Metabolism and uptake of adenosine triphosphate and adenosine by porcine aortic and pulmonary endothelial cells and fibroblasts in culture.

Dieterle, Yvonne; Ody, Christiane; Ehrensberger, Alice; Stalder, Hans; Junod, A.

doi:10.1161/01.res.42.6.869

Cited by 78 publications

(44 citation statements)

References 44 publications

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“…There are, in fact, at least three ways in which endothelium could modify platelet behaviour, which are rele-9 vant here. Firstly, the breakdown and, thus, inactivation of pro-aggregatory ADP released from aggregating platelets; secondly, the formation, by hydrolysis of AMP produced via ADP-ase, of anti-aggregatory adenosine, and thirdly, the removal of adenosine by uptake into endothelium [4,5,11,12]. The first two reactions would have anti-aggregatory effects and the third, by diminishing the anti-aggregatory factors in the vascular space, a pro-aggregatory effect.…”

Section: Discussionmentioning

confidence: 99%

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Effect of streptozotocin-induced diabetes on the metabolism of ADP, AMP and adenosine in the pulmonary circulation of rat isolated lung

Bakhle

Chelliah

1983

Diabetologia

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Summary.The uptake and metabolism of radioactive ADP, AMP and adenosine was investigated in the pulmonary circulation of lungs taken from rats with streptozotocin-induced diabetes. Between 0.26 and 500 ~tmol/1 ADP was extensively hydrolysed to AMP equally,in control and diabetic lungs. At 1 mmol/1 there was less ADP breakdown in diabetic lungs. Hydrolysis of AMP to adenosine was also less in diabetic lungs at 10 ~tmol/1 and 1 mmol/1 substrate concentration, but adenosine metabolism and uptake at these concentrations was not affected by diabetes. The results indicate that formation of the anti-aggregatory adenosine and, to a lesser extent, breakdown of the pro-aggregatory ADP were decreased in diabetic lungs and may contribute to the platelet hyperreactivity associated with diabetes.Key words: ADP-ase, adenosine, rat, lung, streptozotocin, diabetes, endothelium.In diabetes mellitus, platelet aggregation is enhanced [3,6,7] and such hyperactive platelet function may contribute to the aetiology of associated vascular disease. However, platelet function in vivo is influenced by the anti-aggregatory properties of vascular endothelium, one of which is the breakdown of the pro-aggregatory adenosine diphosphate (ADP) released from the platelet by an ADP-ase activity associated with endothelial cells [8]. Another unrelated anti-aggregatory property, synthesis of prostacyclin, has already been shown to be decreased in the pulmonary circulation of isolated lungs from diabetic rats [15].We have therefore undertaken an investigation of the metabolism of ADP, AMP and adenosine in the pulmonary circulation of lungs from rats made diabetic by streptozotocin. Materials and Methods Preparation of Diabetic RatsMale Wistar rats (130-150g body weight) were made diabetic by a single IP injection of streptozotocin (85 mg/kg) prepared in a salinecitrate buffer (0.9% NaC1 w/v and 0.1 mmol/1 citrate, pH 4). Sex and age-matched control rats received an injection of the saline-buffer at the same time. Twenty days after treatment, blood glucose in the diabetic rats was found to be 30.3 + 0.6 mmol/1 (n = 6) compared with 8.6 _+ 0.5 mmol/1 in the control animals (n = 6) and this higher glucose level was maintained for up to 60 days. Animals were used between 25 and 50 days after treatment. Preparation of LungsAnimals were anaesthetised by IP injection of sodium pentobarbitone (60 mg/kg). The lungs were removed and perfused via the pulmonary artery as described previously [1] with oxygenated (95% 02:5% CO2) Krebs solution (mmol/l: NaHCO3,25; NaCl,120; KC1,4.7; CAC12,2.5; KH2PO4,1.2; MGSO4,1.2; glucose, 5.6) at 37~ and a constant flow rate of 8 ml/min. Infusion of Radiolabelled SubstratesAfter an initial 10-min period of perfusion to clear the lung of blood and to allow for equilibration, radiolabelled 14C or 3H-ADP, AMP or adenosine (0.665 lxCi of 14C or 6.65 ~tCi of 3H per infusion) together with varying amounts of unlabelled substrate were infused at a rate of 0.8 ml/min for 10 s through the pulmonary arterial cannula. Lung effluent was coll...

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Section: Discussionmentioning

confidence: 99%

“…Two of the endothelium's enzymic activities, ADP-ase and AMP-ase [5,12] were decreased in diabetic lungs. This decrease could be due to a lack of enzymic activity, i. e. altered cell membrane or to a lack of endothelium, i. e. less cells.…”

Section: Discussionmentioning

confidence: 99%

Effect of streptozotocin-induced diabetes on the metabolism of ADP, AMP and adenosine in the pulmonary circulation of rat isolated lung

Bakhle

Chelliah

1983

Diabetologia

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“…Endothelial cells have been cultured from veins of human umiibilical cords (12)(13)(14)(15)(16), vena cavae of rabbits (17), andI large arterial vessels of pigs (18)(19)(20) and cows (21)(22)(23)(24). Most cultured endothelial cells retain the features of endotheliumii in vivo, including an epithelioid shape an(V Weibel-Palade bodies in the cytoplasm (25)(26)(27).…”

