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Tu-P7:254 Matrix metalloproteinases and tissue inhibitor metalloproteinases in hypertensive patients before and after antihypertensive therapy
Tues&ty, June 20, 2006: Poster Session P7 Basic science ( lst part)
240
Conclusion: Hypoxia decreased the biosynthesis of cell associated HSPGs in macrophages. These changes may contribute to motility alterations leading to accumulation and clustering of macrophages in atherosclerotic lesions. Funding: ALF grants of SaJalgrenska University Hospital, the Heart and Lung Foundation, and AstraZeneca.
Tu-P7:251
P L A S M A M A R K E R S OF F I B R O S I S L E V E L S IN H Y P E R T E N S I O N IN C O M B I N A T I O N W I T H ISCHEMIC HEART DISEASE
A. Belovol, O. Kovalyova, E. Kolosov. State Medical Universi~, Kharkov,
Ukrabw Objective: We aimed to estimate the involvement of proform of matrix metallopro-teinase-1 (proMMP-1) and tissue inhibitor of metalloproteinases1 (TIMP-1) in left ventriculax (LV) remodeling in hypertension (HT) in combination with ischemic heart disease (IHD). Methods: 69 patients (pts) with HT in combination with IHD and 20 normotensive (NT) persons were examined. All pts were divided into 4 groups: normal geometry (NG), LV concentric remodeling (CR), LV concentric hypertrophy (CH) and LV ec-centric hypertrophy 0EH). Plasma proMMP-1 (TBS, United Kingdom) and TIMP-1 (BioSource International, Belgium) levels were determined with ELISA. Results: The plasma proMMP-1 level was higher in pts with HT with IHD than in NT persons (3,87-t-2,25 ng/ml versus 1,38-t-0,82 ng/ml, p<0,001). A direct correlation was found between plasma proMMP-1 level and LVMI (r--0,67, p<0,001), LV end dia-stolic-diameter index (r=0,66, p<0,001). Plasma TIMP-1 concentration was higher in pts with HT in combination with IHD than in NT persons (409,23+4,13 ng/ml versus 368,55+7,43 ng/ml, p<0,05). TIMP-1 level correlated with LVMI (r=0,38, p<0,05). Conclusions: Change of proMMP-1 level is compensatory mechanism in response to alteration of LV mass, size and myocardial structure. The excessive proMMP-1 production correlates with progressive increase of LVMI mainly relevant to LV chamber dimensions and less with degree LV wall thickness in patients with HT in combination with IHD. The high TIMP-1 level promotes increase of LVMI at the expense of collagen accumulation in myocardial extracellular matrix as a result of lowering activity of MMP-1.
Tu-P7:252
G R O U P IVA P H O S P H O L I P A S E A 2 - M E D I A T E D P R O D U C T I O N OF F I B R O N E C T I N BY O X I D I Z E D L D L IN M E S A N G I A L C E L L S
S. Akiba, Y. Mukaida, K. Hane, M. Oka, T. Sato. Kyoto Pharmaceutical
Universi~, Kyoto, Japan Objective: The development and progression of glomerulax diseases including glomerulosclerosis and diabetic nephropathy are associated with the accumulation of extracellular matrix proteins 0ECM) and the deposition of atherogenic lipoproteins such as low-density lipoprotein (LDL) and oxidized LDL (oxLDL) within the mesangium. The atherogenic lipoproteins are shown to stimulate the production of fibronectin and collagen, components of ECM, in mesangial cells. To clarify the mechanisms underlying the oxLDL-induced production of ECM, we examined the possible involvement of group IVA phospholipase A2 (PLA2) using human mesangial cells and group IVA PLA2-deficient mouse mesangial cells. Results: oxLDL accelerated the production of fibronectin and collagen (type IV) 6 h and 24 h after the stimulation, respectively. Under the conditions, oxLDL induced the release of axachidonic acid 3 h after the stimulation. Methyl arachidonyl fluorophosphonate (MAFP), an inhibitor of group IVA PLA2, markedly suppressed the oxLDL-induced production of fibronectin as well as the release of arachidonic acid, while it had no effect on the production of collagen. The inhibitory effect of MAFP on the production of fibronectin was reversed by adding arachidonic acid. Furthermore, in group IVA PLA2deficient mouse mesangial cells, the production of fibronectin induced by oxLDL was weak as compared with that in wild-type cells. However, the production by oxLDL of collagen was not affected in the deficient cells. Conclusion: These findings suggest that group IVA PLA2 is involved in the production of fibronectin in oxLDL-stimulated mesangial cells.
