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Biomedical subjects

M J Adolfs

Publications and source records attributed to M J Adolfs.

At least 37 records · Page 2Linked to original sources

Antiinflammatory aspects of exogenous arachidonic acid.

Resident rat peritoneal macrophages (M0) were more active in metabolising exogenous arachidonic acid (AA) to prostaglandin E2 (PGE2) than starch elicited cells. Basal levels of cAMP were lower in elicited macrophages than in resident cells but the adenyl cyclase of elicited macrophages was much more readily stimulated by cyclooxygenase products formed from exogenous AA than resident cells. AA injected into carrageenin sponge granulomas reduced the severity of the inflammation in a dose related manner.

Animals↗

Regulation of prostaglandin E2 receptors in vivo by dietary fatty acids in peritoneal macrophages from rats.

Groups of rats were pretreated with 4-week diets containing 12.5% corn oil or linseed oil. At the end of this period peritoneal macrophages were elicited and isolated. These cells were used for binding experiments with 3H-PGE2 and for estimation of prostaglandin-stimulated cAMP production. Specific binding of 3H-PGE2 was saturable, reversible, protein-dependent, and correlated with stimulation of cAMP production, indicating that specific binding referred to receptor binding. PGE1 and PGI2 were far less effective than PGE2 in competition of binding with 3H-PGE2, indicating receptor selectivity for PGE2. Scatchard analysis of the specific binding data revealed a high affinity component (Kd 17 nM) and low affinity component. The total number of high- and low-affinity binding sites, respective Kd values, and PG stimulation of cAMP production of cells from rats fed the linseed oil diet were comparable to controls. The corn oil diet, however, resulted in a twofold increase in total number of high- and low-affinity binding sites, while respective Kd values were unchanged. This enhancement of binding capacity could be explained by an increased density of binding sites on the cells, and may itself be responsible for the increased sensitivity of the macrophages in this diet group for PG-stimulated cAMP production. The data suggest a regulatory mechanism at the receptor level and are discussed in terms of possible altered bioavailability of arachidonic acid-derived PGE2.

Animals↗

Direct evidence for the presence of selective binding sites for (3H) prostaglandin E2 on rat peritoneal macrophages.

A method is presented which provides for a simple and rapid determination of PGE2 receptors on viable peritoneal macrophages. Incubation of the harvested cells with (3H)PGE2 revealed specific binding of (3H)PGE2 by use of the Millipore filter assay system. Maximum binding was attained in the presence of 1 mM EDTA. Specific binding was saturable at 65 fmol/mg protein with an equilibrium dissociation constant (Kd) of 3.2 X 10(-8)M. Inhibition of (3H)PGE2 binding with unlabelled prostaglandins revealed a potency series of PGE2 greater than PGE2 greater than PGI2. The PGE2 concentration which displaced 50% of the labelled ligand was 10(-7)M. Comparable kinetic data were obtained for adenylate cyclase stimulation, since the concentration which showed a halfmaximal stimulation of cAMP production was 2 X 10(-7)M of PGE2. Since PGE1 and PGI2 compete with (3H)PGE2 binding in a non-parallel manner compared to PGE2 itself, it is proposed that macrophages possess different types of PG receptors.

Animals↗

Competition for adenyl cyclase coupled (3H)-prostacyclin binding sites with prostaglandin E2 in rat peritoneal macrophages.

Prostaglandins regulate macrophage function by their action on membrane-associated adenyl cyclase. In order to define more directly macrophage-prostaglandin interactions, a binding assay has been developed for macrophage receptors using (3H)-PGI2 as ligand. (3H)-PGI2 binding was specific, saturable and reversible. Moreover, specific binding showed to be enriched in a membrane-enriched fraction of the cells. The assay conditions ensured stability of (3H)-PGI2 during incubations and should exclude intracellular accumulation of the ligand in macrophages. Unlabelled PGE2 and PGI2 competed for (3H)-PGI2 specific binding in both macrophages and membrane preparations. PGE2 showed to be more potent in this respect than PGI2, a phenomena which was also observed for prostaglandin activation of cAMP production in macrophages. The data suggest an interaction at receptor level of endogenously released PGE2 and PGI2 by peritoneal macrophages in vivo and provide support for a previously proposed mechanism of action of low concentrations of PGE2, counteracting stimulation of cAMP production by PGI2 in macrophages.

Adenylyl Cyclases↗

Low concentrations of prostaglandin E2 inhibit the prostacyclin-induced elevation of cyclic adenosine 3',5'-monophosphate in elicited populations of rat peritoneal macrophages.

Elevation of cyclic adenosine 3',5'-monophosphate (cyclic AMP) in elicited populations of rat peritoneal macrophages was used as a parameter to examine the influence of prostaglandin E2 (PGE2) on the effects of prostacyclin (PGI2) and (+/-)-5E-13,14-didehydro-carbo-prostacyclin (DDH-carbo-PGI2) in vitro. PGE2, within the range of 1.4 X 10(-9) to 1.2 X 10(-8)M, caused a concentration-dependent inhibition of the rise in cyclic AMP induced by 2.8 X 10(-6) M PGI2 or DDH-carbo-PGI2. With higher concentrations of PGE2 the inhibition was either non-existent or masked by the effect of PGE2 per se on cyclic AMP levels. The present findings suggest that the earlier observed low responsiveness of granuloma macrophages to PGI2, in terms of rise in cyclic AMP, is possibly due to permanent exposure of these cells to environmental endogenous PGE2.

