Mitochondrial Function and Fatty Acid Oxidation

Left ventricular samples were used for mitochondria extraction from tissue homogenates and measurement of oxidative phosphorylation (OXPHOS) (Buondonno et al., 2016 (link)) and ATP concentration were performed as previously described (Campia et al., 2009 (link)). The rate of reduction of cytochrome c, an index of the electron transport between Complex I–III and OXPHOS rate, was measured spectrophotometrically on 10 μL of non-sonicated mitochondrial samples. The results were expressed as nmoles cyt c reduced/min/mg mitochondrial proteins. FAO analysis was performed as previously reported (Capello et al., 2016 (link)), by radiolabeling for 2 h tissue homogenates with 2 μCi [1−¹⁴C] palmitic acid (3.3 mCi/mmol, PerkinElmer, Waltham, MA, United States) and measuring the labeled acid soluble metabolites (ASM), as products of FAO, by liquid scintillation. The results were expressed as pmoles ¹⁴C-ASM/h/mg proteins.