Huy Hao Dao

A new model of isolated systolic hypertension induced by chronic warfarin and vitamin K1 treatment

BACKGROUND Isolated systolic hypertension is the predominant form of hypertension in the elderly population. Reduction of arterial compliance appears to contribute to the elevation of pulse pressure (PP) and among potential mechanisms, gradual vascular calcification, fragmentation of elastic lamellae, and augmentation of rigid component like collagen could contribute to increase aortic stiffening. Few experimental models of the disease are currently available. METHODS To induce large artery calcification, rats were treated with warfarin and vitamin K(1) (WK) for 4 and 8 weeks, to inhibit the maturation of matrix Gla protein. The impact of chronic PP elevation was determined on large artery and cardiac remodeling and on aortic endothelial function. RESULTS The WK treatment led to aortic medial calcification and a proportional elevation of PP, attributable mainly to a selective elevation of systolic blood pressure. The chronic treatment also increased collagen, whereas elastin decreased in the aorta. Pulse wave velocity, an index of aortic stiffening, increased in rats treated with WK. However, indices of left ventricular and aortic hypertrophy and remodeling remained normal. In addition, the WK treatment did not modify the vasoconstriction to norepinephrine and endothelin-1, and the vasodilatory response to acetylcholine and sodium nitroprusside. CONCLUSIONS Chronic treatment with WK represents a new model of isolated systolic hypertension with several characteristics of the human disease. The relative ease to induce calcification in this model may help to foster more fundamental research, which is lacking in this type of hypertension.

Pharmacological prevention and regression of arterial remodeling in a rat model of isolated systolic hypertension.

Objectives and design Isolated systolic hypertension (ISH) is the predominant form of hypertension in the elderly population and drug treatment is unsatisfactory. We compared the efficacy of an endothelin-receptor antagonist (darusentan), an angiotensin-receptor blocker (irbesartan) and a thiazide diuretic (hydrochlorothiazide, HCTZ) to prevent and regress pulse pressure (PP) elevation and remodeling of large and small arteries, in a rat model of ISH obtained by the chronic administration of warfarin and vitamin K1 (WK). Methods and results Warfarin and vitamin K1 treatment for 4 or 8 weeks led to an elevation of PP, associated with increases in aortic calcium deposition and the ratio of collagen to elastin (C/E). Despite these changes in the composition of the aortic wall, the global structure of the aorta was unchanged. In contrast, an outward hypertrophic remodeling was observed in the middle cerebral artery. An early treatment with all drugs (darusentan, irbesartan, HCTZ) prevented PP elevation, changes of aortic media composition and the development of vascular remodeling. However, after 4 weeks of ISH, only darusentan and irbesartan reduced PP when administered from week 4 to 8. Darusentan was the most effective to regress existent aortic calcification, while only irbesartan reversed small artery hypertrophic remodeling. Conclusions During the development of ISH, drug treatment appears more beneficial when started early. Indeed, the three agents prevented PP elevation, aortic calcification and C/E increase in the aorta, and hypertrophy in small arteries. In contrast, once the disease is established, endothelin appears crucial in the maintenance of aortic calcification, while angiotensin II sustains small artery hypertrophy.

Norepinephrine-induced aortic hyperplasia and extracellular matrix deposition are endothelin-dependent.

Background Sympathetic hyperactivity is observed in several disease states and may contribute to cardiovascular hypertrophic remodeling. Endothelin has been suggested to be a mediator of hypertrophy. Objective To examine the involvement of endothelin in maintaining the growth response induced by exogenous norepinephrine. Design and methods Rats were treated with norepinephrine (2.5 μg/Kg per min subcutaneously) for 2 and 4 weeks, alone or in association with the selective endothelin-A (ETA) receptor antagonist, darusentan (LU135252, 30 mg/Kg per day orally) for weeks 3 and 4. Results Increases in medial cell number and accumulation of collagen and elastin characterized norepinephrine-induced aortic remodeling. These effects occurred without marked changes of mean arterial pressure, but may be related to enhanced pressure variability in addition to direct effects of norepinephrine. Inhibition of ETA receptors by darusentan reversed aortic alterations produced by infusion of norepinephrine. Evaluation of medial apoptosis did not reveal any significant change in any group at 4 weeks. Conclusions Antagonism of ETA receptors effectively and rapidly reversed norepinephrine-induced aortic structural and compositional changes, suggesting a central role of endothelin in mediating this response. Thus, ETA receptor antagonists may help to regress large artery remodeling in conditions of increased circulating catecholamine concentrations.

