Rat il 10 kit

Liver tissues were homogenized as described in a previous study [21 (link)]. The homogenized solution was centrifuged at 3000× g and 4 °C for 15 min. Commercial kits were used to analyze the supernatant, including the DuoSet^® rat tumor necrosis factor (TNF)-α kit, the rat interleukin (IL)-1β/IL-1F2 kit, the rat IL-6 kit, and the rat IL-10 kit (R&D Systems, Minneapolis, MN, USA). A microplate reader (Molecular Devices, Sunnyvale, CA, USA) was used to read the optical density (OD) at 450 nm for all cytokines.

Inflammatory cytokines including tumor necrosis factor- (TNF-) α, IL-1β, IL-6, and IL-10 levels were measured. Liver tissues (0.5 g) were homogenized in 1.5 mL ice-cold buffer (50 mM Tris (pH 7.2), 150 mM NaCl, and 1% Triton-X 100) plus 0.1% of a protease inhibitor. The homogenates were shaken on ice for 90 min and then the homogenates were centrifuged at 3000 ×g and 4°C for 15 min. The supernatant was analyzed with a DuoSet® rat TNF-α kit, a rat IL-1β/IL-1F2 kit, a rat IL-6 kit, and a rat IL-10 kit (R&D Systems, Minneapolis, MN, USA). Procedures followed the assay kit instructions. The optical density (OD) was read at 450 nm for all cytokines using a microplate reader (Molecular Devices, Sunnyvale, CA, USA).

(1) Cytokine Measurements. Ice-cold buffer (1.5 mL) containing 50 mM Tris (pH 7.2), 150 mM NaCl, 1% Triton-X, and 0.1% protease inhibitor was added to the liver tissue (0.5 g) and then homogenized and shaken on ice for 90 min. The homogenized solution was centrifuged at 3000 ×g and 4°C for 15 min. A DuoSet® rat TNF-α kit, a rat IL-1β/IL-1F2 kit, a rat IL-6 kit, and a rat IL-10 kit (R&D Systems, Minneapolis, MN, USA) were used to analyze the supernatant according to assay kit instructions. A microplate reader (Molecular Devices, Sunnyvale, CA, USA) was used to read the optical density (OD) at 450 nm for all cytokines.
(2) Plasma Adiponectin Concentration. An enzyme-linked immunosorbent assay (ELISA) kit (AssayMax Rat Adiponectin ELISA kit Assaypro, St. Charles, MO, USA) was used to measure the plasma adiponectin concentration. The OD was the same as for the cytokine measurements.
(3) Cell Adhesion Molecule Measurement. Plasma VCAM-1 and ICAM-1 levels were, respectively, determined with a rat ICAM-1/CD54 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA) and Cell Adhesion Molecule 1 Assay Kit (USCN Life Science, Wuhan, China). Procedures followed the manufacturer's instructions. The OD was the same as for the cytokine measurements.