The relationship among antioxidant activity, based on the electron-spin resonance determination of the reduction of Fremy's radical, vasodilation activity, and phenolic content was investigated in 16 red wines. The wines were selected to provide a range of origins, grape varieties, and vinification methods. Sensitive and selective HPLC methods were used for the analysis of the major phenolics in red wine: free and conjugated myricetin, quercetin, kaempferol, and isorhamnetin; (+)-catechin, (-)-epicatechin, gallic acid, p-coumaric acid, caffeic acid, caftaric acid, trans-resveratrol, cis-resveratrol, and trans-resveratrol glucoside. Total anthocyanins were measured using a colorimetric assay. The total phenolic content of the wines was determined according to the Folin-Ciocalteu colorimetric assay and also by the cumulative measurements obtained by HPLC. The 16 wines exhibited a wide range in the values of all parameters investigated. However, the total phenol contents, measured both by HPLC and colorimetrically, correlated very strongly with the antioxidant activity and vasodilation activity. In addition, the antioxidant activity was associated with gallic acid, total resveratrol, and total catechin. In contrast, only the total anthocyanins were correlated with vasodilation activity. The results demonstrate that the different phenolic profiles of wines can produce varying antioxidant and vasodilatant activities, which opens up the possibility that some red wines may provide enhanced health benefits for the consumer.

Background-Pulmonary arterial hypertension (PAH) is a hyperproliferative vascular disorder observed predominantly in women. Estrogen is a potent mitogen in human pulmonary artery smooth muscle cells and contributes to PAH in vivo; however, the mechanisms attributed to this causation remain obscure. Curiously, heightened expression of the estrogenmetabolizing enzyme cytochrome P450 1B1 (CYP1B1) is reported in idiopathic PAH and murine models of PAH. Methods and Results-Here, we investigated the putative pathogenic role of CYP1B1 in PAH. Quantitative reverse transcriptionpolymerase chain reaction, immunoblotting, and in situ analysis revealed that pulmonary CYP1B1 is increased in hypoxic PAH, hypoxicϩSU5416 PAH, and human PAH and is highly expressed within the pulmonary vascular wall. PAH was assessed in mice via measurement of right ventricular hypertrophy, pulmonary vascular remodeling, and right ventricular systolic pressure. Hypoxic PAH was attenuated in CYP1B1 Ϫ/Ϫ mice, and the potent CYP1B1 inhibitor 2,3Ј,4,5Ј-tetramethoxystilbene (TMS; 3 mg ⅐ kg Ϫ1 ⅐ d

Abstract-Heterozygous germline mutations in the gene encoding the bone morphogenetic protein type II (BMPR-II) receptor underlie the majority (Ͼ70%) of cases of familial pulmonary arterial hypertension (FPAH), and dysfunction of BMP signaling has been implicated in other forms of PAH. The reduced disease gene penetrance in FPAH indicates that other genetic and/or environmental factors may also be required for the clinical manifestation of disease. Of these, the serotonin pathway has been implicated as a major factor in PAH pathogenesis. We investigated the pulmonary circulation of mice deficient in BMPR-II (BMPR2 ϩ/Ϫ mice) and show that pulmonary hemodynamics and vascular morphometry of BMPR2 ϩ/Ϫ mice were similar to wild-type littermate controls under normoxic or chronic hypoxic (2-to 3-week) conditions. However, chronic infusion of serotonin caused increased pulmonary artery systolic pressure, right ventricular hypertrophy, and pulmonary artery remodeling in BMPR2 ϩ/Ϫ mice compared with wild-type littermates, an effect that was exaggerated under hypoxic conditions. In addition, pulmonary, but not systemic, resistance arteries from BMPR2 ϩ/Ϫ mice exhibited increased contractile responses to serotonin mediated by both 5-HT 2 and 5-HT 1 receptors. Furthermore, pulmonary artery smooth muscle cells from BMPR2 ϩ/Ϫ mice exhibited a heightened DNA synthesis and activation of extracellular signal-regulated kinase 1/2 in response to serotonin compared with wild-type cells. In vitro and in vivo experiments suggested that serotonin inhibits BMP signaling via Smad proteins and the expression of BMP responsive genes. These findings provide the first evidence for an interaction between BMPR-II-mediated signaling and the serotonin pathway, perturbation of which may be critical to the pathogenesis of PAH. Key Words: pulmonary arterial hypertension Ⅲ bone morphogenetic protein Ⅲ serotonin I diopathic pulmonary arterial hypertension (IPAH) is characterized by narrowing and obliteration of the small arteries of the lung leading to increased pulmonary vascular resistance. 1 Elevation of pulmonary arterial pressure leads to right ventricular failure. Patients present with dyspnea after exertion, and many died of right heart failure within 3 years of diagnosis, before modern therapies. 2 Treatment with prostanoids or endothelin receptor antagonists improves symptoms and survival, although, for many, the long-term outlook remains poor.Heterozygous germline mutations in the gene encoding the bone morphogenetic protein type II receptor (BMPR-II) occur in up to 70% of cases of familial PAH (FPAH). 3,4 Similar mutations were found in up to 26% of cases of idiopathic PAH. 5 However, the low disease gene penetrance suggests that other genetic or environmental factors are necessary to manifest clinical disease.A series of studies have implicated serotonin (or 5-HT) as a key mediator of PAH. 6 -8 For example, treatment of rats with serotonin potentiates the effects of hypoxia on pulmonary arterial pressure, right ventricular hypertrophy an...

