- Email: [email protected]
122 Fenofibrate reduces apoptosis in endothelial cells in hyperglycemia
Abstracts X I X t h National Congress, Italian SocieO, f o r the Study o f A t h e l v s c l e l v s i s hyperlipidemia. Seven patient were treated, from at least twelve weeks, with stable doses of statins and they received E because they did not reach the NCEP goal of LDL. Nine patients did not received any statins and received only E therapy due to side effect of statins (6 have shown high levels of CPK and 3 of AST/ALT). All the patients have been evaluated after six weeks from the start of the treatment with E. A medical visit, dosage of TC, HDL, TG, LDL was performed before and six weeks after E therapy. The table shows the levels of lipid before and after E treatment. Ezetimibe
TC
HDL
TG
LDL
Before After 6 weeks Difference (%)
255.17±28.75 212.08±23.96 16.89
52.83±15.60 53.17±14.97 +0.4
176±90.62 138.5±73 21.31
169.83±18.38 132±22.13 22.27
Although the heterogeneity and the small number of subjects, its appears that E is effective in reduction of the lipid on hyperlipidemic outpatients. E has been supplied free of charge by Merck Sharp & Dohme.
F•
FENOFIBRATE REDUCES APOPTOSIS IN ENDOTHELIAL CELLS IN HYPERGLYCEMIA
A. Stocca, G. Grassi, B. Dapas, R. Farra, E Tedesco, L. Cattin, G. Guarnieri, M. ZaJnetti. (\linica Medica. Unic,ersit~'l di Trieste; Dipartimento di Fisiologia
e Patologia. [blic,ersitd di Trieste; UCO III Medica. Azienda Ospedaliera "Ospedali Riuniti" di Trieste. Italy E-mail: [email protected] Background: Cardiovascular disease is the leading cause of death in diabetic patients. Endothelial cell apoptosis, which is increased in hyperglycemia, is a proinflammatory and procoagulaaat condition predisposing to plaque rupture and thrombosis. PPAR,:~, (peroxisome-proliferator-activated-receptor alpha) agonists exert antiinflaanmatory and antiatherogenic effects on the vessel wall. In this study we hypothesized that increased apoptosis in hyperglycemia is prevented or reduced by a PPAR,:~, agonist. Methods: Human umbilical vein endothelial cells (HUVEC) were incubated in 5.5 or 22 mmol/l glucose with or without 50 ~tMol fenofibrate. In these conditions, the following determinations were carried out: cell proliferation by cell count and bromodeossiuridine incorporation test (BrdU), apoptosis by annexin V/FITC and Western blot analysis ofPARP (polyADP ribose polimerase), cytotoxicity by lactic-dehydrogenase (LDH). Results: at the tested concentration fenofibrate did not result in cytotoxicity (LDH test). In these experimental conditions reduced (P < 0.0005) cell proliferation in hyperglycemia (56±0.04% of cell counts and 91% of BrdU incorporation vs. control cells in 5.5 mmol/L glucose) was not prevented by coincubation with fenofibrate (62±0.06% and 52±0.01% respectively). However, fenofibrate reduced apoptosis in hyperglycemia by 40% as demonstrated by annexin V/FITC and Western Blot analysis for PARR Conclusions: Fenofibrate reduces endothelial cell apoptosis in hyperglycemia however does not prevent reduced proliferation in these conditions. EFFECTS OF ZOFENOPRIL ON ENDOTHELIUM-DEPENDENT F•-] VASODILATATION AND ON OXIDATIVE STRESS IN HYPERTENSIVE PATIENTS C. Stranieri, M.C. Nava, A. Fratta Pasini, U. Garbin, A. Davoli, ~ Lo Cascio, L. Cominacini. Department of Biomedical and Surgical Sciences (Medicina
D). University of ~'~rona. Italy E-mail: [email protected] Zofenopril is an ACE-inhibitor with, in vitro, antioxidant characteristic property, due to its SH group molecular structure. We studied 45 patients (15M, 20F) affected by essential mild-moderate hypertension not complicated, to value if Zofenopril could: 1) improve endothelial-dependent vasodilatation (restoring the NO availability); 2) decrease the oxidative stress (evaluated indirectly by measurement of peroxides, oxidized LDL (oxLDL) and some adhesion molecules). We conducted an open-label, randomized study with three parallel groups. Zofenopril 30mg (ACE inhibitor with SH group) was compared with Ramipril 2.5 mg (ACE inhibitor without SH group) and Atenolol 50 mg (13-blocker without any antioxidant activity). We evaluated: blood pressure (BP), endothelium dependent and independent vasodilatation by echocolordoppler at the brachial artery at baseline and after 8 weeks of therapy; moreover we measured some adhesion molecules (ICAM-1, VCAM-1 and E-selectin) by ELISA method and plasmatic peroxides by radioimmunologic method, at baseline and after 8 weeks. All the treatments determined a significant (p < 0.0001) reduction of blood pressure values whether systolic or diastolic. The treatment with Zofenopril caused a significant increased (p< 0.001) of endothelium-dependent vasodilatation and no modification in endothelium independent vasodilatation. Circulating levels of adhesion molecules, EDE peroxides, oxLDE were significantly reduced (respectively: p < 0.05, p < 0.0001,
$27
p < 0.05) only in patients treated with Zofenopril. Conclusions: Zofenopril seems to have, also in vivo, some antioxidant characteristic, reducing circulating levels of peroxides, oxLDL and adhesion molecules. Moreover it can increase the endothelium-dependent vasodilatation.