Transient involvement of endothelin in hypertrophic remodeling of small arteries.

Objective: This study was designed to evaluate the capacity of norepinephrine (NE) to induce hypertrophic remodeling of small arteries in rats, and to determine the involvement of endothelin (ET) to initiate and maintain it. Design and results: Treatment with NE (2.5 μ g/kg per min) for 14 or 28 days produced a similar inward hypertrophic remodeling, characterized by a smaller lumen, but increased media thickness and cross-sectional area. Arterial stiffness was reduced. Histological evaluation confirmed the hypertrophic nature of remodeling. Concomitant administration of LU135252 (ET-receptor antagonist) for the first 14 days of NE administration prevented the development of hypertrophy, without altering arterial mechanics. Treatment with the same antagonist from day 14 to day 28 of NE or angiotensin II (Ang II) treatment failed to regress established vascular hypertrophy. In contrast, normalization of arterial structure was observed with prazosin, an α-adrenergic blocker. Endothelin content in small mesenteric arteries showed a transient elevation following chronic NE administration. Conclusions: Increased circulating NE levels are associated with hypertrophic remodeling of small arteries, in which ET plays an initiating role. However, the maintenance of vascular hypertrophy is ET-independent, either in the presence of augmented circulating levels of NE or Ang II. Thus, early rather than late treatment with ET-receptor antagonists may be a preferable approach to limit small artery-mediated end-organ damage in cardiovascular diseases.

Regression of Medial Elastocalcinosis in Rat Aorta A New Vascular Function for Carbonic Anhydrase

Background—We sought to determine whether carbonic anhydrase (CA), which plays an important role in bone resorption, contributes to vascular mineral loss induced by an endothelin receptor antagonist. Methods and Results—Wistar rats were compared with rats receiving warfarin and vitamin K1 (WVK) for 8 weeks alone or in association with the endothelin receptor antagonist darusentan (30 mg/kg per day), the CA inhibitor acetazolamide (100 mg/kg per day), or both for the last 4 weeks. Rats were also treated with WVK for 5 or 6 weeks, and darusentan was added for the last week or last 2 weeks of treatment, respectively. Treatment with WVK produced medial elastocalcinosis in the aorta and carotid arteries. Immunohistochemistry revealed that CA II was already abundant in the adventitia and in calcified areas of aortic sections from WVK-treated rats. Darusentan did not significantly modify its abundance or distribution. In contrast, CA IV immunostaining, which was weak in WVK-treated rats, became apparent after 1 week of darusentan treatment and declined toward basal levels thereafter. These findings were confirmed by a parallel increase in CA IV protein abundance and activity in the aorta. The mineral loss induced by darusentan was blunted by acetazolamide treatment, confirming the functional relevance of the biochemical findings. Moreover, CA IV immunostaining was enhanced much later in the carotids, where darusentan did not cause regression of elastocalcinosis. Conclusions—Vascular mineral loss induced by the blockade of endothelin receptors seems dependent on the activation of membrane-bound CA IV, suggesting that mineral loss may proceed via local changes in pH similar to that seen in bone resorption.

Phenotypic modulation of vascular smooth muscle cells during medial arterial calcification: a role for endothelin?

