title: Dopamine induces lipid accumulation, NADPH oxidase-related oxidative stress, and a proinflammatory status of the plasma membrane in H9c2 cells.
abstract: Excess catecholamine levels are suggested to be cardiotoxic and to underlie stress-induced heart failure. The cardiotoxic effects of norepinephrine and epinephrine are well recognized. However, although cardiac and circulating dopamine levels are also increased in stress cardiomyopathy patients, knowledge regarding putative toxic effects of excess dopamine levels on cardiomyocytes is scarce. We now studied the effects of elevated dopamine levels in H9c2 cardiomyoblasts. H9c2 cells were cultured and treated with dopamine (200 μM) for 6, 24, and 48 h. Subsequently, the effects on lipid accumulation, cell viability, flippase activity, reactive oxygen species (ROS) production, subcellular NADPH oxidase (NOX) protein expression, and ATP/ADP and GTP/GDP levels were analyzed. Dopamine did not result in cytotoxic effects after 6 h. However, after 24 and 48 h dopamine treatment induced a significant increase in lipid accumulation, nitrotyrosine levels, indicative of ROS production, and cell death. In addition, dopamine significantly reduced flippase activity and ATP/GTP levels, coinciding with phosphatidylserine exposure on the outer plasma membrane. Furthermore, dopamine induced a transient increase in cytoplasmic and (peri)nucleus NOX1 and NOX4 expression after 24 h that subsided after 48 h. Moreover, while dopamine induced a similar transient increase in cytoplasmic NOX2 and p47phoxexpression, in the (peri)nucleus this increased expression persisted for 48 h where it colocalized with ROS. Exposure of H9c2 cells to elevated dopamine levels induced lipid accumulation, oxidative stress, and a proinflammatory status of the plasma membrane. This can, in part, explain the inflammatory response in patients with stress-induced heart failure.
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands; mpv.begieneman@gmail.com.|Netherlands Forensic Institute, The Hague, the Netherlands.|Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, the Netherlands.
authors: Mark P V
authors: MP
authors: Begieneman
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.|Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, the Netherlands.|Interuniversity Cardiology Institute of the Netherlands, Utrecht, the Netherlands.
authors: Ellis N
authors: EN
authors: Ter Horst
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.
authors: Liza
authors: L
authors: Rijvers
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.
authors: Elisa
authors: E
authors: Meinster
authors: Laboratory Genetic Metabolic Diseases and Department of Pediatrics/Emma's Children Hospital, Academic Medical Center Amsterdam, Amsterdam, the Netherlands.
authors: René
authors: R
authors: Leen
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.
authors: Jeannette E
authors: JE
authors: Pankras
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.
authors: Jan
authors: J
authors: Fritz
authors: Netherlands Forensic Institute, The Hague, the Netherlands.|Department of Pathology, Maastricht University Medical Center, Maastricht, the Netherlands.
authors: Bela
authors: B
authors: Kubat
authors: Department of Physiology, VU University Medical Center, Amsterdam, the Netherlands.
authors: René J P
authors: RJ
authors: Musters
authors: Interuniversity Cardiology Institute of the Netherlands, Utrecht, the Netherlands.
authors: André B P
authors: AB
authors: van Kuilenburg
authors: Core Facility Cellular Imaging/LCAM-AMC, Amsterdam, the Netherlands; and.
authors: Jan
authors: J
authors: Stap
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.|Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, the Netherlands.|Department of Cardiothoracic Surgery, VU University Medical Center, Amsterdam, the Netherlands.
authors: Hans W M
authors: HW
authors: Niessen
authors: Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.|Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, the Netherlands.
authors: Paul A J
authors: PA
authors: Krijnen
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival; D004298:Dopamine / Q000494:pharmacology*; D015259:Dopamine Agents / Q000494:pharmacology*; D005434:Flow Cytometry; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007249:Inflammation / Q000378:metabolism*; D050356:Lipid Metabolism / Q000187:drug effects*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D032386:Myoblasts, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D009247:NADH, NADPH Oxidoreductases / Q000187:drug effects / Q000378:metabolism; D000074624:NADPH Oxidase 1; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000187:drug effects* / Q000378:metabolism; D009687:Nuclear Proteins / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D009195:Peroxidase / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D014443:Tyrosine / Q000031:analogs & derivatives / Q000187:drug effects / Q000378:metabolism
chemical_list: D015259:Dopamine Agents; C498751:Nsfl1c protein, rat; D009687:Nuclear Proteins; D017382:Reactive Oxygen Species; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D006153:Guanosine Diphosphate; C002744:3-nitrotyrosine; D014443:Tyrosine; D000244:Adenosine Diphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D009195:Peroxidase; D009247:NADH, NADPH Oxidoreductases; D000074624:NADPH Oxidase 1; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C000614712:NOX1 protein, rat; C498911:Nox4 protein, rat; C505410:Casp3 protein, rat; D053148:Caspase 3; D004298:Dopamine
keywords: reduced nicotinamide adenine dinucleotide phosphatase oxidase proteins