Human vascular smooth muscle cells inhibit platelet aggregation when incubated with glyceryl trinitrate: evidence for generation of nitric oxide

Benjamin, Nigel; Dutton, Jane A. E.; Ritter, James M.

doi:10.1111/j.1476-5381.1991.tb12264.x

Cited by 52 publications

(20 citation statements)

References 15 publications

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“…Besides the well established metabolism of GTN to NO by SMC, EC can also make this conversion (Benjamin et al, 1991;Feelisch & Kelm, 1991;Salvemini et al, 1992 from GTN by SMC or EC is mediated by an enzymatic mechanism has now been substantiated. Thus we (Salvemini et al, 1992) and others (Feelisch & Kelm, 1991) have demonstrated that boiled SMC or EC from bovine or porcine aortae lose about 80-90% of their capacity to form NO from GTN.…”

Section: Discussionmentioning

confidence: 99%

“…Apart from the metabolism of GTN to NO by SMC (Benjamin et al, 1991;Feelish & Kelm, 1991;Salvemini et al, 1992) we and others (Feelish & Kelm, 1991;Salvemini et al, 1992) (GST) and glutathione reductase . Recent 'PI Macmillan Press Ltd, 1993 evidence in hepatic and renal cells has suggested that the cytochrome P4m enzyme system is involved in the formation of NO from GTN (Servent et al, 1989;Schroder & Schrdr, 1992).…”

Section: Introductionmentioning

confidence: 99%

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Conversion of glyceryl trinitrate to nitric oxide in tolerant and non‐tolerant smooth muscle and endothelial cells

Salvemini

Pistelli

Vane

1993

British J Pharmacology

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Exposure of smooth muscle cells (SMC) to glyceryl trinitrate (GTN, 75–600 μm) for 30 min led to a concentration‐dependent increase in nitrite (NO2−), one of the breakdown products of nitric oxide (NO). This was not affected by 30 min pretreatment of the cells with 0.5 mm of sulphobromophthalein (SBP) an inhibitor of glutathione‐S‐transferase (GST), by metyrapone or SKF‐525A inhibitors of cytochrome P450. These experiments were confirmed by organ bath studies using rabbit aortic strips denuded of endothelium and contracted with phenylephrine. Thus, a 30 min incubation of the strips with 0.5 mm SPB, metyrapone or SKF‐525A did not affect the relaxations in response to GTN (10−10−10−6 m). Potentiation of the anti‐platelet effect of GTN (44 μm) by endothelial cells (EC, 40 × 103 cells) was not affected by prior incubation of EC with SBP, metyrapone or SKF‐525A (all at 0.5 mm). Potentiation of the antiplatelet activity of GTN (11–352 μm) by small numbers of SMC (24 × 103 cells) or EC (40 × 103 cells) treated with indomethacin (10 μm) was attenuated when the SMC or EC were treated in culture with a high concentration of GTN (600 μm) for 18 h beforehand (referred to as ‘tolerant’ cells). In addition, tolerant SMC produced far less NO2− than non‐tolerant SMC. Exposure of non‐tolerant SMC or EC (105 cells) to GTN (200 μm) for 3 min increased (3–4 fold) the levels of guanosine 3′:5′‐cyclic monophosphate (cyclic GMP). This increase was much less (≤ 1 fold) in the tolerant SMC or EC (105 cells). The basal levels of cyclic GMP were similar in normal or tolerant SMC or EC. Sodium nitroprusside (80 μm) or atrial natriuretic factor (ANF, 10−7 m) increased the levels of cyclic GMP in normal or tolerant SMC or EC to the same extent. The anti‐platelet effects of GTN (44 μm) were potentiated by the sulphydryl donor N‐acetylcysteine (NAC, 0.5 mm). Incubation of GTN (150–1200 μm) for 30 min with NAC (0.1–1 mm) led to a concentration‐dependent increase in NO2− formation. The reduced ability of tolerant SMC or EC to potentiate the anti‐platelet activity of GTN was restored by NAC (0.5 mm). These anti‐aggregatory effects were abolished by concurrent co‐incubation with oxyhaemoglobin (10 μm) indicating that they were due to NO release. Thus, in SMC or EC, metabolism of GTN to NO does not depend on glutathione‐S‐transferase or the cytochrome P450 system. Furthermore, when compared to normal cells, tolerant SMC or EC metabolize GTN to NO less effectively as assessed by the reduced capacity to potentiate the antiplatelet effects of GTN, to release NO2− and to increase the level of cyclic GMP. This decrease in NO formation shows that tolerance to GTN is mainly due to impaired biotransformation of GTN to NO. NAC, by directly forming NO from GTN, compensates for this failing mechanism.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Conversion of glyceryl trinitrate to nitric oxide in tolerant and non‐tolerant smooth muscle and endothelial cells

Salvemini

Pistelli

Vane

1993

British J Pharmacology

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“…One is enzymatic and the other is nonenzymatic and relies on the presence of thiol-containing molecules such as cysteine. Smooth muscle cells, endothelial cells, and inflammatory cells such as macrophages are capable of converting organic nitrates to NO (Benjamin et al, 1991;Feelisch and Kelm, 1991;Salvemini et al, 1992a,b). In mammals, denitration and clearance of the drug are mainly carried out by cytosolic hepatic glutathione S-transferase enzymes and glutathione reductase (Needleman and Johnson, 1973).…”