1 We investigated 5-hydroxytryptamine (5-HT)-receptor mediated vasoconstriction in the main, first branch and resistance pulmonary arteries removed from control and pulmonary hypertensive rats. Contractile responses to 5-HT, 5-carboxamidotryptamine (5-CT, non-selective 5-HT, agonist), and sumatriptan (5-HTID-like receptor agonist) were studied. The effects of methiothepin (non-selective 5-HT, ±2-receptor antagonist) and ketanserin (5-HT2A receptor antagonist) and GR55562 (a novel selective 5-HTID receptor antagonist) on 5-HT-mediated responses were also studied. Basal levels of adenosine 3':5'-cycic monophosphate ([cyclic AMP]j) and guanosine 3':5'-cyclic monophosphate ([cyclic GMP]j) were determined and we assessed the degree of inherent tone in the vessels under study.2 5-HT was most potent in the resistance arteries. pEC50 values were 5.6 + 0.1, 5.3 + 0.1, 5.0 + 0.2 in the resistance arteries, pulmonary branch and main pulmonary artery, respectively (n = at least 5 from 5 animals). The sensitivity to, and maximum response of, 5-HT was increased in all the arteries removed from the chronic hypoxic (CH) rats. In CH rats the pEC50 values were 5.9 + 0.2, 6.3 +0.2, 6.4 + 0.2 and the increase in the maximum response was 35%, 51% and 41% in the resistance arteries, pulmonary branch and main pulmonary artery, respectively. Sumatriptan did not contract any vessel from the control rats whilst 5-CT did contract the resistance arteries. In the CH rats, however, they both contracted the resistance arteries (responses to sumatriptan were small) (pEC50: 5-CT; 5.4+0.2) and the pulmonary artery branches (pEC50: sumatriptan, 5.4 +0.2; 5-CT, 5.4 +0.2). 5-CT also caused a contraction in the main pulmonary artery (pECmo: 6.0+0.3). 3 Ketanserin (1 nM-l gM) caused a competitive antagonism of the 5-HT response in all vessels tested.In control rats, the estimated pKb values for ketanserin in resistance arteries, pulmonary branches and main pulmonary artery were 8.3, 7.8 and 9.2, respectively. Methiothepin (1 nM-l giM) inhibited responses to 5-HT in the first branch (estimated pKb value: 7.8) and main pulmonary artery. In CH rats, the estimated pKb values for ketanserin in resistance arteries, pulmonary branches and main pulmonary artery were 7.7, 8.3 and 9.6, respectively. Methiothepin also inhibited contractions to 5-HT in the pulmonary artery branch and main pulmonary artery with estimated pKb values of 7 and 9.5, respectively. In control animals, GR55562 had no effect on responses to 5-HT in any of the vessels tested. In the CH rats the estimated pKb values for GR55562 were 6.5, 7.8 and 7.0 in the pulmonary resistance arteries, first branch and main pulmonary artery, respectively. 6 The results suggest that the increased vasoconstrictor response to 5-HT in CH rat pulmonary arteries is due to an increase in 5-HT2A-receptor mediated contraction combined with an increase in r5-HTIB-like receptor-mediated contraction. An increase in vascular tone and decreased levels of [cyclic GMP]i in the large pulmonary arteries may contribute to...

