- Email: [email protected]
Tu-P10:512 Nitric oxide in pregnancy
Tues&ty, June 20, 2006: Poster Session PIO Biomarkers for cardiovascular disectse
297
extracellular signal-regulated kinase (ERK) phosphorylation were performed to elucidate the redox-sensitive pathway in ET-1 gene expression. Results: ET-1 gene was induced with leptin as revealed by Northern blotting and promoter activity assay. Leptin increased intracellulax reactive oxygen species (ROS) levels which were inhibited by antioxidants. Antioxidants suppressed leptin-induced ET-1 gene expression and ERK phosphorylation. Co-transfection of dominant negative mutant of Ras, Raf and MEK1 attenuated the leptin-increased ET-1 promoter activity, suggesting that the Ras-Raf-ERK pathway is required for leptin-induced ET-1 gene. Truncation and mutational analysis of the ET-1 gene promoter showed that activator protein-1 (AP-1) binding site was an important cis-element in leptin-induced ET-1 gene expression. Conclusions: Our data suggest that the redox-sensitive ERK pathway plays a role in leptin-induced ET-1 gene expression in rat aortic SMCs. Funding: This study was supported in part with grants from Shin Kong Wu Ho-Su Memorial Hospital.
significant different only between ACS and SA patients, pooled together and controls (p=.002). The NO plasma content was lower in SA (p<.02 vs Controls). The ability of A D M A to discriminate between the two forms of coronary disease was high (area under Receiving Operating Curve = 74%) Conclusions: The methylarginines were elevated and the plasma NO low in chronic form of coronary disease. NO synthesis and related endothelial function seem to be impaired only in the chronic processes Funding: Financial support was received from the Italian Ministry of Health
I Tu-P 10:509 II A T T E N U A T I O N OF A N G I O T E N S I N I I - M E D I A T E D REACTIVE OXYGEN SPECIES GENERATION VIA CARBON MONOXIDE FROM HEME OXYGENASE-1 IN M A C R O P H A G E S
Objectives: The purpose of the study was to investigate the influence of time course changes in Endotelin-1 levels and in the activity of nitric oxide (NO) system on restenosis occurrence in patients after coronary stent implantation. Methods: The study population consisted of 68 patients (56,44-9,4 years) in which a successful coronary stent implantation was performed and who had repeated angiograms at 6-month follow up. In all patients we assessed the drculating levels of Endotelin-1 and Nitrates/Nitrites (NOx)three times: before PCI (I), 24 hours (II) and 1 month after (IV) procedure. NOx levels were used as the approach to assess the activity of the endogenous NO system. We compared circulating levels ET-1 and NOx in patients with or without restenosis and assessed their relationship with the restenosis occurance Results: There were no differences in ET-1 concentration between both groups throughout the study. We did not observed notable differences in NOx baseline concentration but, in respect to patients without restenosis, in which NOx concentration returned to initial values after one month, in restenotic patients NOx levels diminished continuously and significantly during the study (for restenotic and nonrestenotic patients respectively: measurement (meas.) I: 36.64-15.1 vs 40.54-17.1 um/L -NS; meas. II: 27.44-11.6 vs 28.24-18.6 um/L -NS; meas III: 18,04-13,6 vs 33,04-23,3 um/L -p<0,05). Conclusions: The believe that diminishing activity of nitric oxide system may indicate progressive endothelium dysfunction in atherosclerotic patients and can accelerate restenosis cascade after coronary stent implantation.
Y. Kamiyama, K. Ishikawa, A. Kobayashi, S. Kimura, Y. Maxuyama.
Fukushima Medical Universi~, Fukushima, Japan Objectives: Angiotensin II (ATII), which generates reactive oxygen species (ROS) via the activation of NADPH oxidase, is one of the predominant atherogenic factors. We have observed cytoprotective roles of heme oxygenase-1 (HO-1) and its reaction products, carbon monoxide (CO) and bilirubin. Since HO-1 was abundantly expressed in macrophage in atherosclerotic lesions, we examined the roles of HO-1 pathway in macrophages. Methods: Cultured J774 cells were exposed to ATII and resultant ROS generation was detected. Cells were pretreated with heme axginate (HA) and Sn-protoporphyrin (SnPPIX) to activate and inhibit HO. To determine which product of HO is responsible for the biological action, cells were pretreated with CO releasing compound (CORC) or bilirubin before ATII exposure. Involvement of p38MAPK, p42/44MAPK and JNK pathways were examined with respective inhibitors. Results: ROS generated after ATII exposure was reduced after HA and increased after SnPPIX. CORC exhibited dose-dependent reduction of ROS generation. In addition, CORC reduced p47phox induction by ATII while p67phox expression remained unchanged. In contrast, bilirubin did not show apparent effects on ROS generation and p47phox expression. Importantly, these biological effects were strongly abolished by the treatment with SB203580 and partially with PD98059. Conclusions: CO produced from HO-1 attenuated ATII-dependent ROS generation and p47phox induction in macrophages. These effects of CO seem to be mediated through predominantly p38MAPK. These results suggest that the effects on macrophages may conduct anti-oxidative and anti-inflammatory properties of HO- 1. Funding: Grant-in-aid 16590703 from the Ministry of Education, Science and Culture of Japan.
