Automated hematology analyzer

All mice were euthanized using pentobarbital sodium after being fasted for 8 h; fasting 1 ml blood samples were obtained by subclavian venous puncture. Serum was obtained by centrifugation of blood at 3000 rpm for 15 min. Alanine aminotransferase (ALT) was measured as an indication of liver function by an automated hematology analyzer (Bayer, PA, USA). Serum glucose and lipid parameters including total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG) were measured using Olympus AU5400 automatic analyzer in Shandong Provincial Hospital. Serum insulin levels were tested by mouse insulin ELISA Kit (CSB-E05071m).

Serum total bilirubin, direct bilirubin, and indirect bilirubin levels were measured by the vanadate oxidation method using automatic biochemical analyzer. Hematologic test was measured using an automated hematology analyzer (Bayer Diagnostics, Newbury, and Berkshire, UK). During blood routine test [red blood cell (RBC) count, white blood cell (WBC) count, platelet count, hemoglobin, mean corpuscular volume and red cell distribution width (RDW)], fasting blood glucose, and creatinine, as well as the fasting serum lipid status including levels of total cholesterol (TC), LDL-cholesterol, high-density lipoprotein (HDL)- cholesterol, triglyceride (TG), and C-reactive protein (CRP) were recorded. Height and weight were measured, and the body mass index (BMI) was calculated as weight (kg) divided by height squared (m2). The eGFR was calculated as follows [17 (link)]: 186×SCr-1.154×age in years-0.203×1.210 (if black) ×0.742(if female) 36. Then CKD was defined as eGFR <60 ml/min/1.73 m².