Responses to human urotensin-II (hU-II) were investigated in human and rat pulmonary arteries. Rat pulmonary arteries: hU-II was a potent vasoconstrictor of main pulmonary arteries (2 ± 3 mm i.d.) (pEC 50 , 8.55+0.08, n=21) and was *4 fold more potent than endothelin-1 [ET-1] (P50.01), although its E max was considerably less (*2.5 fold, P50.001). The potency of hU-II increased 2.5 fold with endothelium removal (P50.05) and after raising vascular tone with ET-1 (P50.01). E max was enhanced *1.5 fold in the presence of N o -nitro-L-arginine methylester (L-NAME, 100 mM, P50.01) and *2 fold in vessels from pulmonary hypertensive rats exposed to 2 weeks chronic hypoxia (P50.05). hU-II did not constrict smaller pulmonary arteries. Human pulmonary arteries (*250 mm i.d.): in the presence of L-NAME, 3 out of 10 vessels contracted to hU-II and this contraction was highly variable. hU-II is, therefore, a potent vasoconstrictor of rat main pulmonary arteries and this response is increased by endothelial factors, vascular tone and onset of pulmonary hypertension. Inhibition of nitric oxide synthase uncovers contractile responses to hU-II in human pulmonary arteries.

1 The 5-hydroxytryptamine (5-HT) receptors mediating vasoconstriction in isolated human small muscular pulmonary arteries (SMPAs) were determined using techniques of wire myography and reverse transcription-polymerase chain reaction (RT ± PCR). 2 The agonists 5-HT, 5-carboxamidotryptamine (5-CT, unselective for 5-HT 1 receptors) and sumatriptan (selective for 5-HT 1B/D receptors) all caused contraction and were equipotent (pEC 50 s: 7.0+0.2, 7.1+0.3 and 6.7+0.1, respectively) suggesting the presence of a 5-HT 1 receptor. 3 Ketanserin (5-HT 2A -selective antagonist, 0.1 mM) inhibited 5-HT-induced contractions only at non-physiological/pathological concentrations of 5-HT (40.1 mM) whilst GR55562 (5-HT 1B/1D -selective antagonist, 1 mM) inhibited 5-HT-induced contractions at all concentrations of 5-HT (estimated pK B =7.7+0.2). SB-224289 (5-HT 1B -selective antagonist, 0.2 mM) inhibited sumatriptaninduced contractions (estimated pK B =8.4+0.1) whilst these were una ected by the 5-HT 1D -selective antagonist BRL15572 (0.5 mM) suggesting that the 5-HT 1B receptor mediates vasoconstriction in this vessel. 4 RT ± PCR con®rmed the presence of substantial amounts of mRNA for the 5-HT 2A and 5-HT 1B receptor subtypes in these arteries whilst only trace amounts of 5-HT 1D receptor message were evident. 5 These ®ndings suggest that a heterogeneous population of 5-HT 2A and 5-HT 1B receptors co-exist in human small muscular pulmonary arteries but that the 5-HT 1B receptor mediates 5-HT-induced vasoconstriction at physiological and pathophysiological concentrations of 5-HT. These results have important implications for the treatment of pulmonary hypertension in which the 5-HT 1B receptor may provide a novel and potentially important therapeutic target.

