Synthesis of prostaglandins by cultured rat heart myocytes and cardiac mesenchymal cells

doi:10.1016/0022-2828(80)90099-1

Journal of Molecular and Cellular Cardiology

Volume 12, Issue 7, July 1980, Pages 685-700

https://doi.org/10.1016/0022-2828(80)90099-1 Get rights and content

Abstract

Ischemia, trauma and hormonal stimulation elicit the release of prostaglandins (PGs) from the heart. Although PGI₂ is synthesized by coronary arteries, the capacities for PG synthesis of individual types of cells comprising the heart have not been elucidated. Accordingly, synthesis of prostaglandins by cultured rat cardiac myocytes and mesenchymal cells was evaluated by radiochromatography of products obtained by incubating cells with [1-¹⁴C]arachidonate, and verified by assessing the effects of cell incubation medium on platelet aggregation. Cultured mesenchymal cells synthesized PGs E₂, F_2α and 6-keto-F_1α, a metabolite of PGI₂ (2076 ± 183, 1284 ± 158 and 1194 ± 152 dpm/mg protein/30 min, respectively). Medium from mesenchymal cells inhibited platelet aggregation, an effect abolished by preincubating the cells with indomethacin, further indicating that these cells synthesized PGI₂. Cardiac myocytes synthesized PGE₂ and PGF_2α (952 ± 227 and 287 ± 104 dpm/mg protein/30 mins, respectively), but no PGI₂. Medium from myocytes did not inhibit platelet aggregation. Prostaglandins D₂, A₂ and thromboxane were not synthesized by either type of cell. Thus, PGI₂ is synthesized by cardiac mesenchymal cells and the hitherto uncharacterized sources of PGE₂ and PGF_2α found in coronary sinus effluent may include cardiac myocytes as well as mesenchymal cells.

References (46)

N.L. Baenziger et al.
Characterization of prostacyclin synthesis in cultured human arterial smooth muscle cells, venous endothelial cells and skin fibroblasts
Cell
(1979)
N.L. Baenziger et al.
Cultured human skin fibroblasts and arterial cells produce a labile platelet-inhibitory prostaglandin
Biochemical and Biophysical Research Communications
(1977)
A.J. Block et al.
Modification of basal release of prostaglandins from rabbit isolated hearts
Prostaglandins
(1974)
T. Chajek et al.
Rat heart in culture as a tool to elucidate the cellular origin of lipoprotein lipase
Biochimica et biophysica acta
(1977)
A. Fujimoto et al.
Studies in vitro on single beating rat-heart cells. IV. The shift from fat to carbohydrate metabolism in culture
Biochimica et biophysica acta
(1964)
R.J. Gryglewski et al.
Arterial walls are protected against deposition of platelet thrombi by a substance (Prostaglandin X) which they make from prostaglandin endoperoxides
Prostaglandins
(1976)
C.C. Haudenschild et al.
Fine structure of vascular endothelium in culture
Journal of Ultrastructure Research
(1975)
S.L. Hong et al.
Stimulation of prostaglandin synthesis by bradykinin and thrombin and their mechanisms of action on MC5-5 fibroblasts
Journal of Biological Chemistry
(1976)
P.S. Kulkarni et al.
Paradoxical endogenous synthesis of a coronary dilating substance from arachidonate
Prostaglandins
(1976)
H. Kuramitsu et al.
Studies in vitro on single beating rat-heart cells. III. Enzyme changes and loss of specific function in culture
Biochimica et biophysica acta
(1964)

S.C. Lee et al.

Prostaglandin metabolism. I. Cytoplasmic reduced nicotinamide adenine dinucleotide phosphate-dependent and microsomal reduced nicotinamide adenine dinucleotide-dependent prostaglandin E 9-keto reductase activities in monkey and pigeon tissues

Journal of Biological Chemistry

(1974)

J.V. Levy

Chronotropic and inotropic effects of PGE₂ on isolated rat atria from normal and spontaneously hypertensive rats (SHR)

Prostaglandins

(1973)

O.H. Lowry et al.

Protein measurement with the Folin phenol reagent

Journal of Biological Chemistry

(1951)

G.V. Marinetti et al.

The phosphatide composition of rat tissues

Biochimica et biophysica acta

(1958)

M. Masson-Pevet et al.

Collagenase- and trypsin-dissociated heart cells: A comparative ultrastructural study

Journal of Molecular and Cellular Cardiology

(1976)

A. Raz et al.

Characterization of a novel metabolic pathway of arachidonate in coronary arteries which generates a potent endogenous coronary vasodilator

Journal of Biological Chemistry

(1977)

M.W. Seraydarian et al.

In vitro studies of beating heart cells in culture. XII. The utilization of ATP and phosphocreatine in oligomycin and 2-deoxyglucose inhibited cells

Biochimica et biophysica acta

(1969)

M.J. Silver et al.