I Tu P 10:510 iI P L A S M A M E T H Y L A R G I N I N E S IN A C U T E A N D CHRONIC CORONARY SYNDROMES 1
V. Cavalca "~, F. Veglia I , I. Squellerio I , G. Marenzi I , E Ravagnani I , M. De Metrio I , E. Tremoli 1"3 .1 Centro Catdiologico Monzino - IRCCS, Milan, Italy:
"-bzst of Cardiology, Universi~ of Milan, Milan, Italy: 3Dept of Pharmacological Science, Universi~ of Milan, Milan, Italy Objective: We investigated the plasma levels of methylaxginines, endogenous inhibitors of nitric oxide (NO) synthesis, in acute and chronic coronary syndromes. Asymmetric dimethylaxginine, is demonstrated to be associated with coronary artery disease but, up to now, no relevant and concordant data were reported in relation to clinical disease presentation. Methods: We measured plasma levels of L-Arginine, ADMA, SDMA and NO in 103 coronary patients, angiographycally documented, (47 with acute coronary syndrome, ACS, 56 with stable angina, SA) and in 49 healthy controls. Data were compared by analysis of covaxiance adjusting for age and gender. Results: A D M A levels were higher in SA than in ACS (p< .0001) and in controls (p=.0008); also the SDMA levels were higher in SA (p<.005 and p=.02 vs ACS and controls respectively). The plasma level of L-Arginine was lower in ACS (p<.0001 vs SA) and the ratio L-Arginine/ADMA was
I
Tu-P10:511 I D I M I N I S H E D A C T I V I T Y OF N I T R I C O X I D E i S Y S T E M IN P A T I E N T S W I T H R E S T E N O S I S A F T E R CORONARY STENT IMPLANTATION A. Derkacz I , M. Protaslewlcz , R. Poreba-, A. Skoczynska-, E Nowicki I , R. Skalik I , W. Mazurek I . 1 Cardiology Department, Medical Universi~ of
Wroclaw, Wroclaw, Poland: 2bztetTtal Medicine Department, Medical Universi~ of Wroclaw, Wroclaw, PolatM
I Tu-P10:512 I N I T R I C O X I D E IN P R E G N A N C Y I 1
V. Maleska Ivanovska I , B. Dejanova I , B. Gerasimova 2 , S. Petrovska I , V. Antevska 1 , K. Zafirovska -. ~ 1btstttute . of Physiology, Skopje, Makedonija:
"~btstitute of Nephrology, Skopje, Makedonija Objective: The objective of this study is to determine the values of NO and its correlation with blood pressure in pregnancy. Metbods: Blood samples from control group, normotensive nongravidas, CG (N=40), normotensive gravidas, NT (N=47), hypertensive, HT (N=18), gravidas with pregnancy induced hypertension, PIH (N=25), hypertensive gravidas combined with pregnancy induced hypertension, SH (N=21) and nongravidas with essential hypertension, HTN (N=26) were evaluated for NO. In the same gravidas and nongravidas we studied 24-hour blood pressure. To determine the levels of NO we used the spectrophotometric assay kit based on Conrad method (OXIS, Portland, USA). Results: In NT the level of NO (44.78±261~mol/L) is statistically different compared with control group (40.33±121~mol/L). The level of NO in SH decreased to 25.23±161~mol/L, and is statistically significant v.s. the level in the CG (p<0.017). The lowest level of NO is found in HTN, 224-111~mol/L that is significantly different compared with: CG (p<0.0001); NT (p<0.0007) and HT (p<0.032). The highest value of systolic blood pressure is found in the HTN (133-t-11mmHg) and in SH (1384-16 mmHg). C o n t u s i o n s : Our data indicate that the level of NO increases in normotensive gravidas, but decreases in all types of hypertension, especially at the end of the pregnancy. NO has physiological effect in normotension maintenance, which suggest that NO may be used as predictor in prevention and evaluation of hypertension during pregnancy. Funding: Institute of Physiology, Medical Faculty, Skopje, R.Macedonia
XIV bztetTtational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006
297
extracellular signal-regulated kinase (ERK) phosphorylation were performed to elucidate the redox-sensitive pathway in ET-1 gene expression. Results: ET-1 gene was induced with leptin as revealed by Northern blotting and promoter activity assay. Leptin increased intracellulax reactive oxygen species (ROS) levels which were inhibited by antioxidants. Antioxidants suppressed leptin-induced ET-1 gene expression and ERK phosphorylation. Co-transfection of dominant negative mutant of Ras, Raf and MEK1 attenuated the leptin-increased ET-1 promoter activity, suggesting that the Ras-Raf-ERK pathway is required for leptin-induced ET-1 gene. Truncation and mutational analysis of the ET-1 gene promoter showed that activator protein-1 (AP-1) binding site was an important cis-element in leptin-induced ET-1 gene expression. Conclusions: Our data suggest that the redox-sensitive ERK pathway plays a role in leptin-induced ET-1 gene expression in rat aortic SMCs. Funding: This study was supported in part with grants from Shin Kong Wu Ho-Su Memorial Hospital.