F•]
NONALCOHOLIC FATTY LIVER DISEASE INDEPENDENTLY PREDICTS INCIDENT CARDIOVASCULAR EVENTS IN TYPE 2 DIABETIC PATIENTS - FINDINGS FROM THE VALPOLICELLA HEART DIABETES STUDY
G. Taxgher, L. Bertolini, R. Tessaxi, L. Scala, L. Zenari, G. Falezza. Dic~ of Inte~Tzal medicine and Diabetes Unit. Sacro (~tore Hospital. Negrar (VR). Italy E-maih [email protected] Nonalcoholic fatty liver disease (NAFLD) is closely associated with several metabolic syndrome (MetS) features. We assessed prospectively whether NAFLD (as diagnosed by liver ultrasonography, blood testing and medical history) predicts future cardiovascular (CVD) events among type 2 diabetic individuals, independently of MetS components and other classical risk factors. We did a prospective, nested case-control, study in 2,103 type 2 diabetic patients, who were clinically free of diagnosed CVD at baseline. During 5 years of follow-up, 248 participants (cases) subsequently developed non-fatal coronary heart disease (myocardial infarction and coronary revascularization procedures) or ischaemic stroke or cardiovascular death. Using a risk set sampling, 496 patients (controls) among those who remained free of diagnosed CVD during follow-up were randomly selected in a 2:1 ratio, matched for age and sex to the cases. After adjustment for age, sex, smoking status, diabetes duration, hemoglobin Alc, LDL cholesterol, liver enzymes and use of medications, the presence of NAFLD was significantly associated with an increased CVD risk (odds ratio [OR], 1.84; 95%CI: 1.4-2.1; P < 0.001). Additional adjustment for the MetS (as defined by the Adult Treatment PaneMII criteria) appreciably attenuated, but did not abolish, this association (OR 1.53; 95%CI: 1.1-1.7; P 0.02). In conclusion, this is the first study demonstrating that NAFLD is associated with a moderately increased CVD risk among type 2 diabetic individuals. This relationship is independent of classical risk factors and is only partly explained by occurrence of MetS.
FI•
GHRELIN IMPROVES ENDOTHELIAL FUNCTION IN PATIENTS WITH METABOLIC SYNDROME
M. Tesauro, E Schinzari, M. IaJntorno, S. Rizza, D. Melina, D. Lauro, R. Lauro, C. Caxdillo. Department of Medicina Interna. Unic,ersitd di Tor ~'brgata. and
the Dic,isione di Terapia Medica Uomplesso hztegrato Uolumbus. Unic,ersitgl (~tttolica del Sacro (~tore. Rome. ItaO~ E-maih [email protected] Background: Metabolic syndrome (MetS) importantly accelerates the atherosclerotic process, the earliest event of which is endothelial dysfunction. Ghrelin, a gastric peptide with cardiovascular actions, has been shown to inhibit proatherogenic changes in experimental models. This study, therefore, investigatedthe possibility that ghrelin administration could beneficially affect endothelial function in MetS. Methods and Results: Endotheliumdependent and -independent vasodilator responsesto intraarterial infusion of acetylcholine (ACh; 7.5, 15, and30~tg/min) and sodium nitroprusside (SNP; 0.8, 1.6, and3.2 ~tg/min), respectively, were assessed by strain-gauge plethysmographybefore and after local administration of human ghrelin (200 ~tg/min). During saline,the vasodilator response to ACh was significantly blunted in patients with MetS compared to controls (P 0.008), whereasthe vasodilator effect of SNP was not different between groups(P 0.68). In patients with MetS, basal plasma ghrelin was significantly lower than in controls (P 0.02). In these patients, ghrelin infusion markedly increased intravascular concentrations of the peptide (P < 0.001) and resulted in a significant potentiation of thevasodilator response to ACh (P 0.001 vs. saline). This effect was associated to increased nitric oxide (NO) availability, as demonstrated by the significant decrease in blood flow response to ACh induced by N G-monomethylL-axginine following ghrelin administration (P 0.01). Conclusions: These findings indicate that ghrelin reverses endothelial dysfunction in patients with MetS by increasing NO bioactivity, thereby suggesting that decreased circulating levels of the peptide, such as those found in these patients, might play a role in the pathobiology of the atherosclerotic process.