Section: Introductionmentioning

confidence: 99%

“…Factor VIII antigen is associated with cells fromn human veins (12,15) and with cells from aortas of animials (19,21,24). Angiotensin I convertinig enzyme activity is found in cells from umbilical cord veins (15,16) and also in aortic endothelial cells from various species (20,28,29). Cultured endothelial cells can transport serotonin (22) and adenine nucleotides (20), and they can form prostacyclin in response to mechanical or enzymatic stimulation (30)(31)(32)(33).…”

Section: Introductionmentioning

confidence: 99%

Human Pulmonary Endothelial Cells in Culture

Johnson¹

1980

J. Clin. Invest.

205

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A B S T R A C T Encdothelial cells were cutltured from various different human vessels, including aortas, pulmonarv, ovarian, and umbilical arteries, an(l pulmonary, ovarian, an(l umilbilical veins. The culturedl cells were identifie(d as endothelial cells by the presence of Factor VIII antigeni and antiotenisini I convertinig enzyme (kininlase II). They retainedl these markers for at least five passages in culture, and somle cells had them for seven passages or nmore. Endothelial cells from the various vessels were comiiparedl with respect to their ability to metabolize angiotensins I and II and bradykinin. Cells fromi arteries had three to five timles the angiotenisini I converting enzyme activity as cells from veins. The activity of angiotensinase A (aspartyl ami nopeptidlase) had a similar distribution, andl cells from arteries were conisistenitly more active than cells fromn veins. Cultures of endothelial cells from pulmonary and umbilical vessels formed prostacyclin in responise to mechanical stimulationi. MIedia from cell moniolayers that were subjecte(l to a change of medliumii and gentle agitation inhibited aggregation of humilani platelets. This inhibitory activity was generated within 2-5 mnm, and it was not formed by cells that were treated with indomnethacini or tranylcypromiine. Additioni of prostaglanidin (PG)H2 to indomethacin-treated cells restored the ability to form the inhibitor, but cells treated with tranylcypromine were not responsive to PGH2. In experiments where [ 14C ]arachidonic acid was added to the cells before stimulation, the major metabolite identified by thin-layer chromatography was 6-keto PGF1t. Thus, it appears that pulmonary endothelial cells, as well as umbilical cord cells, can form prostacyclin. In experiments comparing the ability of arterial and venous cells to form prostacyclin, arterial cells were more active than venous cells. These studies of cells from various human vessels suggest that the vascular origin ofculturecl endothelial cells determines how they metabolize vasoactive peptides and form prostacyclin.

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“…We have also studied the effects of dipyridamole and aspirin, both of which are currently undergoing clinical trials as antithrombotic drugs (i.e., the Persantineaspirin reinfarction study). Dipyridamole was of particular interest because it inhibits the intracellular uptake of adenosine in several systems, including cultured endothelial cells (7,8) and lung (9,10). Our results provide additional information on the antithrombotic actions of dipyridamole and also suggest that the ability of the vessel wall to degrade ADP will not be compromised by the use of aspirin as an antithrombotic drug.…”

Section: Introductionmentioning

confidence: 99%

Effects of aspirin and dipyridamole on the degradation of adenosine diphosphate by cultured cells derived from bovine pulmonary artery.

Crutchley¹,

Ryan²,

Ryan³

1980

J. Clin. Invest.

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A B S T R A C T To improve understanding of the mechanisms by which ADP is degraded during passage through the pulmonary vascular bed, we examined cultured endothelial and smooth muscle cells of bovine pulmonary artery for their abilities to metabolize [8-"C]ADP. ADP is rapidly converted to AMP and then to adenosine, hypoxanthine, and inosine. Inosine is the major metabolite produced by endothelial cells. Radioactivity (5-10%) is accumulated intracellularly primarily as ATP. Medium containing 50,.M ADP incubated with endothelial cells rapidly loses its ability to aggregate platelets and becomes antiaggregatory under conditions in which prostacyclin is absent. The antiaggregatory activity is probably the result of accumulated adenosine. 10 ,uM dipyridamole inhibits cellular uptake of radioactivity by >90%, and inosine in the medium is largely replaced by adenosine. This is accompanied by increased antiaggregatory activity ofconditioned medium, which can be matched by authentic adenosine at the same concentration. 1 mM aspirin had no effect on the metabolism of ADP by endothelial cells. Our results suggest: (a) Metabolism of ADP during passage through the lung is mainly the result of endothelial ADPase. (b) ADP released from aggregating platelets can be converted to the antiaggregatory substance, adenosine. Dipyridamole may exert some of its antithrombotic actions by preventing the intracellular uptake ofadenosine, thereby increasing its concentration near the site ofthrombus formation. (c) The ability ofthe vessel wall to degrade ADP should not be compromised by the use of aspirin as an antithrombotic drug. (d) Endothelium may retain some of its antithrombogenicity when prostacyclin generation is impaired.

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Metabolism and uptake of adenosine triphosphate and adenosine by porcine aortic and pulmonary endothelial cells and fibroblasts in culture.

Cited by 78 publications

References 44 publications

Effect of streptozotocin-induced diabetes on the metabolism of ADP, AMP and adenosine in the pulmonary circulation of rat isolated lung

Effect of streptozotocin-induced diabetes on the metabolism of ADP, AMP and adenosine in the pulmonary circulation of rat isolated lung

Human Pulmonary Endothelial Cells in Culture

Effects of aspirin and dipyridamole on the degradation of adenosine diphosphate by cultured cells derived from bovine pulmonary artery.

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