Arachidonic acid-induced human platelet aggregation and prostaglandin formation

Prostaglandins

(1973)

A. Van der Laarse et al.

The activity of cardio-specific isoenzymes of creatine phosphokinase and lactate dehydrogenase in monolayer cultures of neonatal rat heart cells

Journal of Molecular and Cellular Cardiology

(1979)

R.W. Alexander et al.

Stimulation of prostaglandin E synthesis in cultured human umbilical vein smooth muscle cells

R.W. Alexander et al.

Regulation of postocclusive hyperemia by endogenously synthesized prostaglandins in the dog heart

Journal of Clinical Investigation

(1975)

H.J. Berger et al.

Regional cardiac prostaglandin release during myocardial ischemia in anesthetized dogs

Circulation Research

(1976)

C.R. Birdwell et al.

Identification, localization, and role of fibronectin in cultured bovine endothelial cells

Cited by (58)

Beneficial effects of ω-3 fatty acid treatment on the recovery of cardiac function after cold storage of hyperlipidemic rats
1999, Metabolism: Clinical and Experimental
Cardiac effects of ω-3 polyunsaturated fatty acids (PUFAs) were studied in female Wister rats fed a standard diet (control [C] diet) or a high-cholesterol (HC) diet. Subgroups of rats from these groups were treated with eicosapentaenoic acid-E (EPA) or docosahexaenoic acid-95E (DHA) for 5 weeks. Although plasma total cholesterol (TC) and triglyceride (TG) levels were higher in each group fed the HC diet versus each group fed the C diet, EPA administration with the HC diet (HC + EPA) significantly (P < .05) reduced these levels. An isolated working-heart preparation was used to determine cardiac function. Cardiac output (CO) was lower in rats fed the HC diet and HC + DHA versus any of the groups fed the C diet (P < .05). In addition, left ventricular (LV) maximum differentiation of pressure-time curve ( $dp dt$ ) was lower in the rats fed the HC diet versus any of the C diet groups (P < .05). After evaluation of cardiac function in each rat, the heart was stored in a histidine-tryptophanketoglutarate solution for 8 hours at 4°C. The heart was then reperfused, and recovery of cardiac function was evaluated. No significant differences were observed for post-preservative cardiac function within the C diet groups. However, within the HC diet groups, HC + EPA significantly (P < .05) improved the recovery of cardiac function. In addition, HC + DHA also significantly (P < .05) improved the recovery of coronary flow (CF) and LV $dp dt$ . No significant differences were observed for plasma TC and TG concentrations in the C diet groups. EPA administration significantly decreased cardiac levels of palmitic, oleic, and linoleic acids in the HC diet groups. No significant differences were observed for cardiac levels of free fatty acids (FFAs) within the C diet groups. Cardiac EPA and DHA levels were significantly (P < .05) elevated in EPA- or DHA-treated rats compared with the other diet-fed rats. Cardiac EPA levels were also elevated in DHA-treated rats compared with untreated rats (P < .05). These results suggest that EPA attenuates coronary and myocardial preservation injuries through an increase in serum lipids and an accumulation of myocardial FFAs resulting from a HC diet.
Prostaglandin F<inf>2α</inf> stimulates hypertrophic growth of cultured neonatal rat ventricular myocytes
1996, Journal of Biological Chemistry
Prostaglandin F_2α (PGF_2α) stimulates protein synthesis of skeletal and smooth muscle cells in culture and is elevated in the heart during compensatory growth. We hypothesized that PGF_2α stimulates hypertrophic growth of neonatal rat cardiac myocytes. Prostaglandin F_2α increased [³H]phenylalanine incorporation by cultured ventricular myocytes in a dose-dependent manner (EC₅₀ = 11 nM), suggesting action through a PGF-specific receptor. Semiquantitative reverse transcriptase polymerase chain reaction revealed that PGF receptor mRNA is expressed in ventricular myocytes > A7R5 vascular smooth muscle cells ≫ cardiac fibroblast-like cells. The protein content of cardiomyocyte cultures was increased by 10 nM PGF_2α and 11β-PGF_2α but was unchanged by 10 nM PGD₂, PGE₂, PGF_1α, carbaprostacyclin, U-46619, or 12- or 15-hydroxyeicosatrienoic acid. Stimulation of myofibrillar gene expression by PGF_2α was demonstrated by Northern and Western blot analysis for myosin light chain-2 (MLC-2) and by transient transfection experiments with MLC-2 luciferase expression plasmids. In addition, myofibrillogenesis was increased by PGF_2α as assessed by immunocytochemical staining with MLC-2 antisera. Prostaglandin F_2α did not affect myocyte proliferation or [³H]thymidine incorporation, thus myocyte growth occurred by hypertrophy. Proliferative and hypertrophic growth of cardiac fibroblast-like cells were unaffected by PGF_2α. We conclude that PGF_2α stimulates hypertrophic growth of neonatal rat ventricular myocytes in culture and speculate that PGF_2α plays a role in myocardial adaptation to chronic hypertrophic stimuli, recovery from injury, and cardiac ontogeny.
Neutrophils and myocardial reperfusion injury
1996, Pharmacology and Therapeutics