TC
HDL
TG
LDL
Before After 6 weeks Difference (%)
255.17±28.75 212.08±23.96 16.89
52.83±15.60 53.17±14.97 +0.4
176±90.62 138.5±73 21.31
169.83±18.38 132±22.13 22.27
Although the heterogeneity and the small number of subjects, its appears that E is effective in reduction of the lipid on hyperlipidemic outpatients. E has been supplied free of charge by Merck Sharp & Dohme.
F•
FENOFIBRATE REDUCES APOPTOSIS IN ENDOTHELIAL CELLS IN HYPERGLYCEMIA
A. Stocca, G. Grassi, B. Dapas, R. Farra, E Tedesco, L. Cattin, G. Guarnieri, M. ZaJnetti. (\linica Medica. Unic,ersit~'l di Trieste; Dipartimento di Fisiologia
e Patologia. [blic,ersitd di Trieste; UCO III Medica. Azienda Ospedaliera "Ospedali Riuniti" di Trieste. Italy E-mail: [email protected] Background: Cardiovascular disease is the leading cause of death in diabetic patients. Endothelial cell apoptosis, which is increased in hyperglycemia, is a proinflammatory and procoagulaaat condition predisposing to plaque rupture and thrombosis. PPAR,:~, (peroxisome-proliferator-activated-receptor alpha) agonists exert antiinflaanmatory and antiatherogenic effects on the vessel wall. In this study we hypothesized that increased apoptosis in hyperglycemia is prevented or reduced by a PPAR,:~, agonist. Methods: Human umbilical vein endothelial cells (HUVEC) were incubated in 5.5 or 22 mmol/l glucose with or without 50 ~tMol fenofibrate. In these conditions, the following determinations were carried out: cell proliferation by cell count and bromodeossiuridine incorporation test (BrdU), apoptosis by annexin V/FITC and Western blot analysis ofPARP (polyADP ribose polimerase), cytotoxicity by lactic-dehydrogenase (LDH). Results: at the tested concentration fenofibrate did not result in cytotoxicity (LDH test). In these experimental conditions reduced (P < 0.0005) cell proliferation in hyperglycemia (56±0.04% of cell counts and 91% of BrdU incorporation vs. control cells in 5.5 mmol/L glucose) was not prevented by coincubation with fenofibrate (62±0.06% and 52±0.01% respectively). However, fenofibrate reduced apoptosis in hyperglycemia by 40% as demonstrated by annexin V/FITC and Western Blot analysis for PARR Conclusions: Fenofibrate reduces endothelial cell apoptosis in hyperglycemia however does not prevent reduced proliferation in these conditions. EFFECTS OF ZOFENOPRIL ON ENDOTHELIUM-DEPENDENT F•-] VASODILATATION AND ON OXIDATIVE STRESS IN HYPERTENSIVE PATIENTS C. Stranieri, M.C. Nava, A. Fratta Pasini, U. Garbin, A. Davoli, ~ Lo Cascio, L. Cominacini. Department of Biomedical and Surgical Sciences (Medicina
D). University of ~'~rona. Italy E-mail: [email protected] Zofenopril is an ACE-inhibitor with, in vitro, antioxidant characteristic property, due to its SH group molecular structure. We studied 45 patients (15M, 20F) affected by essential mild-moderate hypertension not complicated, to value if Zofenopril could: 1) improve endothelial-dependent vasodilatation (restoring the NO availability); 2) decrease the oxidative stress (evaluated indirectly by measurement of peroxides, oxidized LDL (oxLDL) and some adhesion molecules). We conducted an open-label, randomized study with three parallel groups. Zofenopril 30mg (ACE inhibitor with SH group) was compared with Ramipril 2.5 mg (ACE inhibitor without SH group) and Atenolol 50 mg (13-blocker without any antioxidant activity). We evaluated: blood pressure (BP), endothelium dependent and independent vasodilatation by echocolordoppler at the brachial artery at baseline and after 8 weeks of therapy; moreover we measured some adhesion molecules (ICAM-1, VCAM-1 and E-selectin) by ELISA method and plasmatic peroxides by radioimmunologic method, at baseline and after 8 weeks. All the treatments determined a significant (p < 0.0001) reduction of blood pressure values whether systolic or diastolic. The treatment with Zofenopril caused a significant increased (p< 0.001) of endothelium-dependent vasodilatation and no modification in endothelium independent vasodilatation. Circulating levels of adhesion molecules, EDE peroxides, oxLDE were significantly reduced (respectively: p < 0.05, p < 0.0001,
$27
p < 0.05) only in patients treated with Zofenopril. Conclusions: Zofenopril seems to have, also in vivo, some antioxidant characteristic, reducing circulating levels of peroxides, oxLDL and adhesion molecules. Moreover it can increase the endothelium-dependent vasodilatation.
