On the mechanism of the losartan-mediated inhibition of phosphatidylcholine biosynthesis in H9c2 cells.
Phosphatidylcholine is the major phospholipid in mammalian tissues and the biosynthesis of phosphatidylcholine in H9c2 cells was previously shown to be stimulated by angiotensin II. In this study, we used the potent AT1 receptor antagonist, losartan, to determine if the angiotensin II-mediated stimulation of phosphatidylcholine biosynthesis was mediated by AT1 receptors. H9c2 cells were incubated with angiotensin II in the absence or presence of various concentrations of losartan. The cells were then incubated with [methyl-3H]choline for an additional 60 min and the radioactivity incorporated into phosphatidylcholine and its choline-containing metabolites determined. Losartan at concentrations which block AT1 receptors did not effect phosphatidylcholine biosynthesis mediated by angiotensin II. In contrast, higher concentrations of losartan inhibited radioactivity incorporated into phosphatidylcholine and its metabolites and this was due to a losartan-mediated reduction in choline uptake. Kinetic studies revealed that the losartan-mediated inhibition of choline uptake was competitive. High concentrations of losartan caused a translocation of CTP:phosphocholine cytidylyltransferase from the cytosolic (inactive) to the membrane (active) fraction likely as a compensatory mechanism for the losartan-mediated reduction in new phosphatidylcholine biosynthesis. Incubation of cells with PD123319, a potent AT2-receptor antagonist, did not block the angiotensin II-mediated stimulation of phosphatidylcholine biosynthesis. The results suggest that angiotensin II stimulates phosphatidylcholine biosynthesis independent of AT1- and AT2-receptor activation and losartan inhibits phosphatidylcholine biosynthesis by reducing choline uptake in H9c2 cells.
Hatch
G M
GM
Lipid Research Group, Faculty of Medicine, University of Manitoba, Winnipeg, Canada. ghatch@bldghsc.Lan1.umanitoba.ca
Lee
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D
Man
R Y
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Kroeger
E A
EA
Choy
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PC
D000804:Angiotensin II / Q000494:pharmacology; D057911:Angiotensin Receptor Antagonists; D001713:Biphenyl Compounds / Q000494:pharmacology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002794:Choline / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology*; D015151:Immunoblotting; D007700:Kinetics; D019808:Losartan; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D011725:Pyridines / Q000494:pharmacology; D013777:Tetrazoles / Q000494:pharmacology*; D014316:Tritium
D057911:Angiotensin Receptor Antagonists; D001713:Biphenyl Compounds; D007093:Imidazoles; D010713:Phosphatidylcholines; D011725:Pyridines; D013777:Tetrazoles; D014316:Tritium; D000804:Angiotensin II; C073402:PD 123319; D019808:Losartan; D002794:Choline