Observation Of Mice
Mice were placed in metabolic cages at night and starved for 12 hours before sacrifice. Thereafter, nine mice in total per experimental group were sacrificed at 13, 18 and 24 weeks. Dissecting stage was prepared and the animals were anaesthetized with isoflurane, sacrificed by cervical dislocation and mice were placed in a supine position and immobilized. However, blood samples were collected from the sinus by using the orbital bleeding technique and transferred into Eppendorf tubes containing heparin, which was used for fasting blood glucose measurement. The thoracic cavity was immediately dissected to remove the heart, excised, washed with phosphate-buffered saline solution at a PH of 7.4, rinsed, dried between folds of filter paper, weighed
and then divide into two parts. Homogenized and centrifuged for 15 min at 12000 rmp and 4⁰C to separate the supernatant. The left over homogenate was reserved at 20-⁰C for the analysis of total antioxidant capacity (T-OAC) and malondialdehyde (MDA). The remaining portion of the heart was quickly frozen in liquid nitrogen and kept in -80°C for later analysis
and then divide into two parts. Homogenized and centrifuged for 15 min at 12000 rmp and 4⁰C to separate the supernatant. The left over homogenate was reserved at 20-⁰C for the analysis of total antioxidant capacity (T-OAC) and malondialdehyde (MDA). The remaining portion of the heart was quickly frozen in liquid nitrogen and kept in -80°C for later analysis