Nifedipine and Bay K 8644 Induce an Increase of [Ca2+] i and Nitric Oxide in Endothelial Cells

Berkels, Reinhard; Mueller, Andreas; Roesen, Renate; Klaus, W.

doi:10.1177/107424849900400307

Cited by 21 publications

(27 citation statements)

References 38 publications

(39 reference statements)

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“…However, others have reported that nifedipine can increase [Ca 2ϩ ] i by activation of Ca 2ϩ release from intracellular store in smooth muscle cells and extracellular Ca 2ϩ influx in coronary endothelial cells (3,25). The intracellular effects of nifedipine reported may be due to high concentration (10 M) or to the cell type used in the study.…”

Section: Discussionmentioning

confidence: 45%

L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

Yakubu

Leffler

2002

American Journal of Physiology-Cell Physiology

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We investigated the role of intracellular calcium concentration ([Ca2+]i) in endothelin-1 (ET-1) production, the effects of potential vasospastic agents on [Ca2+]i, and the presence of L-type voltage-dependent Ca2+ channels in cerebral microvascular endothelial cells. Primary cultures of endothelial cells isolated from piglet cerebral microvessels were used. Confluent cells were exposed to either the thromboxane receptor agonist U-46619 (1 μM), 5-hydroxytryptamine (5-HT; 0.1 mM), or lysophosphatidic acid (LPA; 1 μM) alone or after pretreatment with the Ca2+-chelating agent EDTA (100 mM), the L-type Ca2+ channel blocker verapamil (10 μM), or the antagonist of receptor-operated Ca2+ channel SKF-96365 HCl (10 μM) for 15 min. ET-1 production increased from 1.2 (control) to 8.2 (U-46619), 4.9 (5-HT), or 3.9 (LPA) fmol/μg protein, respectively. Such elevated ET-1 biosynthesis was attenuated by verapamil, EDTA, or SKF-96365 HCl. To investigate the presence of L-type voltage-dependent Ca2+channels in endothelial cells, the [Ca2+]isignal was determined fluorometrically by using fura 2-AM. Superfusion of confluent endothelial cells with U-46619, 5-HT, or LPA significantly increased [Ca2+]i. Pretreatment of endothelial cells with high K+ (60 mM) or nifedipine (4 μM) diminished increases in [Ca2+]i induced by the vasoactive agents. These results indicate that 1) elevated [Ca2+]i signals are involved in ET-1 biosynthesis induced by specific spasmogenic agents, 2) the increases in [Ca2+]i induced by the vasoactive agents tested involve receptor as well as L-type voltage-dependent Ca2+ channels, and 3) primary cultures of cerebral microvascular endothelial cells express L-type voltage-dependent Ca2+ channels.

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Section: Discussionmentioning

confidence: 45%

L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

Yakubu

Leffler

2002

American Journal of Physiology-Cell Physiology

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“…14 -19 Using an NO-sensitive electrode, we previously demonstrated that nifedipine in the micromolar and submicromolar concentration ranges stimulates the NO release from the native endothelium. 33,34 These findings were obtained after acute exposure to DHP calcium antagonists. Therefore, we investigated whether (1) a longterm treatment (48 hours, entire passage 2) of porcine endothelial cell cultures with nifedipine resulted in a similarly altered NO release and (2) an increased formation and/or an impaired inactivation of NO might be involved by measuring the ecNOS (NOS isoform III) mRNA and protein expression and the antioxidative potency of nifedipine, respectively.…”

Section: Discussionmentioning

confidence: 92%

Nifedipine Increases Endothelial Nitric Oxide Bioavailability by Antioxidative Mechanisms

et al. 2001

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Abstract-Short-term treatment of the endothelium with dihydropyridine calcium antagonists resulted in an increased release in NO that is not due to a modulation of L-type calcium channels, because macrovascular endothelial cells do not express this channel. We investigated whether long-term (48 hours) treatment of porcine endothelial cell cultures with the dihydropyridine calcium antagonist nifedipine resulted in a similar enhanced NO liberation. Regarding to the underlying mechanism, we examined whether (1) nifedipine changed the mRNA and protein levels of the constitutive endothelial NO synthase (NOS) in endothelial cell cultures or (2) nifedipine exerts an NO protective effect via its antioxidative properties, as revealed in a cell culture model and with native endothelium from porcine coronary arteries. ihydropyridine (DHP)-type calcium antagonists are important drugs in the treatment of hypertension and coronary heart disease. They induce their specific pharmacological effects by binding to L-type calcium channels, 1,2 which results in a reduced calcium influx with impaired electromechanical coupling both in vascular smooth muscle cells and in the heart. 3 A few years ago, however, it was observed that removal of the endothelium or blockade of the guanylate cyclase of the vessel wall reduced the efficacy of the DHP-induced vasorelaxation, 4 which indicated an endothelium-responsive cGMP-mediated process as part of the DHP action. Because macrovascular endothelial cells lack voltage-operated L-type calcium channels, 5,6 the DHPs must exert these effects via other mechanisms. In this context, it is worth mentioning that DHPs may also exert antihrombotic 7,8 and antiatherosclerotic 9,10 effects in different experimental and clinical settings, of which the underlying signal transduction remains obscure.With this background, it is tempting to speculate that NO, one of the most prominent endothelium-derived factors, 11,12 which relaxes smooth muscle cells via the cGMP signal cascade, 13 might be involved in these DHP actions. In fact, in various models, evidence has accumulated that DHPs stimulate the endothelial NO release, 14 -19 which may mediate or at least contribute to the abovementioned calcium channel-independent effects.Up to now, however, all of these findings were obtained only after acute exposure to DHPs (lasting minutes to hours). Because patients usually take calcium antagonists for a longer period of time, we investigated whether long-term treatment of endothelial cell cultures with nifedipine may alter the basal endothelial NO release as well as the expression of the constitutive endothelial NO synthase (ecNOS) mRNA and protein. Furthermore, because NO is rapidly deactivated by reactive oxygen species (ROS) 20 and the DHPs may act as scavengers as known from different in vitro models, [21][22][23] we also determined the antioxidative potency of nifedipine in endothelial cell cultures as well as in native cells to reveal a potential NO-protection effect as an underlying mechanism of the increased ...

