Plasma analysis
At the end of the study, fasted mice for eight hours and injected of
ketamine/xylazine (i.p. ). After the mice were fully anesthetized,
collected the whole blood from abdominal aorta in a tube containing 1 mM
EDTA for the measurement of relevant plasma parameters. Killed the mice
by cervical decapitation. Weighed the tissues of interest and
freeze-clamped or fixed them in a 4% formaldehyde solution. The whole
blood was centrifuged at a speed of 3,000 rpm for 30 minutes, and the
supernatant was separated and applied to the determination of
cholesterol, low-density lipoprotein cholesterol (LDL-c), high-density
lipoprotein cholesterol (HDL-c), and triglyceride by using correspondent
kits (Nanjing Jiancheng Bioengineering Institute, China), or detection
of glycosylated hemoglobin (HbA1c) and insulin by the ELISA detection
kit (Abcam, USA).