Overexpression of acyl-CoA synthetase-1 increases lipid deposition in hepatic (HepG2) cells and rodent liver in vivo
| Autor: | Gregory J. Cooney, Edward W. Kraegen, Mercedes Ballesteros, Donna Wilks, Elaine Preston, Leonie Wood, Lee Carpenter, Heidi A. Parkes, Stuart M. Furler |
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| Rok vydání: | 2006 |
| Předmět: |
Male
medicine.medical_specialty Physiology G protein Endocrinology Diabetes and Metabolism Green Fluorescent Proteins Immunoblotting Fatty Acids Nonesterified Biology Adenoviridae Eating Mice chemistry.chemical_compound In vivo Cell Line Tumor Physiology (medical) Internal medicine Coenzyme A Ligases medicine Animals Obesity Rats Wistar Beta oxidation Triglycerides chemistry.chemical_classification Triglyceride Fatty acid metabolism Reverse Transcriptase Polymerase Chain Reaction Fatty acid Lipid metabolism Organ Size Lipid Metabolism Rats Mice Inbred C57BL Enzyme Endocrinology Liver chemistry RNA Oleic Acid |
| Zdroj: | American Journal of Physiology-Endocrinology and Metabolism. 291:E737-E744 |
| ISSN: | 1522-1555 0193-1849 |
| DOI: | 10.1152/ajpendo.00112.2006 |
| Popis: | Accumulation of intracellular lipid in obesity is associated with metabolic disease in many tissues including liver. Storage of fatty acid as triglyceride (TG) requires the activation of fatty acids to long-chain acyl-CoAs (LC-CoA) by the enzyme acyl-CoA synthetase (ACSL). There are five known isoforms of ACSL (ACSL1, -3, -4, -5, -6), which vary in their tissue specificity and affinity for fatty acid substrates. To investigate the role of ACSL1 in the regulation of lipid metabolism, we used adenoviral-mediated gene transfer to overexpress ACSL1 in the human hepatoma cell-line HepG2 and in liver of rodents. Infection of HepG2 cells with the adenoviral construct AdACSL1 increased ACSL activity >10-fold compared with controls after 24 h. HepG2 cells overexpressing ACSL1 had a 40% higher triglyceride (TG) content (93 ± 3 vs. 67 ± 2 nmol/mg protein in controls, P < 0.05) after 24-h exposure to 1 mM oleate. Furthermore, ACSL1 overexpression produced a 60% increase in cellular LCA-CoA content (160 ± 6 vs. 100 ± 6 nmol/g protein in controls, P < 0.05) and increased [14C]oleate incorporation into TG without significantly altering fatty acid oxidation. In mice, AdACSL1 administration increased ACSL1 mRNA and protein more than fivefold over controls at 4 days postinfection. ACSL1 overexpression caused a twofold increase in TG content in mouse liver (39 ± 4 vs. 20 ± 2 μmol/g wet wt in controls, P < 0.05), and overexpression in rat liver increased [1-14C]palmitate clearance into liver TG. These in vitro and in vivo results suggest a pivotal role for ACSL1 in regulating TG synthesis in liver. |
| Databáze: | OpenAIRE |
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