Venous blood was drawn after 12 h of fasting. Serum total cholesterol, triglycerides, HDL-c, and glucose were obtained by commercial colorimetric-enzymatic methods. LDL-c was calculated using the Friedwald equation. Measurements were performed using Dimension RxL equipment (Siemens Healthcare Diagnostic Inc., Newark, DE, USA) with dedicated reagents. Lp(a) was obtained by immunonephelometry using dedicated reagents for BN-II equipment from Siemens Healthcare (Marburg, Hessen, Germany). Plasma total homocysteine, insulin and folic acid concentrations were measured by a chemiluminescence immunoassay on an Immulite 2000® Analyzer (Siemens Healthcare Diagnostic Inc., Newark, DE, USA).
Serum Sirt-1 concentrations were determined using an ELISA kit (Uscn Life Science, Wuhan, Hubei, China). Sirt-1 samples, before and after interventions, were analyzed in duplicate and in the same ELISA plate and using the Multiscan FC plate reader (Thermo Fisher Scientific, Waltham, MA, USA), with a coefficient of variation of 12%, according to the manufacturer’s instructions. All analyses were performed according to manufacturers’ instructions.
Serum Sirt-1 concentrations were determined using an ELISA kit (Uscn Life Science, Wuhan, Hubei, China). Sirt-1 samples, before and after interventions, were analyzed in duplicate and in the same ELISA plate and using the Multiscan FC plate reader (Thermo Fisher Scientific, Waltham, MA, USA), with a coefficient of variation of 12%, according to the manufacturer’s instructions. All analyses were performed according to manufacturers’ instructions.