Accu chek active

The third day before the end of the animal experiment, 12 hours fasting adapted rats were treated with sterilized glucose solution (2 g/kg, Sigma, USA) by oral gavage. The blood glucose values were measured using ACCU-CHEK Active (Roche Diagnostics, Germany) via the tail needle-punched blood drops at 0, 30, 60, 90, 120 min after glucose treatment. The second day before the end of animal experiment, 12 hours fasting-adapted rats were administrated biosynthetic human insulin (0.75 U/kg, Eli Lilly and Company, IN, USA) by intraperitoneal injection. Blood glucose values were determined by ACCU-CHEK Active (Roche Diagnostics) via the tail needle-punched blood drops at 0, 30, 60, 90, 120 min after insulin injection. Both of the OGTT and IPITT results were expressed as areas under the curves (AUC) to evaluate the degree of the glucose tolerance impairment and insulin sensitivity separately.

Intraperitoneal glucose tolerance test (IGTT) and insulin tolerance test (ITT) were conducted on 12 h‐fasting mice on the last day of the study period. Tail vein blood was used to determine blood glucose levels using a Roche glucometer (Accu‐Chek Active, Roche, Mannheim, Germany) at time intervals of t = 15, 30, 45, 60, 90, and 120 min after intraperitoneal injection of 0.5 U insulin/kg (Sigma) and 2 g glucose/kg (Sinopharm Chemical Reagent Co. Ltd.). Homeostatic model assessment of insulin resistance (HOMA‐IR = fasting blood glucose [mmol/L] × insulin [mIU/L]/22.5) was used to estimate insulin resistance (Matthews et al., 1985 (link)). At the end of the experimental period, the mice were anesthetized with 20% urethane and blood samples were obtained via cardiac puncture. Serum samples were collected by centrifugation at 3000 × g for 15 min. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triacylglycerol (TG), glucose, insulin, tumor necrosis factor‐α (TNF‐α), interleukin‐2 (IL‐2), lipopolysaccharide (LPS), and liver TG and TC were measured using commercial ELISA kits from Sino Best Biological Technology Co. Ltd.

An insulin tolerance test (ITT) was performed in GK rats (n = 5 for each group) 14 days after being transfected with 5×10⁸ TU of lentiviral vectors with or without the human ApoM gene via tail vein injection. After a 5-h fast, rats were injected with 1 IU/kg of insulin (Wanbang Biopharmaceuticals, China) intra-peritoneally. Blood was sampled from the tail vein 0, 30, 60, 90, 120, 150, 180 and 210 min after insulin injection. Blood glucose levels were determined using a glucosimeter (ACCU-CHEK Active, Roche Diagnostics, Basel, Switzerland). Blood glucose half-time (t1/2) was calculated from the slope of the least squares regression line of the blood glucose concentration during the linear phase of decrease.
After ITT, GK rats were anesthesized using 10% chloral hydrate (4 ml/kg) and sacrificed. Lungs, livers and kidneys were removed, sectioned, and stored in liquid nitrogen.