Vascular smooth muscle cell phenotypic changes in patients with Marfan syndrome.

OBJECTIVE: Marfan's syndrome is characterized by the formation of ascending aortic aneurysms resulting from altered assembly of extracellular matrix microfibrils and chronic tissue growth factor (TGF)-ß signaling. TGF-ß is a potent regulator of the vascular smooth muscle cell (VSMC) phenotype. We hypothesized that as a result of the chronic TGF-ß signaling, VSMC would alter their basal differentiation phenotype, which could facilitate the formation of aneurysms. This study explores whether Marfan's syndrome entails phenotypic alterations of VSMC and possible mechanisms at the subcellular level. APPROACH AND RESULTS: Immunohistochemical and Western blotting analyses of dilated aortas from Marfan patients showed overexpression of contractile protein markers (α-smooth muscle actin, smoothelin, smooth muscle protein 22 alpha, and calponin-1) and collagen I in comparison with healthy aortas. VSMC explanted from Marfan aortic aneurysms showed increased in vitro expression of these phenotypic markers and also of myocardin, a transcription factor essential for VSMC-specific differentiation. These alterations were generally reduced after pharmacological inhibition of the TGF-ß pathway. Marfan VSMC in culture showed more robust actin stress fibers and enhanced RhoA-GTP levels, which was accompanied by increased focal adhesion components and higher nuclear localization of myosin-related transcription factor A. Marfan VSMC and extracellular matrix measured by atomic force microscopy were both stiffer than their respective controls. CONCLUSIONS: In Marfan VSMC, both in tissue and in culture, there are variable TGF-ß-dependent phenotypic changes affecting contractile proteins and collagen I, leading to greater cellular and extracellular matrix stiffness. Altogether, these alterations may contribute to the known aortic rigidity that precedes or accompanies Marfan's syndrome aneurysm formation.

Vasoactive effects of prostaglandins from the perivascular fat of mesenteric resistance arteries in WKY and SHROB rats.

AIMS: We have studied the vasoactive role of prostaglandins derived from perivascular adipose tissue (PVAT) and their effects on endothelial function in healthy rats and rats with metabolic syndrome (SHROB). MAIN METHODS: Mesenteric resistance arteries (MRA) from SHROB and control rats (WKY) were mounted on wire myographs: a) together with a sphere of naturally occurring perivascular adipose tissue (with-PVAT group), or b) dissecting all the adventitial tissue (without-PVAT group). KEY FINDINGS: Endothelial function, tested by acetylcholine reactivity of SHROB arteries with PVAT, was significantly lower than that of WKY. With-PVAT arteries, especially the SHROB, showed lower responses than those without PVAT. NO synthase inhibition diminished the acetylcholine responses in every group except the with-PVAT SHROB group. Blockade of cyclooxygenase-2, PGI2-IP, TXA2-TP, or TXA2 synthase increased to different extents the arterial responses in the SHROB with-PVAT group. PVAT from both rat strains revealed cyclooxygenase-2 activity and immunoassay confirmed the release of PGE2, PGI2 and TXA2. SIGNIFICANCE: Our major finding is that PVAT is a source of vasoactive prostaglandins in WKY and SHROB. We also report that the presence of visceral PVAT causes endothelial dysfunction of resistance arteries in the SHROB. The vascular responses to prostaglandins partly underlie the endothelial dysfunction of SHROB arteries.

Hypertension in metabolic syndrome: vascular pathophysiology.

METABOLIC SYNDROME IS A CLUSTER OF METABOLIC AND CARDIOVASCULAR SYMPTOMS: insulin resistance (IR), obesity, dyslipemia. Hypertension and vascular disorders are central to this syndrome. After a brief historical review, we discuss the role of sympathetic tone. Subsequently, we examine the link between endothelial dysfunction and IR. NO is involved in the insulin-elicited capillary vasodilatation. The insulin-signaling pathways causing NO release are different to the classical. There is a vasodilatory pathway with activation of NO synthase through Akt, and a vasoconstrictor pathway that involves the release of endothelin-1 via MAPK. IR is associated with an imbalance between both pathways in favour of the vasoconstrictor one. We also consider the link between hypertension and IR: the insulin hypothesis of hypertension. Next we discuss the importance of perivascular adipose tissue and the role of adipokines that possess vasoactive properties. Finally, animal models used in the study of vascular function of metabolic syndrome are reviewed. In particular, the Zucker fatty rat and the spontaneously hypertensive obese rat (SHROB). This one suffers macro- and microvascular malfunction due to a failure in the NO system and an abnormally high release of vasoconstrictor prostaglandins, all this alleviated with glitazones used for metabolic syndrome therapy.

Effects of pioglitazone and rosiglitazone on vascular function of mesenteric resistance arteries in rat genetic hypertension.

Glitazones exhibit beneficial effects in the vascular system, both on large vessels and at a microcirculatory level. We previously reported the effects of glitazones in the aorta of spontaneously hypertensive rats (SHR). We focus now on the acute and long-term actions of these drugs on mesenteric resistance arteries of the SHR. Incubation with pioglitazone or rosiglitazone (10⁻5 mol/l) improved endothelium-dependent relaxations to acetylcholine and the endothelial modulation of phenylephrine contractions. Acetylcholine relaxations that were abolished by N(G)-nitro-L-arginine methylester were partly recovered by the glitazones, but no effects of these drugs were observed in the presence of indomethacin or indomethacin + L-NAME. Glitazones did not change the contractions to U46619 or the endothelium-independent relaxation to sodium nitroprusside. Three-week oral pioglitazone or rosiglitazone treatment (3 and 10 mg/kg/day, respectively) confirmed the acute experiments. Thus, in microvessels, glitazones improve endothelial function in such a way that they do not alter endothelial nitric oxide release but reduce the production of vasoconstrictor prostanoids from endothelial cells.

