2.7. Blood Collection and Analysis
BLOOD
Blood samples were taken for analysis at four points in time: immediately before and after MAE, and 6 and 24 h after MAE. The blood was collected into 5 mL tubes (Sarstedt) that contained an anticoagulant. The serum was separated by centrifugation and aliquoted into 500 μL portions. Then, the samples were frozen in liquid nitrogen and stored at −80 °C until further analysis. To assess the baseline and altered iron (FE) levels, plasma was collected into tubes with lithium heparin at two points in time: immediately before and after MAE. It was then analyzed using an in vitro IRON 2 (Roche/Hitachi Cobas c.) system and a Cobas C analyzer 501 (Hitachi, Tokyo, Japan).Biochemical analyses of serum ferritin, myoglobin, hepcidin, IL-6, and IL-10 levels were performed using commercially available highly sensitive enzyme-linked immunosorbent assay kits from DRG International Inc. (841 Mountain Ave, Springfield, NJ 07081, USA) and a Thermo Fisher Scientific ELISA Analyzer (Thermo Fisher Scientific, Waltham, MA, USA). Total antioxidative capacity (TAC) was measured using TAC Fast Track ELISA kit no. LDN-DM P-4100 (Omnignostica Forschungs GmbH, Austria) and 8-OHdG levels were measured using the Human 8-OHdG ELISA kit (Shanghai SunRed Biological Technology Co. Ltd., No. 6497, Shanghai, China). Lactate concentration was measured in capillary blood collected from the earlobe before and immediately after MAE, using a Dr. Lange Plus LP20 biochemical analyzer.