Section: Nitric Oxide Donorsmentioning

confidence: 99%

“…Platelets do not have the enzyme that converts organic nitrates to NO, and therefore, in vitro they will respond only to very high doses of nitrates (Ahlner et al, 1991). The fact that endothelial cells can metabolize organic nitrates such as GTN efficiently, as do smooth muscle cells (Benjamin et al, 1991;Feelisch and Kelm, 1991;Salvemini et al, 1992a), is important because it provides the key for understanding why organic nitrates inhibit platelet function in vivo, whereas they generally display weak antiplatelet effects in vitro (Schafer et al, 1980;Loscalzo, 1981;Benjamin et al, 1991;Salvemini et al, 1992a). The findings that smooth muscle cells and endothelial cells significantly increase the in vitro antiplatelet effects of GTN (where the dose of GTN required to inhibit platelet aggregation in vitro approximates that used in in vivo studies) led to the proposal that the increased effectiveness of organic nitrates in vivo as antiplatelet agents is due to conversion to NO by cells of the vascular tree, such as smooth muscle cells and endothelial cells (Benjamin et al, 1991;Feelisch and Kelm, 1991;Salvemini et al, 1992a).…”

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confidence: 99%

Modulation of Prostaglandin Biosynthesis by Nitric Oxide and Nitric Oxide Donors

Muscoli²,

et al. 2005

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“…Nitrates have been found to attenuate results in terms of antiplatelet effects. Intravenous infusion platelet aggregation in vitro at 'supratherapeutic' of nitroglycerin failed to inhibit platelet aggregation in concentrations (Gerzer et al, 1988;Mehta & Mehta, vitro induced by various agonists in patients with pre-1980; Schafer et al, 1980), and interestingly, the presence sumed coronary artery disease (Stamler et al, 1988), of endothelial or smooth muscle cells augments this effect patients with heart failure (Mehta & Mehta, 1980) and (Benjamin et al, 1991;Rolland et al, 1987). The latter healthy volunteers (Fitzgerald et al, 1984), whereas finding suggests that effects of nitrates on platelet platelet inhibiting effects were found in patients with…”

Section: Introductionmentioning

confidence: 99%

Effects of an oral dose of isosorbide dinitrate on platelet function and fibrinolysis in healthy volunteers.

Wallén

Larsson

Bröijersén

et al. 1993

Brit J Clinical Pharma

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1. A randomised double‐blind placebo‐controlled study was performed to investigate the effects of isosorbide dinitrate (ISDN; 20 mg orally) on various aspects of platelet function and fibrinolysis in vivo in 12 healthy volunteers. 2. Measurements were performed at rest (before and after tablet ingestion) and during platelet activation by adrenaline (0.4 nmol kg‐1 min‐1; 30 min infusion). 3. At rest, ISDN did not alter plasma concentrations of beta‐thromboglobulin (beta TG). EC50 values for ADP induced aggregation in vitro (Born aggregometry) or ex vivo filtragometry readings. Adrenaline markedly increased platelet aggregability in vivo as measured by filtragometry and elevated levels of beta TG in plasma. ISDN treatment did not affect these responses in the group as a whole. 4. Individuals responding to ISDN with more pronounced vasodilatation at rest showed a lesser increase in aggregability during the ensuing adrenaline infusion (r = ‐0.66, P = 0.02) despite higher adrenaline levels during ISDN. In individuals showing a significant decrease in systolic blood pressure (n = 8) ISDN tended to attenuate the adrenaline induced increase in platelet aggregability (filtragometry; P = 0.08), despite higher plasma adrenaline and noradrenaline levels after ISDN ingestion. 5. Plasma concentrations of ISDN and its active metabolites isosorbide‐5‐ mononitrate and isosorbide‐2‐mononitrate were not correlated to haemodynamic or platelet variables. 6. Fibrinolytic activity (t‐PA antigen and activity, PAI‐1 antigen and activity) increased similarly during the adrenaline infusion following ISDN and placebo. 7. It is concluded that ISDN may affect platelet aggregation responses to adrenaline in vivo, but only in individuals showing significant haemodynamic responses to ISDN.(ABSTRACT TRUNCATED AT 250 WORDS)

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Human vascular smooth muscle cells inhibit platelet aggregation when incubated with glyceryl trinitrate: evidence for generation of nitric oxide

Cited by 52 publications

References 15 publications

Conversion of glyceryl trinitrate to nitric oxide in tolerant and non‐tolerant smooth muscle and endothelial cells

Conversion of glyceryl trinitrate to nitric oxide in tolerant and non‐tolerant smooth muscle and endothelial cells

Modulation of Prostaglandin Biosynthesis by Nitric Oxide and Nitric Oxide Donors

Effects of an oral dose of isosorbide dinitrate on platelet function and fibrinolysis in healthy volunteers.

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