Reactivity of the aorta and mesenteric resistance arteries from the obese spontaneously hypertensive rat: effects of glitazones.

The obese spontaneously hypertensive rat (SHROB) is a model of metabolic syndrome in which, to our knowledge, vascular function has never been studied. The actions of insulin sensitizers (glitazones) on vascular function have not been analyzed either. Our purpose was to characterize microvascular and macrovascular responses of the SHROB and to study the effects of glitazones on these responses. The reactivity of mesenteric resistance arteries (MRAs) and the aorta from SHROBs and control rats to cumulative concentrations of phenylephrine, ACh, and sodium nitroprusside (SNP) was myographically analyzed. Some animals were orally treated with rosiglitazone (3 mg·kg(-1)·day(-1), 3 wk), and myography was performed. Phenylephrine, ACh, and SNP dose-response curves were impaired to different extents in arteries of SHROBs. Incubation with N-nitro-L-arginine methyl ester caused little effects on phenylephrine and ACh curves in MRAs but enhanced phenylephrine contractions and abolished ACh-induced relaxations of aortae. Incubation with indomethacin reduced phenylephrine reactivity and improved ACh-induced relaxations of all vessels studied. NS-398 and tempol increased relaxations to ACh of MRAs. Incubation with pioglitazone or rosiglitazone (both 10(-5) M) or oral treatment with rosiglitazone improved, to different extents, ACh and SNP curves in all vessels. Glitazone incubation diminished aortic ACh sensitivity. The release of thromboxane A(2) and PGI(2) metabolites (thromboxane B(2) and 6-keto-PGF(1α)) was analyzed. ACh increased the MRA release of thromboxane B(2) from SHROBs but not control rats, and the former was prevented by rosiglitazone coincubation. In contrast, in aortae, ACh failed to alter the release of metabolites, and rosiglitazone treatment increased that of 6-keto-PGF(1α). Thus, SHROBs displayed microvascular and macrovascular dysfunction. MRAs, but not aortae, of SHROBs revealed an impaired endothelial nitric oxide pathway, whereas both, but especially MRAs, displayed an impaired cyclooxygenase pathway. Glitazones elicited beneficial effects on macrovascular and, especially, microvascular function of SHROBs.

Effects of pioglitazone and rosiglitazone on aortic vascular function in rat genetic hypertension.

Glitazones have beneficial antihypertensive effects independent of their insulin-sensitizing action. We have studied the effects of pioglitazone and rosiglitazone on the endothelial ability to counteract vascular smooth muscle contractility in genetic hypertension. To achieve this, we measured isometric responses of aortic segments obtained from spontaneously hypertensive rats. The effects of glitazones on endothelial function were studied by assessing the endothelial modulation of phenylephrine-induced isometric contractions (10(-9)-10(-5) M) in the presence or absence of pioglitazone or rosiglitazone (10(-5) M), added directly to an organ bath or orally administered to the rats (pioglitazone, 10 mg/kg). The role of both NO and prostanoids was analyzed by performing experiments in the presence of N(G)-nitro-L-arginine methyl ester (L-NAME) and/or indomethacin (both 10(-5) M) in the organ bath. Concentration-dependent contractions to L-NAME (10(-6)-3 x 10(-4) M) in the presence or absence of glitazones were carried out as an estimation of basal NO release. Pioglitazone, but not rosiglitazone, increased contractile responses to phenylephrine in intact vessels. The contractile responses to phenylephrine obtained in the presence of glitazones were markedly diminished by indomethacin, but enhanced by L-NAME. Analogous results were obtained in aortas from pioglitazone-chronically treated animals. L-NAME concentration-dependent contractions were enhanced by both glitazones. Both glitazones lowered the sensitivity to acetylcholine (10(-9)-10(-5) M). In conclusion, pioglitazone and rosiglitazone alter vascular function differentially and through endothelium-dependent mechanisms. These drugs act over the same pathways on the endothelium where they have a dual action, increasing both production of vasoconstrictor prostanoids and NO. The balance between both vasoactive substances determines the vascular response to glitazones.

The senescence-accelerated mouse (SAM-P8) as a model for the study of vascular functional alterations during aging.

We studied vascular function in quiescent aortas from senescence-accelerated resistant (SAM-R1) and prone (SAM-P8) mice. Myographical studies of thoracic aorta segments from 6-7 month-old mice showed that the contractility of SAM-P8 aortas was markedly higher than that of SAM-R1 after KCl depolarization or phenylephrine addition. Acetylcholine dose-response relaxation curves revealed that SAM-R1 vessels were slightly more sensitive than those of SAM-P8. In the presence of the NO synthase inhibitor, L-NAME, all vessels displayed contractions to acetylcholine, but these were more distinct in the SAM-R1. Phenylephrine plus L-NAME displayed stronger contractions in both animal strains, but were markedly more pronounced in SAM-R1. The cyclooxygenase inhibitor, indomethacin did not change the vessel responses to acetylcholine or phenylephrine. These data indicate that NO synthase, not cyclooxygenase, was responsible for the differences in contractility. Standard histology and immunohistochemistry of endothelial NO synthase revealed no differences in the expression of this protein. In contrast, increased levels of malondialdehyde were found in SAM-P8 vessels. We conclude that SAM-P8 vessels exhibit higher contractility than those of SAM-R1. Furthermore, our results suggest that the endothelium of SAM-P8 vessels is dysfunctional and lacks normal capability to counteract smooth muscle contraction. Therefore, our findings support SAM-P8 as a suitable model for the study of vascular physiological changes during aging.