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Adenine nucleotide metabolism in human endothelial cells
J Mol
Cell
Cardiol
24 (Supplement
V)(1992)
October 2,1992 Neutrophils and Growth Factor Activation 78
Oral presentation During Myocardial &hernia
III: 14.00 - 15.40
EXPRESSION OF MITOGENIC FACTORS IN ISCHEMIC PIG HEART Rem2 Zimmermann’, Angelika Kluge, Masahiro Mohri, Stefan Sack, Pieter D. Verdouw”, Hari S. Sharma and Wolfgang Schaper, Max-Planck-Institute, and ‘Kerckhoff-Klinik, Bad Nauheim; FRGermany, **Erasmus University, Rotterdam, The Netherlands Angiogenesis in the pig heart may be induced by brief coronary occlusions. We therefore studied the expression of the mitogenic factors interleukin-la (IL-la), interleukin-1 O(ll-1 6). interleukin-6 (IL-6), insulin-like growth factor-l (IGF-1) and insulin-like growth factor-2 (IGF-2) in ischemic and stunned porcine myocardium compared to normal myocardium using Northern blot analysis with cDNA probes of these genes. The results showed a complicated pattern of gene expression in porcine heart: IL-la and IL-lb were neither expressed in stunned or ischemic porcine myocardium derived from microembolized or ameroid constricted pigs, nor in normal control myocardium. IL-6 mRNA was expressed at low levels in most of the myocardial tissue studied so far, showing no difference between control and experimental regions. IGF-1 shows a weak but constantly expressed signal for a mRNA of =6.3kb which is not markedly changed between control and experimental areas, whereas at least 4 different mRNAs were observed for IGF-2 in pig heart, the strongest signal appearing at =2.8kb. The expression of IGF-2 mRNAs was high in pigs subjected to a stunning protocol
but weaker in microembolized or ameroid pigs. These findings indicate that the expression 11-6, IGF-1 and IGF-2 in the ischemic myocardium may contribute to angiogenic processes. 135
ADENINE Ryszard Magdi
NUCLEOTIDE
METABOLISM
IN HUMAN
T: Smolenski, Zdzislaw Kochan, Chris H. Yacoub, M. R. S. Unit, National Heart end
Page, Rhode Lung Institute
ENDOTHELIAL McDoual, et Harefield
of
CELLS Anne-Marie Hospital,
1. Seymour, Herefield, U.K.
Knowledge of endothelial adenylate metabolism is important for evaluation of the role of endothelium in myocardial nucieotide metabolism and physiological actions. We have investigated the metabolism of cultured endothelial cells from human umbilical cord under control conditions (normal oxygen and substrate supply) and under conditions of ATP starvation, in the presence of inhibitors of glycolysis and mitochondrial ATP formation, iodoacetate and oligomycin (I+ 0). Inhibitors of adenosine deaminase (EHNA) and adenosine kinase 5’-iodotubercidin (ITu) were included in some experiments to define pathways of nucleotide catabolism. Purine metabolites were evaluated in the cells and in the medium after 45 min of incubation at 37°C and the total amounts per culture flasks were as follows: Under control conditions in the presence of EHNA, ATP in the cells was 9 nmol, whilest in the medium hypoxanthine and xanthine increased by 1.5 nmol and uric acid by 0.3 nmol. If ITu was also present, adenosine formation was observed (0.8 nmol). In the presence of I+0 and EHNA almost complete ATP depletion was observed after 45 min. AMP increased to 3 nmol, adenosine content was 6 nmol and (0.4 nmol). endothelial presence of nucleotides
158
inosine+ hypoxanthine-txanthine We conclude that adenosine cells under normal oxygenation xanthine oxidoreductase but are catabolised predominantly
was 2.5 nmol. Traces of uric acid were also detected cycling (formation and reincorporation) can operate in the and substrate supply. Formation of uric acid suggests the this activity appears to be very small. In energy deprived cells via the pathway involving adenosine formation.
