Triglyceride e test wako

Plasma TG and TC were gauged by triglyceride/GB and cholesterol/HP kits (Boehringer Mannheim, Mannheim, Germany), respectively. Liver homogenate (1 mL) was mixed with 2.5 mL chloroform/methanol (2:1, v/v), chloroform layer collected and concentrated. After mixing with 10% Triton X-100 in isopropanol, sample was assayed by Wako Triglyceride E-Test and Total Cholesterol E-Test kits (Wako Pure Chemical, Osaka, Japan).

The BAT samples were homogenized with RIPA buffer (50 mM Tris-HCl, pH 7.4, 1% NP40, 0.25% Na-deoxycholate, 150 mM NaCl, and 1 mM EDTA) and centrifuged at 18,000×g and 4°C for 10 min. The lipids in the supernatants were extracted with methanol/chloroform (1∶2), evaporated with NO₂, and dissolved in isopropanol. The triglyceride levels were determined using the Wako Triglyceride E-test (Wako Chemical, Osaka).

Blood was collected from the neck vein plexus of anesthetized mice after administration of BTS solution or water for 25 days. Blood samples were centrifuged at 2000g for 5 min at 25 °C and the supernatant was used as the serum sample. Serum glucose concentration was measured by the glucose oxidase method using a serum glucose monitor (Medisafe Mini, Termo, Tokyo, Japan). Serum insulin was measured using a mouse enzyme linked immunosorbent assay (ELISA) kit for insulin (Morinaga, Yokohama, Japan). The homeostasis model assessment-insulin resistance (HOMA-IR) value was calculated as [serum glucose (mg/dL) × serum insulin (μU/mL)]/405 [24 (link)]. Serum leptin and adiponectin were measured with mouse Leptin and Adiponectin/Acrp30 Quantikine ELISA kits (R&B Systems, Minneapolis, USA). Serum triglyceride was measured using the Wako Triglyceride E Test (Wako, Osaka). High-density lipoprotein (HDL) cholesterol and low-density lipoprotein (LDL) cholesterol were measured using HDL and LDL/VLDL-cholesterol Quantification Kits (Bio Vision, Milpitas, USA).