Bs 220

Serum levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein-cholesterol (HDL), and low-density lipoprotein-cholesterol (LDL), as well as aspartate aminotransferase (AST) and alanine aminotransferase (ALT), were measured Automated Biochemistry Analyzer (BS220, Mindray, China). Additionally, commercial enzyme-linked immunosorbent assay (ELISA) kits were used to measure serum levels of leptin (Abcam, Cambridge, UK) and adiponectin (R&D systems, Minneapolis, MN, USA).

Orbital venous plexus blood and white fat were collected from mice after the 8-week treatment; blood serum was separated. Serum triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels were measured using an auto-biochemical analyzer (Mindray BS-220, China). Fasting blood glucose (FBG) levels were measured using a blood glucose meter. Fasting insulin (FIN), gastric inhibitory peptide (GIP), and glucagon-like peptide-1 (GLP-1) levels were determined using Bio-Plex suspension chip system (Bio-Plex 200 System) with Bio-Plex Pro™ Assay kit, and the insulin resistance index (HOMA-IR) and insulin sensitivity index (ISI) were calculated. Following the manufacturer’s protocol, glycogen content in the liver and muscle was measured using respective assay kits. Similarly, glucose consumption and cellular glycogen content were measured using assay kits. HOMA-IR index = FBG (mM) × FIN (mIU/L)/22.5; ISI = 1/[FBG (mM) × FIN (mIU/L)]; white fat index (WFI) = white fat (mg)/body weight of mouse (g).

Blood samples were collected at the Hainan Provincial People’s Hospital for the measurements of clinical indices, including metabolic parameters (TGs, TC, and FPG), sex hormones (LH, FSH, LH/FSH, E2, PRL, and T) and gut-brain mediators (PYY and ghrelin). The sex hormones were measured on day 3 of the menstrual cycle. The blood biochemical indices were measured on an automatic biochemical analyzer (BS-220; Mindray, China), and the sex hormones were tested using an automated immunoassay system (AIA-1200; Tosoh, Japan). The ghrelin and PYY levels were determined using commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions (12 (link)). To determine SCFA levels, we used approximately 250 mg of frozen fecal samples. The concentrations of SCFAs were determined using a 1:25 dilution of 500 μl of supernatant. Gas chromatography-mass spectrometry (GC-MS) determinations were performed using a Varian Saturn 2000 GC-MS instrument with an 8200 CX solid-phase microextraction (SPME) autosampler (54 (link)).

Lactate dehydrogenase (LDH) and creatinine kinase (CK) were measured using a blood biochemical analyzer (BS 220, Mindray, Shenzhen, China).

Mouse and rat serum ALT and AST levels were detected using a Fully Automatic Biochemical Analyzer (BS‐220, Mindray).

Venous blood samples (20 mL) were drawn from the antecubital vein in the morning after 12 h of overnight fasting. Serum was separated within 30 min and stored at −80 °C until analyses. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and triglycerides were analyzed using the enzymatic, colorimetric method with an automatic biochemical analyzer (Mindray BS-220, Shenzhen, China).