Abstract-Tryptophan hydroxylase 1 catalyzes the rate-limiting step in the synthesis of serotonin in the periphery.Recently, it has been shown that expression of the tryptophan hydroxylase 1 gene is increased in lungs and pulmonary endothelial cells from patients with idiopathic pulmonary arterial hypertension. Here we investigated the effect of genetic deletion of tryptophan hydroxylase 1 on hypoxia-induced pulmonary arterial hypertension in mice by measuring pulmonary hemodynamics and pulmonary vascular remodeling before and after 2 weeks of hypoxia. In wild-type mice, hypoxia increased right ventricular pressure and pulmonary vascular remodeling. These effects of hypoxia were attenuated in the tryptophan hydroxylase 1 Ϫ/Ϫ mice. Hypoxia increased right ventricular hypertrophy in both wild-type and tryptophan hydroxylase 1 Ϫ/Ϫ mice suggesting that in vivo peripheral serotonin has a differential effect on the pulmonary vasculature and right ventricular hypertrophy. Contractile responses to serotonin were increased in pulmonary arteries from tryptophan hydroxylase 1 Ϫ/Ϫ mice. Hypoxia increased serotonin-mediated contraction in vessels from the wild-type mice, but this was not further increased by hypoxia in the tryptophan hydroxylase 1 Ϫ/Ϫ mice. In conclusion, these results indicate that tryptophan hydroxylase 1 and peripheral serotonin play an essential role in the development of hypoxia-induced elevations in pulmonary pressures and hypoxia-induced pulmonary vascular remodeling. In addition, the results suggest that, in mice, serotonin has differential effects on the pulmonary vasculature and right ventricular hypertrophy. Serotonin promotes pulmonary arterial smooth muscle cell proliferation, pulmonary arterial vasoconstriction, and local microthrombosis. 4 Proliferation of pulmonary arterial smooth muscle cells is an important component of pulmonary arterial remodeling in PAH, which accounts for the increased thickness of the medial muscular coat in normally muscularized arteries and extension of muscle into smaller and more peripheral arteries. Elevated circulating levels of peripheral serotonin have been associated with the development of PAH clinically. 5 It has also been shown that exogenously administered serotonin can potentiate the development of PAH in rats 6 and can uncover a PAH phenotype in bone morphogenetic protein receptor II ϩ/Ϫ mice. 7 Moreover, mice overexpressing SERT (SERTϩ mice) develop spontaneous PAH and are more susceptible to hypoxia-induced PAH, whereas mice deficient for the SERT are less susceptible. 8 -10 Blood serotonin levels are also elevated in mice after hypoxic exposure. 11 Tryptophan hydroxylase (Tph) catalyzes the rate-limiting step in the synthesis of serotonin from tryptophan. By studying Tph1 Ϫ/Ϫ

Pulmonary arterial 5-hydroxytryptamine (serotonin) (5-HT) transporter (SERT)-, 5-HT receptor expression, and 5-HTinduced vasoconstriction can be increased in pulmonary hypertension. These variables were studied in normoxic and hypoxic Fawn-Hooded (FH) and Sprague-Dawley (SD) rats. Furthermore, we compared the functional effects of SERT inhibitors and 5-HT receptor antagonists against 5-HTinduced vasoconstriction of pulmonary arteries. SERT and 5-HT 1B expression was greater in FH rat lungs than in SD rats, as was 5-HT-mediated vasoconstriction. However, these inhibitors potentiated responses to 5-HT in FH vessels. After exposure of rats to 2 weeks of hypoxia, there was increased 5-HTmediated vasoconstriction and a profound decrease in SERT expression in both the FH and SD rat lung. Accordingly, citalopram had no effect on 5-HT-induced constriction in SD rat vessels and markedly less effect in FH rat vessels. Ketanserin, SB224289, and LY393558 inhibited responses to 5-HT in all hypoxic rat vessels. LY393558 was the most potent antagonist, and there was synergy between the effects of fluoxetine and SB224289 when given simultaneously. The results suggest that, in FH rats, SERT inhibitors may increase pulmonary vasoconstriction, but this can be inhibited by simultaneous 5-HT 1B receptor antagonism. There is synergy between the inhibitory effects of 5-HT 1B receptor antagonists and SERT inhibitors on 5-HT-induced pulmonary vasoconstriction.Pulmonary arterial hypertension (PAH) is characterized by sustained elevation in pulmonary artery pressure. Familial PAH can be related to heterozygous germline mutations in the gene encoding the bone morphogenetic protein type II receptor and/or polymorphisms in the gene encoding the 5-hydroxytryptamine (serotonin) (5-HT) transporter (SERT) (Lane et al., 2000;Eddahibi et al., 2001). Idiopathic PAH has no demonstrable cause and PAH can also occur secondary to many cardiorespiratory disorders. Regardless of the type of PAH, the elevated pulmonary vascular resistance is associated with remodeling of muscular pulmonary arteries and arterioles that exhibit smooth muscle proliferation, medial hypertrophy, and fibrosis (Fishman, 1998).SERT mRNA is elevated in platelets from patients with PAH (Eddahibi et al., 2001). A polymorphism with long and short forms (Lesch et al., 1996) affects SERT function with the long allele inducing an increased rate of SERT gene transcription. The SERT polymorphism can also predict the severity of PAH in patients with chronic obstructive pulmonary disease (Eddahibi et al., 2003). Hence, it has been hypothesized that inhibitors of SERT may be useful in the treatment of PAH. One mechanism of action of SERT inhibitors treating clinical depression, however, is to cause an extracellular accumulation of 5-HT and increased 5-HT receptor activation (Slattery et al., 2004). The possibility that Article, publication date, and citation information can be found at