I Tu-P7:253 I S U B S T R A T E - D E P E N D E N T
R E G U L A T I O N OF
INFLAMMATORY
S. Filippov, M. Chambers, R. Homan. Esperion Therapeutics, Division of Pfizer Global Research and Development Ann Arbor; Michigan, USA In vulnerable atherosclerotic lesions, the dissolution of extracellulax matrix scaffolds is associated with the local accumulation of macrophages that express heightened levels of a diverse mix of pro-inflammatory cytokines and matrix-degrading enzymes. The regulation of monocyte differentiation by the nature and physical characteristics of the underlying substratum was examined in primary human monocytes that were adhered to either type I collagen or an uncoated plastic surface (representing physiological versus modified matrices, respectively). The protein expression profiles following monocyte to macrophage differentiation were assessed by antibody arrays. In addition to their very distinct shape and morphological appearance, macrophages differentiated on plastic expressed considerably higher levels of pro-inflammatory cytokines. Several of these, including IL-6, IL-8, MCP-1, MIP-lb, TNF-a and eotaxin-2, axe associated with lesion vulnerability in vivo. The macrophages exposed to the rigid surface of uncoated plastic consistently displayed marked upregulation of neutrophil collagenase (MMP-8), along with its most potent physiological activator stromelysin 2 (MMP- 10). Interestingly, the expression of other secreted members of matrix metalloproteinase family, MMP-1, MMP-2, MMP-3 and MMP-13, remained scarcely detectable under both culture conditions. Thus, adhesive interactions with the substratum direct human monocyte-macrophage differentiation and regulate macrophage tissue-destructive potential via expression of pro-inflammatory cytokines and the MMP-8/MMP- 10 axis.
Tu-P7:254
MATRIX METALLOPROTEINASES AND TISSUE I N H I B I T O R M E T A L L O P R O T E I N A S E S IN HYPERTENSIVE PATIENTS BEFORE AND AFTER ANTIHYPERTENSIVE THERAPY
G. Derosa 1 , S. Salvadeo 1 , A. D•Angelo 1 , C. Tinelli 2 , A.F.G. Cicero 3 , L. CiccaxelliI , M.N. Piccinni I , F. Pricolo I , A. Gravina I , R. Fogari I .
1Department of bttental Medicine attd Therapeutics, Universi~ of Pm'ia, Pcn'ia, Italy." 2Bionwtric Unit, IRCCS Policlinico San Matteo, Pavia, Italy." 3Departnwnt of Clinical Medicine attd Applied Biotechnology, Universi~ of Bologna, Bologna, Italy Aim: To test the hypothesis that MMP-2, MMP-9 and TIMP-1 axe altered in hypertension reflecting alterations in E C M turnover and to assess whether chronic antihypertensive treatment with doxazosin would normalize these alterations. M a t e r i a l s a n d M e t h o d s : We enrolled 44 hypertensive patients before and after 4 months treatment with doxazosin. We measured fasting plasma glucose (FPG), fasting plasma insulin (FPI), homeostasis model assessment index (HOMA index), systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), triglycerides (Tg), plasminogen activator inhibitor-1 (PAI-1), homocysteine (Hct) fibrinogen (Fg), C-reactive protein (hs-CRP), and plasma levels and activities of MMP-2, MMP-9 and TIMP- 1. Results: Significant FPI value decrease was observed in treated hypertensive group. SBP and DBP decrease was obtained in treated hypertensive group. Significant hs-CRP value was decreased in treated hypertensive patients. MMP-2 and MMP-9 levels and activity, and TIMP-1 were significantly lower in treated hypertensive group. Conclusions: Plasma levels and activities of MMP-2, MMP-9, and TIMP1 axe decreased in treated hypertensive patients compared to values before treatment.
ITu-P7:2551 MI NAHTIRB II XT OMREMT AELTLAOL PLROOP RT EOITNEAI SNEASS EASN DINT I S S U E HYPERTENSIVE PATIENTS •
l
.
.o
G. Derosa 1, A. D Angelo , C. Tinelh-, L. Ciccaxelli1 , M. Piccinni 1 , F. Pricolo I , S. Salvadeo I , M. Ghelfi I , I. Ferrari I , A. Cicero 3 . 1Department of
bttetTtal Medicine arm Therapeutics, Universi~ of Pavia, Pm,ia, Italy: 2Biometric Unit, IRCCS Policlinico San Matteo, Pavia, Italy." - Department of Clbtical Medicbw arm Applied Biotechnology, Universi~ of Bologna, Bologna, Italy Aim: We hypothesized that MMP-2, MMP-9 and TIMP-1 would be abnormal in hypertension reflecting alterations in E C M turnover. To test this hypothesis,
XIV bztentational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006
240
Conclusion: Hypoxia decreased the biosynthesis of cell associated HSPGs in macrophages. These changes may contribute to motility alterations leading to accumulation and clustering of macrophages in atherosclerotic lesions. Funding: ALF grants of SaJalgrenska University Hospital, the Heart and Lung Foundation, and AstraZeneca.