We have previously shown that an endothelin receptor antagonist can regress medial arterial calcification in a rat model. The aim of this study was to characterize the phenotypic changes of vascular smooth muscle cells during calcification and mineral loss, in order to understand better the underlying mechanisms. Control Wistar rats were compared with rats treated only with warfarin/ vitamin K1 (15 mg/kg per day) for 8 weeks, or in combination with darusentan (30 mg/kg per day) for the final 4 weeks. Vascular smooth muscle cell, bone cell and macrophage phenotypes were evaluated by the local expression of α-actin, tartrate-resistant acid phosphatase and ED-1, respectively. Proteins involved in the modulation of bone resorption like osteopontin and osteoprotegerin were also evaluated by immunohistochemistry. The warfarin/vitamin K1 treatment increased medial arterial calcification ninefold (P < 0.05). At sites of calcification, there was a decrease in α-actin localization, and an appearance of osteopontin immunostaining. Histochemical and immunostaining for osteoclast and macrophage markers, as well as for osteoprotegerin, were negative. Although the extent of calcification foci was reduced by darusentan, protein localization in the calcified areas was not modified. Thus, the development of medial arterial calcification produces a phenotypic change in vascular smooth muscle cells that does not appear to be normalized in regions remaining calcified during mineral loss.

Signaling of angiotensin II-induced vascular protein synthesis in conduit and resistance arteries in vivo

BackgroundFrom in vitro studies, it has become clear that several signaling cascades are involved in angiotensin II-induced cellular hypertrophy. The aim of the present study was to determine some of the signaling pathways mediating angiotensin II (Ang II)-induced protein synthesis in vivo in large and small arteries.MethodsNewly synthesized proteins were labeled during 4 hours with tritiated leucine in conscious control animals, or animals infused for 24 hours with angiotensin II (400 ng/kg/min). Hemodynamic parameters were measure simultaneously. Pharmacological agents affecting signaling cascades were injected 5 hours before the end of Ang II infusion.ResultsAngiotensin II nearly doubled the protein synthesis rate in the aorta and small mesenteric arteries, without affecting arterial pressure. The AT1 receptor antagonist Irbesartan antagonized the actions of Ang II. The Ang II-induced protein synthesis was associated with increased extracellular signal-regulated kinases (ERK)1/2 phosphorylation in aortic, but not in mesenteric vessels. Systemic administration of PD98059, an inhibitor of the ERK-1/2 pathway, produced a significant reduction of protein synthesis rate in the aorta, and only a modest decrease in mesenteric arteries. Rapamycin, which influences protein synthesis by alternative signaling, had a significant effect in both vessel types. Rapamycin and PD98059 did not alter basal protein synthesis and had minimal effects on arterial pressure.ConclusionERK1/2 and rapamycin-sensitive pathways are involved in pressure-independent angiotensin II-induced vascular protein synthesis in vivo. However, their relative contribution may vary depending on the nature of the artery under investigation.

Endothelin receptor antagonists: novel agents for the treatment of hypertension?

Excitement always greets the development of a new class of therapeutic drug, representing as it does the combined efforts of the pharmaceutical industry, research laboratories and clinicians. Endothelin (ET)-receptor antagonists are being actively developed as new therapeutic agents for cardiovascular diseases, and may also be of use in other pathological conditions. Based on early and indirect evidence, ET has been implicated in the pathophysiology of hypertension; the receptor antagonists have been studied quite extensively in this setting at the preclinical level. We now possess direct evidence that such drugs are effective as antihypertensives in some experimental models of hypertension. Furthermore, the ability of ET-receptor antagonists to prevent hypertension-induced end-organ damage is also well documented. Their capacity to reverse already established target organ alterations remains poorly defined. Based on our current preclinical and clinical knowledge, this review presents the anticipated clinical usefulness of these new drugs, both in terms of blood pressure reduction and the protection of target organs.

An update on the status of endothelin receptor antagonists for hypertension.

Endothelin receptor antagonists (ETRA) are actively developed by the pharmaceutical industry for several cardiovascular indications. In the context of hypertension, preclinical studies are increasingly focused on prevention or regression of end-organ damage and drug combination than on control of arterial pressure in monotherapy, as most experimental models have already been studied. In general, the antihypertensive effect of ETRA is limited but the overwhelming efficacy of this class of drugs to prevent several end-organ damages warrants judicious combination. However, the few studies looking at regression of hypertension-induced cardiovascular alterations proved less successful, suggesting that ETRA should be used early in the treatment of hypertension to obtain full benefit. Judging from the progression of ongoing trials and the development of new trials patients suffering from pulmonary hypertension and heart failure may be the first to benefit from this new class of drugs. However, it is expected that once on the market, responsive subsets of hypertensive patients will be identified and will benefit from end-organ protection.