INTRAVENOUS ISOSORBIDE DINITRATE INHIBITS NEUTROPHIL AGGREGATION Tomasz Siminiak, Alina Abramowska, Katarsyna Czechowska, Pawe Prycki, Dorota Zosuli&ka, Henryk Wysocki; Academy of Medicine, Department of Intensive Therapy, Poznan, Poland Due to the role of neutrophils (PMN) in cardiovascular pathology, the recognition of drugs effect on PMN function may be of clinical importance. We have previously reported that 24h oral treatment with isosorbide dinitrate (ISDN) increases PMN tissue migration but does not affect PMN aggregation and adherence. To evaluate the effect of "acute" treatment with ISDN on PMN function we collected blood samples from 17 patients with ischemic heart disease (11 with myocardial infarction and 5 with unstable angina) that underwent routine infusion of intravenous ISDN (Isoket. Schwarz-Pharma). PMN aggregation and adherence were measured with whole blood assays before and at 15 and 30 mip. of drug+infusion. Intravenous ISDN decreased PMN aggregation (25.3-3.6, 19.0-2.6 and 18;5-2.6 respectively, ~$0.01) and did not affect PMN adherence (13.4-2.4, 13.3-3.3 and 15.5-3.3, respectively). s.74
Cell
Cardiol
24 (Supplement
V)(1992)
October 2,1992 Neutrophils and Growth Factor Activation 78
Oral presentation During Myocardial &hernia
III: 14.00 - 15.40
EXPRESSION OF MITOGENIC FACTORS IN ISCHEMIC PIG HEART Rem2 Zimmermann’, Angelika Kluge, Masahiro Mohri, Stefan Sack, Pieter D. Verdouw”, Hari S. Sharma and Wolfgang Schaper, Max-Planck-Institute, and ‘Kerckhoff-Klinik, Bad Nauheim; FRGermany, **Erasmus University, Rotterdam, The Netherlands Angiogenesis in the pig heart may be induced by brief coronary occlusions. We therefore studied the expression of the mitogenic factors interleukin-la (IL-la), interleukin-1 O(ll-1 6). interleukin-6 (IL-6), insulin-like growth factor-l (IGF-1) and insulin-like growth factor-2 (IGF-2) in ischemic and stunned porcine myocardium compared to normal myocardium using Northern blot analysis with cDNA probes of these genes. The results showed a complicated pattern of gene expression in porcine heart: IL-la and IL-lb were neither expressed in stunned or ischemic porcine myocardium derived from microembolized or ameroid constricted pigs, nor in normal control myocardium. IL-6 mRNA was expressed at low levels in most of the myocardial tissue studied so far, showing no difference between control and experimental regions. IGF-1 shows a weak but constantly expressed signal for a mRNA of =6.3kb which is not markedly changed between control and experimental areas, whereas at least 4 different mRNAs were observed for IGF-2 in pig heart, the strongest signal appearing at =2.8kb. The expression of IGF-2 mRNAs was high in pigs subjected to a stunning protocol
but weaker in microembolized or ameroid pigs. These findings indicate that the expression 11-6, IGF-1 and IGF-2 in the ischemic myocardium may contribute to angiogenic processes. 135
ADENINE Ryszard Magdi
NUCLEOTIDE
METABOLISM
IN HUMAN
T: Smolenski, Zdzislaw Kochan, Chris H. Yacoub, M. R. S. Unit, National Heart end
Page, Rhode Lung Institute
ENDOTHELIAL McDoual, et Harefield
of
CELLS Anne-Marie Hospital,
1. Seymour, Herefield, U.K.
Knowledge of endothelial adenylate metabolism is important for evaluation of the role of endothelium in myocardial nucieotide metabolism and physiological actions. We have investigated the metabolism of cultured endothelial cells from human umbilical cord under control conditions (normal oxygen and substrate supply) and under conditions of ATP starvation, in the presence of inhibitors of glycolysis and mitochondrial ATP formation, iodoacetate and oligomycin (I+ 0). Inhibitors of adenosine deaminase (EHNA) and adenosine kinase 5’-iodotubercidin (ITu) were included in some experiments to define pathways of nucleotide catabolism. Purine metabolites were evaluated in the cells and in the medium after 45 min of incubation at 37°C and the total amounts per culture flasks were as follows: Under control conditions in the presence of EHNA, ATP in the cells was 9 nmol, whilest in the medium hypoxanthine and xanthine increased by 1.5 nmol and uric acid by 0.3 nmol. If ITu was also present, adenosine formation was observed (0.8 nmol). In the presence of I+0 and EHNA almost complete ATP depletion was observed after 45 min. AMP increased to 3 nmol, adenosine content was 6 nmol and (0.4 nmol). endothelial presence of nucleotides
158
inosine+ hypoxanthine-txanthine We conclude that adenosine cells under normal oxygenation xanthine oxidoreductase but are catabolised predominantly
was 2.5 nmol. Traces of uric acid were also detected cycling (formation and reincorporation) can operate in the and substrate supply. Formation of uric acid suggests the this activity appears to be very small. In energy deprived cells via the pathway involving adenosine formation.
INTRAVENOUS ISOSORBIDE DINITRATE INHIBITS NEUTROPHIL AGGREGATION Tomasz Siminiak, Alina Abramowska, Katarsyna Czechowska, Pawe Prycki, Dorota Zosuli&ka, Henryk Wysocki; Academy of Medicine, Department of Intensive Therapy, Poznan, Poland Due to the role of neutrophils (PMN) in cardiovascular pathology, the recognition of drugs effect on PMN function may be of clinical importance. We have previously reported that 24h oral treatment with isosorbide dinitrate (ISDN) increases PMN tissue migration but does not affect PMN aggregation and adherence. To evaluate the effect of "acute" treatment with ISDN on PMN function we collected blood samples from 17 patients with ischemic heart disease (11 with myocardial infarction and 5 with unstable angina) that underwent routine infusion of intravenous ISDN (Isoket. Schwarz-Pharma). PMN aggregation and adherence were measured with whole blood assays before and at 15 and 30 mip. of drug+infusion. Intravenous ISDN decreased PMN aggregation (25.3-3.6, 19.0-2.6 and 18;5-2.6 respectively, ~$0.01) and did not affect PMN adherence (13.4-2.4, 13.3-3.3 and 15.5-3.3, respectively). s.74