Determination kit

After fasting, tail incision was made and blood (100 μL) was collected into EDTA-coated tubes. Sample was spun at 1,000 g for 15 min and plasma was aliquoted and stored at −80°C until use. Plasmatic adiponectin, leptin, TNF-α, IL-6 and MCP1 were quantified with multiplex immunoassay kits (Millipore, Molsheim, France). True triglycerides were measured with the determination kit from Sigma Aldrich (St Quentin Fallavier, France), according to manufacturer’s instructions.

The measurement of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), glutathione S-transferase (GSH S-T, EC 2.5.1.18), glutathione peroxidase (GSH-Px, EC 1.11.1), and glutathione reductase (GSH-R, EC 1.6.4.2) were performed with the Sigma-Aldrich Determination Kit (SOD, Cat. 19160, Sigma, St. Louis, MO, USA) and Cayman Kits (GSH S-T, 703302; GSH-Px, 703102; GSH-R, 703202, Cayman Chemical, Ann Arbor, MI, USA), following the manufacturer’s instructions. The catalytical activity of catalase (EC 1.11.1.6) was determined according to Aebi method (1984) [21 (link)], by the spectrometric analysis of the H₂O₂ consumption at 240 nm. Results are expressed as enzymatic activity units (U) in relation to the total protein of the samples.
The total protein content in liver (μg/mL) was determined using the Bio-Rad protein assay (Bio-Rad Laboratories, Hercules, CA, USA) based on the Bradford dye-binding method.