Glucose and Insulin Tolerance Tests

For GTTs, randomized male mice (n=6) were fasted for 16 h and then injected intraperitoneally with 2 g kg⁻¹ body weight glucose. Blood glucose values were determined by an automatic glucose monitor (Glucotrend 2, Roche) and Accu-Chek active bands (Roche)56 (link). The ITT was performed on mice fasted for 4 h, human insulin was administered by IP injection at 1 U kg⁻¹. Blood glucose levels were monitored as above immediately before injection (T0), and at 30, 60 and 120 min after injection. Blood was collected by retro-orbital puncture, centrifuged at 5,000g for 10 min and plasmas were used for the measurement of fasting blood glucose, cholesterol, triglycerides, non-essential fatty acids levels and markers of hepatic function by an Olympus AU400 chemistry analyzer.