significant different only between ACS and SA patients, pooled together and controls (p=.002). The NO plasma content was lower in SA (p<.02 vs Controls). The ability of A D M A to discriminate between the two forms of coronary disease was high (area under Receiving Operating Curve = 74%) Conclusions: The methylarginines were elevated and the plasma NO low in chronic form of coronary disease. NO synthesis and related endothelial function seem to be impaired only in the chronic processes Funding: Financial support was received from the Italian Ministry of Health
I Tu-P 10:509 II A T T E N U A T I O N OF A N G I O T E N S I N I I - M E D I A T E D REACTIVE OXYGEN SPECIES GENERATION VIA CARBON MONOXIDE FROM HEME OXYGENASE-1 IN M A C R O P H A G E S
Objectives: The purpose of the study was to investigate the influence of time course changes in Endotelin-1 levels and in the activity of nitric oxide (NO) system on restenosis occurrence in patients after coronary stent implantation. Methods: The study population consisted of 68 patients (56,44-9,4 years) in which a successful coronary stent implantation was performed and who had repeated angiograms at 6-month follow up. In all patients we assessed the drculating levels of Endotelin-1 and Nitrates/Nitrites (NOx)three times: before PCI (I), 24 hours (II) and 1 month after (IV) procedure. NOx levels were used as the approach to assess the activity of the endogenous NO system. We compared circulating levels ET-1 and NOx in patients with or without restenosis and assessed their relationship with the restenosis occurance Results: There were no differences in ET-1 concentration between both groups throughout the study. We did not observed notable differences in NOx baseline concentration but, in respect to patients without restenosis, in which NOx concentration returned to initial values after one month, in restenotic patients NOx levels diminished continuously and significantly during the study (for restenotic and nonrestenotic patients respectively: measurement (meas.) I: 36.64-15.1 vs 40.54-17.1 um/L -NS; meas. II: 27.44-11.6 vs 28.24-18.6 um/L -NS; meas III: 18,04-13,6 vs 33,04-23,3 um/L -p<0,05). Conclusions: The believe that diminishing activity of nitric oxide system may indicate progressive endothelium dysfunction in atherosclerotic patients and can accelerate restenosis cascade after coronary stent implantation.
Y. Kamiyama, K. Ishikawa, A. Kobayashi, S. Kimura, Y. Maxuyama.
Fukushima Medical Universi~, Fukushima, Japan Objectives: Angiotensin II (ATII), which generates reactive oxygen species (ROS) via the activation of NADPH oxidase, is one of the predominant atherogenic factors. We have observed cytoprotective roles of heme oxygenase-1 (HO-1) and its reaction products, carbon monoxide (CO) and bilirubin. Since HO-1 was abundantly expressed in macrophage in atherosclerotic lesions, we examined the roles of HO-1 pathway in macrophages. Methods: Cultured J774 cells were exposed to ATII and resultant ROS generation was detected. Cells were pretreated with heme axginate (HA) and Sn-protoporphyrin (SnPPIX) to activate and inhibit HO. To determine which product of HO is responsible for the biological action, cells were pretreated with CO releasing compound (CORC) or bilirubin before ATII exposure. Involvement of p38MAPK, p42/44MAPK and JNK pathways were examined with respective inhibitors. Results: ROS generated after ATII exposure was reduced after HA and increased after SnPPIX. CORC exhibited dose-dependent reduction of ROS generation. In addition, CORC reduced p47phox induction by ATII while p67phox expression remained unchanged. In contrast, bilirubin did not show apparent effects on ROS generation and p47phox expression. Importantly, these biological effects were strongly abolished by the treatment with SB203580 and partially with PD98059. Conclusions: CO produced from HO-1 attenuated ATII-dependent ROS generation and p47phox induction in macrophages. These effects of CO seem to be mediated through predominantly p38MAPK. These results suggest that the effects on macrophages may conduct anti-oxidative and anti-inflammatory properties of HO- 1. Funding: Grant-in-aid 16590703 from the Ministry of Education, Science and Culture of Japan.