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“…Porcine aortic endothelial cells (PAECs) were enzymatically (dispase, 1 mg/ml; Boehringer Mannheim, Mannheim, Germany) isolated from segments of porcine thoracic aorta and cultured in M199 (Gibco-Invitrogen, Karlsruhe, Germany) supplemented with 10% (v/v) FCS (Gibco-Invitrogen), 100 mg/l penicillin G, 100 mg/l streptomycin, 2 mmol/l L-glutamine and 15 mmol/l HEPES, as described previously [25]. Experiments were performed with confluent cells in passage 2 seeded on glass coverslips.…”

Section: Measurement Of Intracellular Ca 2+ Levelsmentioning

confidence: 99%

“…This method has been described previously [25,26]. Briefly, PAEC monolayers on coverslips were loaded in M199 with the cell-permeable fluorescent dye fura-2-acetomethoxyester (1 µmol/l) for 30 min at 37°C; de-esterification of fura-2-acetomethoxyester to non-permeable fura-2 was completed in fresh M199 by a further 15 min incubation at 37°C.…”

Section: Measurement Of Intracellular Ca 2+ Levelsmentioning

confidence: 99%

Acute effects of glucose and insulin on vascular endothelium

et al. 2004

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Aims/hypothesis. Chronic exposure to high concentrations of glucose has consistently been demonstrated to impair endothelium-dependent, nitric oxide (NO)-mediated vasodilation. In contrast, several clinical investigations have reported that acute exposure to high glucose, alone or in combination with insulin, triggers vasodilation. The aim of this study was to examine whether elevated glucose itself stimulates endothelial NO formation or enhances insulin-mediated endothelial NO release. Methods. We measured NO release and vessel tone ex vivo in porcine coronary conduit arteries (PCAs). Intracellular Ca 2+ was monitored in porcine aortic endothelial cells (PAECs) by fura-2 fluorescence. Expression of the Na + /glucose cotransporter-1 (SGLT-1) was assayed in PAECs and PCA endothelium by RT-PCR. Results. Stimulation of PCAs with D-glucose, but not the osmotic control L-glucose, induced a transient increase in NO release (EC 50 ≈10 mmol/l), mediated by a rise in intracellular Ca 2+ levels due to an influx from the extracellular space. This effect was abolished by inhibitors of the plasmalemmal Na + /Ca 2+ exchanger (dichlorobenzamil) and the SGLT-1 (phlorizin), which was found to be expressed in aortic and coronary endothelium. Alone, D-glucose did not relax PCA, but did augment the effect of insulin on NO release and vasodilation. Conclusions/interpretation. An increased supply of extracellular D-glucose appears to enhance the activity of the endothelial isoform of nitric oxide synthase by increasing intracellular Na + concentrations via SGLT-1, which in turn stimulates an extracellular Ca 2+ influx through the Na + /Ca 2+ exchanger. This mechanism may be responsible for glucose-enhanced, insulin-dependent increases in tissue perfusion (including coronary blood-flow), thus accelerating glucose extraction from the blood circulation to limit the adverse vascular effects of prolonged hyperglycaemia.

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Nifedipine and Bay K 8644 Induce an Increase of [Ca2+] i and Nitric Oxide in Endothelial Cells

Cited by 21 publications

References 38 publications

L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

Nifedipine Increases Endothelial Nitric Oxide Bioavailability by Antioxidative Mechanisms

Acute effects of glucose and insulin on vascular endothelium

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Nifedipine and Bay K 8644 Induce an Increase of [Ca2+] i and Nitric Oxide in Endothelial Cells

Cited by 21 publications

References 38 publications

L-type voltage-dependent Ca2+ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

L-type voltage-dependent Ca2+ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

Nifedipine Increases Endothelial Nitric Oxide Bioavailability by Antioxidative Mechanisms

Acute effects of glucose and insulin on vascular endothelium

Contact Info

Product

Resources

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L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis

L-type voltage-dependent Ca²⁺ channels in cerebral microvascular endothelial cells and ET-1 biosynthesis