Oxiselect tbars

Porcine kidneys were obtained as described above (n = 4) and preserved on ice (4 °C) in UW solution for 16 h. Lipid peroxidation was quantified by measurement of malondialdehyde (MDA) using the OxiSelect TBARS assay kit (Cell Biolabs, USA) in samples taken 30 min after slaughter (37 °C) and after 16 h of cold storage. Lipid peroxidation levels were corrected for the amount of protein and expressed as relative to normothermic levels.

Blood samples were centrifuged at 3,000 rpm and 4°C for 15 min; serum samples were collected and stored at −70°C until analysis. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total cholesterol (T-Chol), triglyceride (TG), low-density-lipoprotein cholesterol (LDL-C), and high-density-lipoprotein cholesterol (HDL-C) were measured using an automatic blood chemistry analyzer (BS-220, Mindray, Xuzhou City, China). Measurements of osteocalcin, C-telopeptide of type I collagen (CTX-1), superoxide dismutase (SOD), malonyl dehydrogenase (MDA), endothelin-1, and nitric oxide in serum were performed using the Rat Osteocalcin ELISA kit (Cusabio Biotech, Houston, TX, USA), RatLaps (CTX-1) EIA Assay (Immunodiagnostic System, Gaithersburg, MD, USA), OxiSelect™ Superoxide Dismutase Activity (Cell Biolabs, Inc., San Diego, CA, USA), OxiSelect™ TBARS (Thiobarbituric Acid Reactive Substances) Assay Kit (MDA Quantitation, Cell Biolabs Inc., San Diego, CA, USA), Quantikine ELISA Endothelin-1 kit (R&D Systems, Minneapolis, USA), and colorimetric NO assay kit (R&D Systems, Minneapolis, MN, USA), following the manufacturer's instructions. All samples were analyzed in duplicate in the assay.