I Tu P 10:510 iI P L A S M A M E T H Y L A R G I N I N E S IN A C U T E A N D CHRONIC CORONARY SYNDROMES 1
V. Cavalca "~, F. Veglia I , I. Squellerio I , G. Marenzi I , E Ravagnani I , M. De Metrio I , E. Tremoli 1"3 .1 Centro Catdiologico Monzino - IRCCS, Milan, Italy:
"-bzst of Cardiology, Universi~ of Milan, Milan, Italy: 3Dept of Pharmacological Science, Universi~ of Milan, Milan, Italy Objective: We investigated the plasma levels of methylaxginines, endogenous inhibitors of nitric oxide (NO) synthesis, in acute and chronic coronary syndromes. Asymmetric dimethylaxginine, is demonstrated to be associated with coronary artery disease but, up to now, no relevant and concordant data were reported in relation to clinical disease presentation. Methods: We measured plasma levels of L-Arginine, ADMA, SDMA and NO in 103 coronary patients, angiographycally documented, (47 with acute coronary syndrome, ACS, 56 with stable angina, SA) and in 49 healthy controls. Data were compared by analysis of covaxiance adjusting for age and gender. Results: A D M A levels were higher in SA than in ACS (p< .0001) and in controls (p=.0008); also the SDMA levels were higher in SA (p<.005 and p=.02 vs ACS and controls respectively). The plasma level of L-Arginine was lower in ACS (p<.0001 vs SA) and the ratio L-Arginine/ADMA was
I
Tu-P10:511 I D I M I N I S H E D A C T I V I T Y OF N I T R I C O X I D E i S Y S T E M IN P A T I E N T S W I T H R E S T E N O S I S A F T E R CORONARY STENT IMPLANTATION A. Derkacz I , M. Protaslewlcz , R. Poreba-, A. Skoczynska-, E Nowicki I , R. Skalik I , W. Mazurek I . 1 Cardiology Department, Medical Universi~ of
Wroclaw, Wroclaw, Poland: 2bztetTtal Medicine Department, Medical Universi~ of Wroclaw, Wroclaw, PolatM
I Tu-P10:512 I N I T R I C O X I D E IN P R E G N A N C Y I 1
V. Maleska Ivanovska I , B. Dejanova I , B. Gerasimova 2 , S. Petrovska I , V. Antevska 1 , K. Zafirovska -. ~ 1btstttute . of Physiology, Skopje, Makedonija:
"~btstitute of Nephrology, Skopje, Makedonija Objective: The objective of this study is to determine the values of NO and its correlation with blood pressure in pregnancy. Metbods: Blood samples from control group, normotensive nongravidas, CG (N=40), normotensive gravidas, NT (N=47), hypertensive, HT (N=18), gravidas with pregnancy induced hypertension, PIH (N=25), hypertensive gravidas combined with pregnancy induced hypertension, SH (N=21) and nongravidas with essential hypertension, HTN (N=26) were evaluated for NO. In the same gravidas and nongravidas we studied 24-hour blood pressure. To determine the levels of NO we used the spectrophotometric assay kit based on Conrad method (OXIS, Portland, USA). Results: In NT the level of NO (44.78±261~mol/L) is statistically different compared with control group (40.33±121~mol/L). The level of NO in SH decreased to 25.23±161~mol/L, and is statistically significant v.s. the level in the CG (p<0.017). The lowest level of NO is found in HTN, 224-111~mol/L that is significantly different compared with: CG (p<0.0001); NT (p<0.0007) and HT (p<0.032). The highest value of systolic blood pressure is found in the HTN (133-t-11mmHg) and in SH (1384-16 mmHg). C o n t u s i o n s : Our data indicate that the level of NO increases in normotensive gravidas, but decreases in all types of hypertension, especially at the end of the pregnancy. NO has physiological effect in normotension maintenance, which suggest that NO may be used as predictor in prevention and evaluation of hypertension during pregnancy. Funding: Institute of Physiology, Medical Faculty, Skopje, R.Macedonia
XIV bztetTtational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006