Ischaemia induces an acute inflammatory response in myocardial tissue with an early phase of neutrophil accumulation, which is accelerated by reperfusion. In experimental models, interventions that deplete neutrophils or inhibit their function cause a significant reduction in myocardial infarct size. These cells, therefore, may exacerbate tissue injury through the release of free radicals and proteolytic enzymes. Neutrophil recruitment depends on the presence of inflammatory mediators. Leukotriene B₄, interleukin 8 and the complement fragment C5a have been implicated in this process. Studies using antibodies to the selectin, integrin and immunoglobulin superfamily adhesion molecules indicate that they also have a crucial role in myocardial neutrophil recruitment.
Oxygen deprivation and reoxygenation augment prostacyclin synthesis in cultured ventricular myocytes
1996, Prostaglandins Leukotrienes and Essential Fatty Acids
Prostacyclin production in cultured cardiomyocytes is not induced by cellular ATP depletion per se, suggesting that the mechanism of ischemic injury is more complex. In the present study we subjected cultured ventricular myocytes to ‘simulated ischemia’ followed by reoxygenation. A slight increase in 6-keto-PGF_1α (the stable metabolite of PGI₂) was found during ‘ischemia’, which continued to increase markedly during reoxygenation. PGE₂ levels were pronouncedly enhanced during ischemia but decreased during reoxygenation, and TXB₂ levels remained undetectable throughout. These findings reflect a cardiomyocyte response to anoxic injury, suggesting that they act to protect against cardiac injury by producing the potent vasodilators PGI₂ and PGE₂ during ischemia and reoxygenation.
High oligomycin concentrations augment 6-keto-PGF<inf>1α</inf> production in ventricular cardiomyocytes
1994, Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
Incubation of cultured ventricular cardiomyocytes with high oligomycin concentrations (100 μg/ml), either alone or combined with 2-deoxyglucose (20 mM), led to the rapid depletion of cellular ATP. Inositol (poly)phosphate production decreased, and 6-keto PGF_1α production was increased. In cells depleted of ATP, either by low oligomycin concentrations or by sodium azide, 6-keto PGF_1α was not appreciably increased. There was a 25% rise in the release of fatty acids from the sn-2 position in glycerophospholipids. We suggest that oligomycin at high concentrations causes the release of free arachidonic acid from phospholipids either by non-PIP₂-specific PLC and DG lipase or by phospholipase D, phosphatidic acid phosphatase and DG lipase. The effect is unrelated to decreased cellular ATP content.
Processing of atriopeptin prohormone by nonmyocytic atrial cells
1992, Biochemical and Biophysical Research Communications
Atriopeptin (AP) is synthesized and stored in the mammalian atria as a 126 amino acid prohormone (AP126). Upon secretion, the prohormone undergoes site specific proteolysis within the atria to yield the carboxyl terminal 28 amino acid hormone (AP28). The atrial cell responsible for AP126 bioactivation has not yet been determined. Primary neonatal rat atrial cell cultures were generated with and without depletion of nonmyocytic cells. The molecular form of AP detected in the conditioned media of mixed cultures was determined to be AP126. Addition of dexamethasone to these cultures resulted in the appearance of a peptide that co-migrated with AP28. In contrast, no AP126 processing was detected in the conditioned media of myocyte enriched cultures when grown in the presence of dexamethasone. Readdition of nonmyocytic atrial cells to myocyte enriched cultures successfully reconstituted the steroid induced AP126 processing. Incubation of recombinant AP126argarg with nonmyocytic atrial cell cultures resulted in the generation of AP28argarg. We conclude that a nonmyocytic atrial cell is responsible for AP126 processing in vitro.

View all citing articles on Scopus

^☆: Supported in part by NIH Grants 5-T32-HL-07081, HL-20787, and HL-17646, SCOR in Ischemic Heart Disease.

View full text

Synthesis of prostaglandins by cultured rat heart myocytes and cardiac mesenchymal cells☆

Abstract

Cell

Biochemical and Biophysical Research Communications

Prostaglandins

Biochimica et biophysica acta

Biochimica et biophysica acta

Prostaglandins

Journal of Ultrastructure Research

Journal of Biological Chemistry

Prostaglandins

Biochimica et biophysica acta

Journal of Biological Chemistry

Prostaglandins

Journal of Biological Chemistry

Biochimica et biophysica acta

Journal of Molecular and Cellular Cardiology

Journal of Biological Chemistry

Biochimica et biophysica acta

Prostaglandins

Journal of Molecular and Cellular Cardiology

Stimulation of prostaglandin E synthesis in cultured human umbilical vein smooth muscle cells

Regulation of postocclusive hyperemia by endogenously synthesized prostaglandins in the dog heart

Journal of Clinical Investigation

Regional cardiac prostaglandin release during myocardial ischemia in anesthetized dogs

Circulation Research

Identification, localization, and role of fibronectin in cultured bovine endothelial cells