2.13. Serum and liver tissue homogenate preparations

By using a heparinized syringe, blood samples (approximately 4 mL) were drained out from the inferior vena cava of rats in each group. Then, the blood samples were separated into 2 portions: one portion conveying 1 mL of blood which was placed into EDTA tubes for hematological analysis, while the remaining portion was placed into plain tubes at room temperature for 30 min before centrifugation at 3000 rpm for 10 min to yield the serum needed for subsequent biochemical analysis. At the same time, liver tissues were flensed immediately from the surrounding tissues and washed with ice-cold phosphate buffer saline, followed by weighing. Subsequently, Liver samples were homogenized with phosphate buffer saline (25 mM, pH 7.4) to produce an approximately 10 % (w/v) homogenate. Centrifugation was then done at 1700 rpm for 10 min, and the supernatant was collected prior to storage at −20 °C until biochemical analysis. For histopathological examination, a portion of the liver tissues was stored in 10 % formalin and the relative organ weight gain of the liver was calculated by dividing the liver weight by the final body weight of each rat according to the following formula: relative organ weight (%) = (wet organ weight/body weight) × 100 [25].