中GSTM3增加,这证实了我们团队之前进行的研究。

结论

网膜脂肪的蛋白质组学分析揭示了PCOS患者和患有病态肥胖的非高雄激素女性中几种蛋白质的差异表达。这种新方法的应用为支持内脏肥胖在PCOS发病机制中的作用提供了进一步证据。

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