RESUMEN
Neuronal Ca-ATPase has the essential function of keeping intracellular Ca levels in the micromolar range. This is a prerequisite for normal neurotransmission. This study was designed to determine whether Ca-ATPase is a target for docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) action: results show that both these fatty acids are inhibitors of Ca-ATPase activity in synaptosomal membranes isolated from rat cerebral cortex (-65+/-5% at [DHA]=20 microg/ml, -59+/-7% at [EPA]=20 microg/ml). The inhibition caused by EPA, but not that of DHA, could be reversed completely by the addition of calphostin, a protein kinase C blocker. In contrast, DHA could stimulate Ca-ATPase activity (+132+/-5% at [DHA]=30 microg/ml) only in calmodulin-depleted membranes. In addition, Na,K-ATPase (which drives the Na-Ca exchanger) was inhibited by both DHA and EPA, both at 30 microg/ml (-15+/-0.7% and -42+/-1%, respectively). These results suggest a mechanism that explains the dampening effect of omega-3 fatty acids on neuronal activity.
Asunto(s)
ATPasas Transportadoras de Calcio/metabolismo , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/enzimología , Ácidos Grasos Omega-3/farmacología , Animales , Calcio/metabolismo , Calcio/farmacología , Ácidos Docosahexaenoicos/farmacología , Relación Dosis-Respuesta a Droga , Masculino , Naftalenos/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
Duodenal ion transport processes are supported by ATPase enzymes in basolateral membranes of the enterocyte. In vivo studies have shown that long term n-6 poly-unsaturated fatty acid (PUFA) supplementation in rats causes increases in intestinal Ca absorption, coupled with a higher total calcium balance and bone calcium content. The present in vitro study was undertaken to test the effect of arachidonic acid (AA), a highly unsaturated (and thus physiologically potent) member of the n-6 PUFA family, on ATPases in enterocyte basolateral membranes isolated with a sorbitol density gradient procedure. This paper presents results which show that AA inhibits Na+,K+-ATPase in a dose-dependent manner (-67% of basal activity at a concentration of 30 microg/ml, P < 0.005) but that this effect is not mediated by protein kinase C, as shown by the use of the protein kinase C blocker calphostin (0.5 microM). Indomethacin (IDM) at 0.1 mM, a cyclo-oxygenase blocker, could also not reverse the inhibitory effect of AA on Na+,K+-ATPase. Ca2+-ATPase, on the other hand, is not affected significantly (-10%, P > 0.05) by arachidonic acid at 30 microg/ml.