L type wako cho h

At sacrifice, blood samples were collected to measure the serum concentrations of total cholesterol, TG, very-low-density lipoprotein (VLDL), LDL, high-density lipoprotein (HDL), glucose, and adiponectin after overnight fasting from 10 or 14 mice in each group. Whole blood anti-coagulated with heparin lithium was taken from the inferior vena cava with a sterile syringe (Terumo, Tokyo, Japan). The serum was obtained by centrifugation (3000 rpm for 10 min), and stored at −80 °C until measurement. The serum TG and total cholesterol levels were determined using commercial enzymatic assay kits (TG, L-Type WAKO-TG·H; and total cholesterol, L-Type WAKO-CHO·H), obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). The serum levels of HDL, LDL, and VLDL were determined using an HDL and LDL/VLDL Cholesterol Quantitation Kit (BioVision, Inc., Milpitas, CA, USA). The serum glucose level was determined using commercial enzymatic assay kit (Glucose CII-test WAKO, Wako Pure Chemical Industries). These measurements were expressed as mg/dL. The serum adiponectin level (µg/mL) was determined with Mouse/Rat Adiponectin ELISA kits (Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan).

After 16 h of starvation, blood was collected from mice, incubated for 1 h at room temperature, and then centrifuged (1,500 g, room temperature, 15 min) to obtain serum. Serum total cholesterol (T-CHO), triglyceride (TG), non-esterified fatty acids (NEFA), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), glucose concentrations, insulin, and adiponectin were determined using L-type Wako CHO-H (Wako Pure Chemical Industries, Osaka, Japan), L-type Wako TG-H (Wako Pure Chemical Industries, Osaka, Japan), NEFA-SS Eiken (Eiken Chemical, Tochigi, Japan), Cholestest LDL (SEKISUI MEDICAL, Tokyo, Japan), Cholestest N HDL (SEKISUI MEDICAL), Quick Auto Neo GLU-HK (Sino Test, Tokyo, Japan), Ultra Sensitive Mouse Insulin ELISA kit (Morinaga Institute of Biological Science, Kanagawa, Japan) and Mouse/Rat adiponectin ELISA kit (Otsuka Pharmaceutical, Tokyo, Japan) respectively.