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.