F•]
NONALCOHOLIC FATTY LIVER DISEASE INDEPENDENTLY PREDICTS INCIDENT CARDIOVASCULAR EVENTS IN TYPE 2 DIABETIC PATIENTS - FINDINGS FROM THE VALPOLICELLA HEART DIABETES STUDY
G. Taxgher, L. Bertolini, R. Tessaxi, L. Scala, L. Zenari, G. Falezza. Dic~ of Inte~Tzal medicine and Diabetes Unit. Sacro (~tore Hospital. Negrar (VR). Italy E-maih [email protected] Nonalcoholic fatty liver disease (NAFLD) is closely associated with several metabolic syndrome (MetS) features. We assessed prospectively whether NAFLD (as diagnosed by liver ultrasonography, blood testing and medical history) predicts future cardiovascular (CVD) events among type 2 diabetic individuals, independently of MetS components and other classical risk factors. We did a prospective, nested case-control, study in 2,103 type 2 diabetic patients, who were clinically free of diagnosed CVD at baseline. During 5 years of follow-up, 248 participants (cases) subsequently developed non-fatal coronary heart disease (myocardial infarction and coronary revascularization procedures) or ischaemic stroke or cardiovascular death. Using a risk set sampling, 496 patients (controls) among those who remained free of diagnosed CVD during follow-up were randomly selected in a 2:1 ratio, matched for age and sex to the cases. After adjustment for age, sex, smoking status, diabetes duration, hemoglobin Alc, LDL cholesterol, liver enzymes and use of medications, the presence of NAFLD was significantly associated with an increased CVD risk (odds ratio [OR], 1.84; 95%CI: 1.4-2.1; P < 0.001). Additional adjustment for the MetS (as defined by the Adult Treatment PaneMII criteria) appreciably attenuated, but did not abolish, this association (OR 1.53; 95%CI: 1.1-1.7; P 0.02). In conclusion, this is the first study demonstrating that NAFLD is associated with a moderately increased CVD risk among type 2 diabetic individuals. This relationship is independent of classical risk factors and is only partly explained by occurrence of MetS.
FI•
GHRELIN IMPROVES ENDOTHELIAL FUNCTION IN PATIENTS WITH METABOLIC SYNDROME
M. Tesauro, E Schinzari, M. IaJntorno, S. Rizza, D. Melina, D. Lauro, R. Lauro, C. Caxdillo. Department of Medicina Interna. Unic,ersitd di Tor ~'brgata. and
the Dic,isione di Terapia Medica Uomplesso hztegrato Uolumbus. Unic,ersitgl (~tttolica del Sacro (~tore. Rome. ItaO~ E-maih [email protected] Background: Metabolic syndrome (MetS) importantly accelerates the atherosclerotic process, the earliest event of which is endothelial dysfunction. Ghrelin, a gastric peptide with cardiovascular actions, has been shown to inhibit proatherogenic changes in experimental models. This study, therefore, investigatedthe possibility that ghrelin administration could beneficially affect endothelial function in MetS. Methods and Results: Endotheliumdependent and -independent vasodilator responsesto intraarterial infusion of acetylcholine (ACh; 7.5, 15, and30~tg/min) and sodium nitroprusside (SNP; 0.8, 1.6, and3.2 ~tg/min), respectively, were assessed by strain-gauge plethysmographybefore and after local administration of human ghrelin (200 ~tg/min). During saline,the vasodilator response to ACh was significantly blunted in patients with MetS compared to controls (P 0.008), whereasthe vasodilator effect of SNP was not different between groups(P 0.68). In patients with MetS, basal plasma ghrelin was significantly lower than in controls (P 0.02). In these patients, ghrelin infusion markedly increased intravascular concentrations of the peptide (P < 0.001) and resulted in a significant potentiation of thevasodilator response to ACh (P 0.001 vs. saline). This effect was associated to increased nitric oxide (NO) availability, as demonstrated by the significant decrease in blood flow response to ACh induced by N G-monomethylL-axginine following ghrelin administration (P 0.01). Conclusions: These findings indicate that ghrelin reverses endothelial dysfunction in patients with MetS by increasing NO bioactivity, thereby suggesting that decreased circulating levels of the peptide, such as those found in these patients, might play a role in the pathobiology of the atherosclerotic process.