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Docosahexaenoic acid reduces adenosine triphosphate-induced calcium influx via inhibition of store-operated calcium channels and enhances baseline endothelial nitric oxide synthase phosphorylation in human endothelial cells

Korean Journal of Physiology & Pharmacology 2019년 23권 5호 p.345 ~ 356
 ( Vu Thom Thi ) - Technical University Dresden Medical Faculty Carl Gustav Carus Department of Physiology

 ( Dieterich Peter ) - Technical University Dresden Medical Faculty Carl Gustav Carus Department of Physiology
 ( Vu Thu Thi ) - Vietnam VNU University of Science Faculty of Biology
 ( Deussen Andreas ) - Technical University Dresden Medical Faculty Carl Gustav Carus Department of Physiology

Abstract


Docosahexaenoic acid (DHA), an omega-3-fatty acid, modulates multiple cellular functions. In this study, we addressed the effects of DHA on human umbilical vein endothelial cell calcium transient and endothelial nitric oxide synthase (eNOS) phosphorylation under control and adenosine triphosphate (ATP, 100 μM) stimulated conditions. Cells were treated for 48 h with DHA concentrations from 3 to 50 μM. Calcium transient was measured using the fluorescent dye Fura-2-AM and eNOS phosphorylation was addressed by western blot. DHA dose-dependently reduced the ATP stimulated Ca2+-transient. This effect was preserved in the presence of BAPTA (10 and 20 μM) which chelated the intracellular calcium, but eliminated after withdrawal of extracellular calcium, application of 2-aminoethoxy-diphenylborane (75 μM) to inhibit store-operated calcium channel or thapsigargin (2 μM) to delete calcium store. In addition, DHA (12 μM) increased ser1177/thr495 phosphorylation of eNOS under baseline conditions but had no significant effect on this ratio under conditions of ATP stimulation. In conclusion, DHA dose-dependently inhibited the ATP-induced calcium transient, probably via store-operated calcium channels. Furthermore, DHA changed eNOS phosphorylation suggesting activation of the enzyme. Hence, DHA may shift the regulation of eNOS away from a Ca2+ activated mode to a preferentially controlled phosphorylation mode.

키워드

Adenosine triphosphate; Calcium; Docosahexaenoic acid; Human umbilical vein endothelial cell; Store-operated calcium channels
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