Tu-P7:251
P L A S M A M A R K E R S OF F I B R O S I S L E V E L S IN H Y P E R T E N S I O N IN C O M B I N A T I O N W I T H ISCHEMIC HEART DISEASE
A. Belovol, O. Kovalyova, E. Kolosov. State Medical Universi~, Kharkov,
Ukrabw Objective: We aimed to estimate the involvement of proform of matrix metallopro-teinase-1 (proMMP-1) and tissue inhibitor of metalloproteinases1 (TIMP-1) in left ventriculax (LV) remodeling in hypertension (HT) in combination with ischemic heart disease (IHD). Methods: 69 patients (pts) with HT in combination with IHD and 20 normotensive (NT) persons were examined. All pts were divided into 4 groups: normal geometry (NG), LV concentric remodeling (CR), LV concentric hypertrophy (CH) and LV ec-centric hypertrophy 0EH). Plasma proMMP-1 (TBS, United Kingdom) and TIMP-1 (BioSource International, Belgium) levels were determined with ELISA. Results: The plasma proMMP-1 level was higher in pts with HT with IHD than in NT persons (3,87-t-2,25 ng/ml versus 1,38-t-0,82 ng/ml, p<0,001). A direct correlation was found between plasma proMMP-1 level and LVMI (r--0,67, p<0,001), LV end dia-stolic-diameter index (r=0,66, p<0,001). Plasma TIMP-1 concentration was higher in pts with HT in combination with IHD than in NT persons (409,23+4,13 ng/ml versus 368,55+7,43 ng/ml, p<0,05). TIMP-1 level correlated with LVMI (r=0,38, p<0,05). Conclusions: Change of proMMP-1 level is compensatory mechanism in response to alteration of LV mass, size and myocardial structure. The excessive proMMP-1 production correlates with progressive increase of LVMI mainly relevant to LV chamber dimensions and less with degree LV wall thickness in patients with HT in combination with IHD. The high TIMP-1 level promotes increase of LVMI at the expense of collagen accumulation in myocardial extracellular matrix as a result of lowering activity of MMP-1.
Tu-P7:252
G R O U P IVA P H O S P H O L I P A S E A 2 - M E D I A T E D P R O D U C T I O N OF F I B R O N E C T I N BY O X I D I Z E D L D L IN M E S A N G I A L C E L L S
S. Akiba, Y. Mukaida, K. Hane, M. Oka, T. Sato. Kyoto Pharmaceutical
Universi~, Kyoto, Japan Objective: The development and progression of glomerulax diseases including glomerulosclerosis and diabetic nephropathy are associated with the accumulation of extracellular matrix proteins 0ECM) and the deposition of atherogenic lipoproteins such as low-density lipoprotein (LDL) and oxidized LDL (oxLDL) within the mesangium. The atherogenic lipoproteins are shown to stimulate the production of fibronectin and collagen, components of ECM, in mesangial cells. To clarify the mechanisms underlying the oxLDL-induced production of ECM, we examined the possible involvement of group IVA phospholipase A2 (PLA2) using human mesangial cells and group IVA PLA2-deficient mouse mesangial cells. Results: oxLDL accelerated the production of fibronectin and collagen (type IV) 6 h and 24 h after the stimulation, respectively. Under the conditions, oxLDL induced the release of axachidonic acid 3 h after the stimulation. Methyl arachidonyl fluorophosphonate (MAFP), an inhibitor of group IVA PLA2, markedly suppressed the oxLDL-induced production of fibronectin as well as the release of arachidonic acid, while it had no effect on the production of collagen. The inhibitory effect of MAFP on the production of fibronectin was reversed by adding arachidonic acid. Furthermore, in group IVA PLA2deficient mouse mesangial cells, the production of fibronectin induced by oxLDL was weak as compared with that in wild-type cells. However, the production by oxLDL of collagen was not affected in the deficient cells. Conclusion: These findings suggest that group IVA PLA2 is involved in the production of fibronectin in oxLDL-stimulated mesangial cells.
