Metabolic Profiling in Murine Diabetes

All mice were weighed once weekly. Urine was obtained at baseline, weeks 9–10, and 13–14 in all groups using a metabolic cage. Albuminuria levels were measured by an Albumin Mouse ELISA Kit (Abcam, ab207620, Cambridge, MA, USA) and were standardized to urinary creatinine, assessed by a colorimetric assay Creatinine K^® 96–300 (Labtest, Vista Alegre, Brazil) using a biochemical analyzer Cobas Mira Plus (Roche, Basel, Switzerland). The results were reported as urinary albumin-to-creatinine ratio (UACR; g/mg). Blood glucose was quantified with a glucometer (Accu-Check Advantage Blood Glucose Monitor^® (Roche Diagnostic Corporation, Indianapolis, IN, USA) after six hours of fasting. All procedures were performed on animals at the beginning (9–11 weeks-old) and at the end of the protocol (14–15 weeks-old). Glycosuria was measured by the Glucose Liquiform^® 133-2/500 LabTest kit (Lagoa Santa, MG, Brazil), and the absorbance values were obtained by the ChemWell-T spectrophotometer LabTest (Lagoa Santa, MG, Brazil). Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) and HOMA-β, as previously described [67 (link)].