Spotchem automatic dry chemistry system

Blood samples for biochemical measurements were collected from each animal prior to and at the end of the experiment. Samples were obtained from the tail vein, collected into tubes containing ethylenediaminetetraacetic acid or heparin, and plasma was separated by centrifugation at 1000× g for10 min at 4 °C. Plasma samples were stored at −80 °C until further use. Plasma total blood urea nitrogen (BUN), creatinine, alanine transaminase (ALT), aspartate aminotransferase (AST), blood sugar, and lactic dehydrogenase (LDH) were measured using a SPOTCHEMTM automatic dry chemistry system (SP-4410; Arkray, Shanghai, China). Enzyme-linked immunosorbent assays (ELISAs) were performed to determine the plasma levels of C-reactive protein (CRP; Abcam Inc., Cambridge, MA, USA), high mobility group box 1 (HMGB1; LifeSpan Biosciences, Inc., Seattle, WA, USA), transforming growth factor β1 (TGF-β1; Abcam Inc., Cambridge, MA, USA), SDF-1α (R&D Systems Inc., Minneapolis, MN, USA), interferon-γ (INF-γ; Abcam Inc., Cambridge, MA, USA), and interleukin-2 (IL-2; Abcam Inc., Cambridge, MA, USA).

Blood samples for biochemical measurements were collected from each animal before and at the end of the experiment. Samples were obtained from the mandibular artery without sedation, collected into EDTA-containing tubes and separated by centrifugation at 1000xg for 10 min at 4°C. Plasma samples were stored at -80°C until tested. Plasma total cholesterol, triglyceride, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were measured using a SPOTCHEMTM automatic dry chemistry system (SP-4410; Arkray, Shanghai, Japan). ELISAs were performed to determine the plasma level of SDF-1α (R&D Systems Inc., Minneapolis, MN, USA), DPP-4 (RayBiotech Inc., Norcross, GA, USA), DPP-4 activity (Enzo Life Sciences, Lausen, Switzerland), and GLP-1 activity (Millipore, Billerica, MA, USA).