I Tu-P7:253 I S U B S T R A T E - D E P E N D E N T
R E G U L A T I O N OF
INFLAMMATORY
S. Filippov, M. Chambers, R. Homan. Esperion Therapeutics, Division of Pfizer Global Research and Development Ann Arbor; Michigan, USA In vulnerable atherosclerotic lesions, the dissolution of extracellulax matrix scaffolds is associated with the local accumulation of macrophages that express heightened levels of a diverse mix of pro-inflammatory cytokines and matrix-degrading enzymes. The regulation of monocyte differentiation by the nature and physical characteristics of the underlying substratum was examined in primary human monocytes that were adhered to either type I collagen or an uncoated plastic surface (representing physiological versus modified matrices, respectively). The protein expression profiles following monocyte to macrophage differentiation were assessed by antibody arrays. In addition to their very distinct shape and morphological appearance, macrophages differentiated on plastic expressed considerably higher levels of pro-inflammatory cytokines. Several of these, including IL-6, IL-8, MCP-1, MIP-lb, TNF-a and eotaxin-2, axe associated with lesion vulnerability in vivo. The macrophages exposed to the rigid surface of uncoated plastic consistently displayed marked upregulation of neutrophil collagenase (MMP-8), along with its most potent physiological activator stromelysin 2 (MMP- 10). Interestingly, the expression of other secreted members of matrix metalloproteinase family, MMP-1, MMP-2, MMP-3 and MMP-13, remained scarcely detectable under both culture conditions. Thus, adhesive interactions with the substratum direct human monocyte-macrophage differentiation and regulate macrophage tissue-destructive potential via expression of pro-inflammatory cytokines and the MMP-8/MMP- 10 axis.
Tu-P7:254
MATRIX METALLOPROTEINASES AND TISSUE I N H I B I T O R M E T A L L O P R O T E I N A S E S IN HYPERTENSIVE PATIENTS BEFORE AND AFTER ANTIHYPERTENSIVE THERAPY
G. Derosa 1 , S. Salvadeo 1 , A. D•Angelo 1 , C. Tinelli 2 , A.F.G. Cicero 3 , L. CiccaxelliI , M.N. Piccinni I , F. Pricolo I , A. Gravina I , R. Fogari I .
1Department of bttental Medicine attd Therapeutics, Universi~ of Pm'ia, Pcn'ia, Italy." 2Bionwtric Unit, IRCCS Policlinico San Matteo, Pavia, Italy." 3Departnwnt of Clinical Medicine attd Applied Biotechnology, Universi~ of Bologna, Bologna, Italy Aim: To test the hypothesis that MMP-2, MMP-9 and TIMP-1 axe altered in hypertension reflecting alterations in E C M turnover and to assess whether chronic antihypertensive treatment with doxazosin would normalize these alterations. M a t e r i a l s a n d M e t h o d s : We enrolled 44 hypertensive patients before and after 4 months treatment with doxazosin. We measured fasting plasma glucose (FPG), fasting plasma insulin (FPI), homeostasis model assessment index (HOMA index), systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), triglycerides (Tg), plasminogen activator inhibitor-1 (PAI-1), homocysteine (Hct) fibrinogen (Fg), C-reactive protein (hs-CRP), and plasma levels and activities of MMP-2, MMP-9 and TIMP- 1. Results: Significant FPI value decrease was observed in treated hypertensive group. SBP and DBP decrease was obtained in treated hypertensive group. Significant hs-CRP value was decreased in treated hypertensive patients. MMP-2 and MMP-9 levels and activity, and TIMP-1 were significantly lower in treated hypertensive group. Conclusions: Plasma levels and activities of MMP-2, MMP-9, and TIMP1 axe decreased in treated hypertensive patients compared to values before treatment.
ITu-P7:2551 MI NAHTIRB II XT OMREMT AELTLAOL PLROOP RT EOITNEAI SNEASS EASN DINT I S S U E HYPERTENSIVE PATIENTS •
l
.
.o
G. Derosa 1, A. D Angelo , C. Tinelh-, L. Ciccaxelli1 , M. Piccinni 1 , F. Pricolo I , S. Salvadeo I , M. Ghelfi I , I. Ferrari I , A. Cicero 3 . 1Department of
bttetTtal Medicine arm Therapeutics, Universi~ of Pavia, Pm,ia, Italy: 2Biometric Unit, IRCCS Policlinico San Matteo, Pavia, Italy." - Department of Clbtical Medicbw arm Applied Biotechnology, Universi~ of Bologna, Bologna, Italy Aim: We hypothesized that MMP-2, MMP-9 and TIMP-1 would be abnormal in hypertension reflecting alterations in E C M turnover. To test this hypothesis,
XIV bztentational Symposium on Atherosclerosis, Rome, Italy, June 18-22, 2006