Plasma aliquots were used to measure glucose (Spinreact, Girona, Spain), NEFA (nonesterified fatty acids) (Wako, Neuss, Germany), glycerol (Sigma Chemical, St. Louis, MO), and triglycerides (Spinreact) by an enzymatic colorimetric tests using commercial kits. Insulin was determined in plasma samples using a specific ELISA kit for rats (Mercodia, Uppsala, Sweden). Leptin and adiponectin were assayed in plasma samples using a specific enzyme immunoassay (EIA) kit for rats (Biovendor, Brno, Czech Republic; and Millipore, Bedford, MA, respectively). A Chemiluminiscent Microparticle Immunoassay (CMIA, Architect) was used for the quantitative determination of estradiol in plasma (Abbott, Ireland).
Two hundred milligrams of frozen liver were homogeneized into 2.5 mL of 0.9% ClNa, and used for lipid extraction following the Bligh and Dyer [26] method. Aliquots of lipid extracts were dried and the remaining residue weighed. Ten microliters of these homogenates were used to determine triglycerides as mentioned above.
Two hundred milligrams of frozen liver were homogeneized into 2.5 mL of 0.9% ClNa, and used for lipid extraction following the Bligh and Dyer [26] method. Aliquots of lipid extracts were dried and the remaining residue weighed. Ten microliters of these homogenates were used to determine triglycerides as mentioned above.