Plasma SIRT1 and Fetuin-A Levels in Participants

At the beginning and end of the study, ten ccs of venous blood samples were taken, and after the separation of serum, SIRT and Fetuin levels were measured in the plasma of the participants. Blood samples were collected into tubes with and without EDTA. Blood samples without EDTA Centrifuged (Beckman Avanti J-25, USA) at a rate of 3000 rpm for 10 min in order to the separation of serum. Blood samples were kept at − 70 °C (Snijdes, Germany) in Diabetes Research Center and then transferred to the laboratory of Velayat Hospital of Qazvin University of Medical Sciences, and the measurements were carried out. Fasting plasma glucose (FPG) concentration was measured by the enzymatic method using an Abbot ModelAclyon 300, USA auto analyzer with Pars-Azmone kit (Tehran, Iran). Plasma insulin was measured by using a chemiluminescent immunoassay method (LIAISON analyzer (310360) Diasorin S.P.A., Verecelli, Italy). Insulin resistance (HOMA-IR) was calculated according to the following formula: HOMA-IR = (fasting insulin (U/ml) × FPG (mg/dl)/405) [17 (link)]. Serum levels of SIRT1 and Fetuin-A in participants’ plasma were measured using a special kit and by ELISA method (Diameter, Italy and Bioassay Technology Laboratory, China).