Blood Sampling and Primary Measures
Venous blood samples were collected in the fasting state for biochemical measurements. Samples were centrifuged and serums were frozen and stored at -80 °C. The serum level of IL-6 was determined using the method described by the Boster human IL-6 ELISA kit. Malondialdehyde (MDA) level was measured as an index of lipid peroxidation using TBRAS assay protocol. The MDA reacts with thiobarbituric acid (TBA) as a thiobarbituric acid reactive substance (TBARS) to produce a red colored complex. The absorbance of the samples was measured at 532nm using ELISA plate reader. 3 ml phosphoric acid (1%) and 1 ml TBA (0.6%) was added to 0.5 ml of homogenate in a centrifuge tube and the mixture was heated for 45 min in a boiling water bath. After cooling, 4 ml of n-butanol was added the mixture and vortex-mixed for 1 min followed by centrifugation at 20000 rpm for 20 min. The organic layer was transferred to a fresh tube and its absorbance was measured at 532 nm 16.