Proteomics Platform, CIC bioGUNE, CIBERehd, ProteoRed, Technology Park of Bizkaia, Derio, Bizkaia, Spain.
Proteomics Clin Appl. 2010 Apr;4(4):362-71. doi: 10.1002/prca.200900119. Epub 2010 Feb 3.
Non-alcoholic fatty liver disease (NAFLD) is an important cause of chronic liver injury that has gained concern in clinical hepatology. The principal aim of this study was to find differences in protein expression between patients with NAFLD and healthy controls.
Changes in protein expression of liver samples from each of the three groups of subjects, controls, non-alcoholic steatosis, and non-alcoholic steatohepatitis (NASH), were analyzed by DIGE combined with MALDI TOF/TOF analysis, a proteomic approach that allows to compare hundreds of proteins simultaneously.
Forty-three proteins exhibiting significant changes (ratio ≥1.5, p<0.05) were characterized, 22 comparing steatosis samples versus control samples and 21 comparing NASH versus control samples. Ten of these proteins were further analyzed by Western blot in tissue samples to confirm the observed changes of protein expression using DIGE. The proteins validated were further tested in serum samples of different cohorts of patients.
Following this approach we identified two candidate markers, carbamoyl phosphate synthase 1 and 78 kDa glucose-regulated protein, differentially expressed between control and NASH. This proteomics approach demonstrates that DIGE combined with MALDI TOF/TOF and Western blot analysis of tissue and serum samples is a useful approach to identify candidate markers associated with NAFLD, resulting in proteins whose level of expression can be correlated to a disease state.
非酒精性脂肪性肝病（NAFLD）是一种重要的慢性肝损伤病因，在临床肝脏病学中受到关注。本研究的主要目的是寻找 NAFLD 患者与健康对照之间蛋白质表达的差异。
通过 DIGE 结合 MALDI TOF/TOF 分析，对三组受试者（对照组、非酒精性脂肪变性和非酒精性脂肪性肝炎（NASH））的肝组织样本中的蛋白质表达变化进行分析，这是一种允许同时比较数百种蛋白质的蛋白质组学方法。
鉴定出 43 种表达发生显著变化（比值≥1.5，p<0.05）的蛋白质，其中 22 种比较脂肪变性样本与对照样本，21 种比较 NASH 与对照样本。这 10 种蛋白质进一步通过 Western blot 在组织样本中进行分析，以使用 DIGE 确认观察到的蛋白质表达变化。验证的蛋白质在不同患者队列的血清样本中进一步进行了测试。
通过这种方法，我们鉴定出两种候选标志物，即氨基甲酰磷酸合成酶 1 和 78 kDa 葡萄糖调节蛋白，它们在对照和 NASH 之间的表达存在差异。这种蛋白质组学方法表明，DIGE 结合 MALDI TOF/TOF 和组织及血清样本 Western blot 分析是一种有用的方法，可识别与 NAFLD 相关的候选标志物，鉴定出其表达水平与疾病状态相关的蛋白质。
