Rab0904

Triglycerides and total cholesterol concentrations were determined by enzymatic colorimetric methods using available commercial kits (Wiener lab Group, Argentina). The high-density lipoprotein cholesterol (HDLc) was determined after precipitation of very low-density lipoprotein (VLDL) and low-density lipoprotein (LDLc) with polyanions (dextran sulphate and magnesium chloride). LDL-cholesterol concentration was determined by a two-step homogeneous assay without precipitation. Free fatty acids were determined using available commercial kits (ab65341, Abcam, USA).
Circulating tumor necrosis factor-α (TNF-α) (RAB0479), interleukin-1β (IL-1β) (RAB0277), insulin (RAB0904) (all from SIGMA Aldrich, St. Louis, MO, USA), leptin (ab100773), and adiponectin (ab108784) (Abcam, USA) were performed using commercially available kits according to the manufacturer′s instructions.
Blood glucose concentrations were measured using a glucose meter (Roche Diagnostics GmbH, Mannheim, Germany). The HOMA-IR (Homeostasis model assessment of insulin resistance) index was calculated as [fasting glucose (mg/dl) × fasting insulin (ng/ml)/405] to assess insulin resistance [38 (link)].

Eight weeks after arrival, all rats underwent an oral glucose tolerance test (OGTT) to confirm glucose intolerance in diabetic rats. Animals were fasted overnight and then given an oral dose of glucose solution (BioShop Canada Inc.) at 1 g/kg via gavage. Blood glucose was measured over 2 h, prior to gavage and at 30-min intervals post-glucose consumption.
Nine weeks after arrival, all rats underwent an insulin tolerance test (ITT) to assess insulin resistance in diabetic rats. Animals were fasted for 6 h and injected IP with insulin solution (0.75 U/kg, Sigma-Aldrich, I9278). Blood glucose was measured prior to injection, at 15-min intervals post-injection for the first hour, and 30-min intervals for the second hour.
One-month post-diabetes (11 weeks after arrival), blood plasma insulin levels were tested in all rats to confirm insulin resistance in diabetic rats. Animals were fasted overnight, and blood from the saphenous vein was collected in lithium-heparin tubes. Tubes were centrifuged for 8 min at 8,000 rcf and blood plasma was separated from the samples. A rat insulin ELISA kit (Sigma-Aldrich, RAB0904) was used to determine insulin levels in the plasma samples. A homeostatic model assessment of insulin resistance (HOMA-IR) was then calculated as $\frac{Insulin(u\frac{IU}{mL})xFastingGlucose(\frac{mmol}{L})}{22.5}$ to assess insulin resistance (14 (link)).