Inhibitors of Fatty Acid Synthesis Induce PPARα-Regulated Fatty Acid β-Oxidative Genes: Synergistic Roles of L-FABP and Glucose
Cytosolic glucose (panel a) and free fatty acid (LCFA, panel b) levels in mouse primary hepatocytes cultured with and without TOFA or C75. Hepatocytes were incubated with TOFA or C75 (10 μg/mL) with 6 or 20 mM glucose (Section 2). Cytosolic glucose level was then determined as in Methods. Briefly, hepatocytes were washed quickly with ice old solution of MgCl2 (100 mM) with 0.1 mM phloretin. Cells were then homogenized with a probe sonicator and, after centrifugation, the supernatant was used for glucose analysis with the Amplite Glucose Quantitation Kit as instructed by the manufacturer. For LCFA determination, hepatocyte homogenate was extracted twice with 1% Triton X-100 in pure chloroform. The organic phase was collected, and the FFA content of each sample was measured with the Free Fatty Acid Quantification Kit from BioVision, Inc. according to manufacturer’s instructions using enclosed palmitic acid as standard. Mean ± SEM, .