A110 1 1 for tg

Mice were fasted for 6 h, and fasting blood-glucose (FBG) levels were examined by using a glucometer (Countour TS, Bayer, Parsipanny, NJ, United States). Blood lipid profiles, including total cholesterol (TC), triglycerides (TG), high-density lipoproteins (HDL-C) and low-density lipoproteins (LDL-C), and TC and TG contents in the liver were all measured using commercial kits (A111-2-1 for TC, A110-1-1 for TG, A112-2-1 for HDL-C and A113-2-1 for LDL-C, JianCheng Bioengineering Institute, Nanjing, China). The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by AST and ALT kits (C009-2-1 and C010-2-1, respectively, JianCheng Bioengineering Institute). The concentration of TNFα in the liver was examined by ELISA (EK1352, Signalway Antibody, Nanjing, China). All of the operations were performed strictly in accordance with the manufacturer’s protocols.

LEAP2 levels of CSF, plasma, and hypothalamic tissue samples were determined by a LEAP2 (38–77) (human)/LEAP2 (37–76) (mouse) enzyme-linked immunosorbent assay (ELISA) kit (EK-075-40, Phoenix Pharmaceuticals, Inc., Burlingame, CA, USA) following the manufacturer’s instructions. The minimum detectable concentration of LEAP2 in this assay was 0.22 ng/ml. Hypothalamic LEAP2 content was normalized to the amount of protein.
Plasma total cholesterol (TC) or triglyceride (TG) levels were measured using the corresponding assay kits (A111-1-1 for TC and A110-1-1 for TG, Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s instructions.