Lf 90 instrument

BW and food intake were measured every week through the study. Blood glucose levels were measured biweekly using a glucometer (Kroger, Cincinnati, OH, USA). Body composition of the mice was examined at 0, 4, and 8 weeks after treatment using an LF-90 instrument (Bruker Optics, Inc., Billerica, MA, USA). Four and 6 weeks after treatment with kaempferol, mice fasted for 15 h were administered via i.p., injection a single dose of pyruvate (1 g/kg BW), or glucose (1 g/kg BW) for pyruvate and glucose tolerance tests, respectively. Blood glucose levels were then measured at 0, 30, 60, 90, and 180 min and 0, 15, 30, 60, and 120 min after administration of pyruvate or glucose. The area under the curve (AUC) for these tests was calculated using the trapezoidal rule [70 (link)]. At the end of the study, the mice were fasted for 15 h and were then euthanized between 9–11 a.m. Blood was collected immediately, and various organs were weighed and snap-frozen in liquid nitrogen and then stored at −80 °C for further analyses. Plasma lipid profile was analyzed by enzymatic methods using assay kits (Teco Diagnostics, Anaheim, CA, USA). Plasma insulin levels were measured using an ultrasensitive mouse insulin ELISA kit (Mercodia, Inc., Uppsala, Sweden). Plasma glucagon levels were measured using mouse glucagon ELISA kit (Crystal Chem, Downers Grove, IL, USA).