Animals↗

The stimulation by ethanol of rat aorta ring prostacyclin-like synthesis is related to the age of the animal.

Rat aorta rings were incubated with Tris/HCl buffer and the release of prostacyclin-like material (PGI2-L) measured indirectly by assaying the increase in cAMP in rat platelets incubated with the aorta incubation medium. The addition of 1% ethanol to the incubation system stimulated PGI2-L formation. Aorta rings from 16 week old rats synthesised more PGI2-L material than aorta tissue removed from 6 week old animals.

Age Factors↗

Synergism of (15 S)-15-methyl-prostaglandin E1 with either azathioprine or prednisolone on the survival of heart allografts in rats.

Survival time of Brown Norway (BN) to Wistar (WR) heart allografts without immunosuppressive treatment is 7-10 days. Azathioprine or prednisolone do not improve the graft survival in this model significantly. However, treatment with (15 S)-15-methyl-PGE1 (15-M-PGE1), a synthetic long acting prostaglandin, which can be administered orally, in combination with either azathioprine or prednisolone increased the survival of the allogeneic heart grafts substantially. A synergistic inhibitory effect of 15-M-PGE1 and prednisolone was also noted in carrageenan induced granulomatous inflammation.

Alprostadil↗

Prostaglandin E2 elevation of cyclic-AMP in granuloma macrophages at various stages of inflammation: relevance to anti-inflammatory and immunomodulatory functions.

In the carrageenin-induced granuloma of rats the inflammatory tissue growth and macrophage invasion on the one hand and the cyclic-AMP content of the macrophages on the other, display opposite directional changes. Macrophages, isolated from this tissue at different stages of inflammation, were used to examine the effect of prostaglandin E2 on intracellular levels of c-AMP. It appears that during infiltration of the macrophages into the inflammatory tissue, the sensitivity of adenylate cyclase to activation by PGE2 increases. Arguments are presented that these observations made in vitro, are in direct relevance to the previous described anti-inflammatory effect of PGE on granuloma tissue in vivo.

Animals↗

Distinction between responsiveness of macrophages to cyclic-AMP elevation by prostaglandin E2 and prostacyclin.

Macrophages, isolated from carrageenin-induced granuloma (an immune-related inflammatory model), were used to examine the effects of prostaglandin E2 and prostacyclin on intracellular levels of cyclic-AMP. It appears that during the development of the tissue-growth component of this granuloma in rats the infiltrating mononuclear phagocytes undergo a change, resulting in macrophages which are very sensitive to cyclic-AMP elevation by PGE2 but insensitive to a similar effect of prostacyclin. Arguments are presented that the difference between the responsiveness of these macrophages to PGE2 and prostacyclin observed in vitro is directly related to the previously reported distinction between these two metabolites of arachidonate as inhibitors of the carrageenin-induced granuloma in vivo.

Animals↗

Cyclic AMP in macrophages from experimental granulomas and the effect of prostaglandin E2.

Leucocytes were isolated by pronase digestion from granuloma tissue at different stages in inflammation induced by carrageenin-soaked sponge implants in rats and cyclic AMP was measured in these cells. In a mixed cell suspension, containing granulocytes and macrophages, the cyclic AMP levels increased during the early stages of inflammation but decreased when the granuloma became established. However, after correction for the proportion of infiltrating macrophages, as the inflammation progressed only a fall in cyclic AMP content was observable. Exposure of granuloma-derived cells to PGE2 resulted in a rise in cyclic AMP content, which was more pronounced in cells isolated during a later rather than an earlier stage of granuloma development. The results provide support for the earlier proposal that the anti-inflammatory effect of E-type prostaglandins on granulomas is partially explicable on the basis of cyclic AMP changes in infiltrating macrophages.

Animals↗

Cannulated teflon chamber implant in the rat: a new model for continuous studies on granulomatous inflammation.

A granulomatous inflammation model is described. It is a modification of the cannulated sponge method and consists of a subdermally implanted teflon cylinder in the rat. This 'chamber' is cannulated and provided with holes to ensure contact and exchange of materials between the inner chamber and the surrounding tissues. The cannulae are exterioized at the back of the neck and allow materials to be injected and exudate to be collected at any given time during the development of granulomatous tissue around the chamber. Some parameters of this inflammation, such as exudation, granuloma growth, cellular infiltration in exudate, and the formation of prostaglandins were studied, and a comparison was made with the sponge implant model. The value of the chamber method is illustrated by brief reference to a study on the metabolism of 14C-arachidonic acid during granulomatous inflammation.

Animals↗

Distribution and further studies on the activity of prostaglandin E in chronic granulomatous inflammation.

Pre-formed granulomata, induced by cannulated, carrageenin-soaked, subdermal sponge implants, were used to further analyse the effects of exogenous PGE and the involvement of endogenous PGE in this model of chronic inflammatory tissue changes in rats. Experiments with 14C-PGE2 indicated that administration of PGE into the sponge creates a kinetic situation likely to imitate, but superimposed upon, the continuous discharge of endogenous PGE in the granuloma. The granuloma-reducing, anti-flammatory effect of PGE can be mimicked by dibutyryl cyclic-AMP, phosphodiesterase inhibitors and indomethacin. Most probably these compounds, in common with PGE, exert their anti-granuloma action through elevation of intracellular cyclic-AMP (in macrophages and/or fibroblasts). The anti-granuloma action is not necessarily associated with a reduction in the endogenous PGE-level and vice versa. The involvement of endogenous PGE in the tissue events of granulomatous inflammation is conjectural as yet.

Animals↗