2.4. Analysis of Metabolism Biomarkers
MP, Diabetes
DIABETES
After collecting the blood samples, serum and plasma were obtained by centrifugation at 3000× C-peptide, ghrelin, gastric inhibitory polypeptide (GIP), glucagon-like peptide-1 (GLP-1), leptin, glucagon insulin, plasminogen activator inhibitor (PAI-1), resistin and visfatin concentrations were analysed in plasma using the Bio-Rad Multiplex Diabetes kit (Ref.: 171A7001M). The plasma samples were diluted 1:4 with a Bioplex sample diluent included with the kit prior to analysis. Adipsin and adiponectin concentrations were analysed using the Bio-Rad Multiplex Adipokine kit (Ref.: 171A7002M). In this case, a 1:2500 dilution with a serum-based diluent was employed according to the kit manufacturer’s instructions. In both cases, 50 µL of the diluted plasma sample was placed in each well of the assay plate in duplicate. The analytes were measured on a Bio-Plex MAGPIX™ Multiplex reader connected to a Bio-Plex ProTM Wash Station. The software Bio-Plex Manager™ MP (Luminex Corporation, Austin, TX, USA) was used for data processing. Results were expressed as pg/mL or ng/mL plasma.