Abstract
Obesity and type 2 diabetes(T2D) are the most prevalent and serious metabolic diseases affecting people worldwide. However racial and ethnic disparities seems to be a risk factor for their development. Mexico has been named as one of the largest populations with the highest prevalence of diabetes and obesity. The aim of this study was to identify novel T2D-associated proteins in Mexican patients. Blood samples were collected from 62 Mexican patients with T2D and they were grouped according to their body mass index(BMI). A panel of 10 diabetes and obesity serum markers was determined using MAGPIX. A comparative proteomics study was performed using two-dimensional difference in-gel electrophoresis(2D-DIGE) followed by mass spectrometry(LC-MS/MS). We detected 113 spots differentially accumulated, in which 64 unique proteins were identified, proteins that were involved in metabolism pathways, molecular transport, and cellular signalling. Four proteins(14-3-3, ApoH, ZAG, and OTO3) showing diabetes-related variation and also changes in relation to obesity were selected for further validation by western blotting. Our results reveal new diabetes related proteins present in the Mexican population. These could provide additional insight into the understanding of diabetes development in Mexican population and may also be useful candidate biomarkers.
Highlights
Introduction2D-Difference Gel Electrophoresis (2D-DIGE) incorporates the use of fluorescent molecules (CyDyes), or fluors, that are used to pre-label samples prior to separation by 2-DE19, being a very sensitive gel-based proteomic technique that is unique through the utilization of fluorescently labelled samples on the same gel, and the application of an internal standard for intra- and inter-gel comparisons and normalization[24]
The patients with diabetes were grouped according to their body mass index into four groups: normal weight (NW), overweight (OW), obesity I (ObI), and obesity II and III (ObII/III)
Proteins of interest were identified in both fractions, high-abundance proteins (HAP) and low-abundance proteins (LAP), we focused on 4 proteins (Fig. 4A), two of them located in a cluster that included diabetes related proteins: 14-3-3 protein beta/alpha and Zinc-alpha-2-glycoprotein (ZAG; spots, 26, 36, and 37 from LAP); one protein included in clusters with tendencies related to diabetes and obesity, Beta-2-glycoprotein or Apoliprotein H (ApoH), and Otopetrin 3 (OTOP3, spot 37 from HAP) that showed an obesity-related behaviour
Summary
2D-DIGE incorporates the use of fluorescent molecules (CyDyes), or fluors, that are used to pre-label samples prior to separation by 2-DE19, being a very sensitive gel-based proteomic technique that is unique through the utilization of fluorescently labelled samples on the same gel, and the application of an internal standard for intra- and inter-gel comparisons and normalization[24]. This technique has been used for studies of hepatocellular carcinoma[25], alterations in human endometriosis[26] and patients with diabetes and cardiovascular disease[27]. Serum biomarkers with the ability to differentiate between these phenotypes would have clinical relevance with regard to diagnosis, and to long-term prognosis
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