Chemical analyzer

Blood samples were centrifuged at 3,000× g for 10 min, and serum was separated to evaluate serum endotoxin (lipopolysaccharide, LPS) levels using a commercial kit based on the guidelines provided by the supplier (LAL, Limulus amebocyte lysate assay; Chinese Horseshoe Crab Reagent Manufactory Co., Ltd. Xiamen, China). The pro-inflammatory cytokine levels including tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin 1β (IL-1β), and anti-inflammatory cytokines such as interleukin 4 (IL-4) and interleukin 10 (IL-10) in the serum were determined using a mouse ELISA kit (Shang Hai Lengton Bioscience Co., Ltd.) Shanghai, as per manufacturer’s instructions. Alanine transaminase (ALT), aspartate transaminase (AST), total cholesterol (TC), and triglyceride (TG) levels were estimated using a chemical analyzer (Hitachi, Tokyo, Japan). Serum free fatty acids (FFAs) were measured using a commercial kit (Biovision, USA).

Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations were measured automatically using a chemical analyzer (Hitachi, Tokyo, Japan). Alkaline phosphatase (ALP) levels were measured using an ALP assay kit (Abcam, MA, USA) according to the manufacturer’s protocol [22 (link)].