Disruption of caveolin-1 leads to enhanced nitrosative stress and severe systolic and diastolic heart failure

doi:10.1016/j.bbrc.2005.12.058

Biochemical and Biophysical Research Communications

Volume 340, Issue 2, 10 February 2006, Pages 702-708

https://doi.org/10.1016/j.bbrc.2005.12.058 Get rights and content

Abstract

Although caveolin-1 is not expressed in cardiomyocytes, this protein is assumed to act as a key regulator in the development of cardiomyopathy. In view of recent discordant findings we aimed to elucidate the cardiac phenotype of independently generated caveolin-1 knockout mice (cav-1^−/−) and to unveil causative mechanisms. Invasive hemodynamic measurements of cav-1^−/− show a severely reduced systolic and diastolic heart function. Additionally, genetic ablation of caveolin-1 leads to a striking biventricular hypertrophy and to a sustained eNOS-hyperactivation yielding increased systemic NO levels. Furthermore, a diminished ATP content and reduced levels of cyclic AMP in hearts of knockout animals were measured. Taken together, these results indicate that genetic disruption of caveolin-1 is sufficient to induce a severe biventricular hypertrophy with signs of systolic and diastolic heart failure. Collectively, our findings suggest a causative role of a sustained nitrosative stress in the development of the pronounced cardiac impairment.

Section snippets

Materials and methods

Animals. Mice were bred and housed in a barrier facility at the Institute for Animal Studies of the University of Technology, Dresden, Germany. They were fed with a standard chow diet and received tap water ad libitum. Unless otherwise indicated mice of two months age were included in the study. Animal experiments conform to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85–23, revised 1996). The generation of the

Cav-1^−/− mice exhibit biventricular hypertrophy

Inspection of cardiac morphology at the age of two months revealed a pronounced biventricular hypertrophy (Fig. 1A). Notably, this hypertrophy develops despite a markedly reduced systemic blood pressure as evidenced by serial cuff-tail measurements (data not shown). To further study the temporal evolution of this distinct cardiac phenotype, we sacrificed mice at the age of two and twelve months and rapidly harvested their hearts. Each heart was washed in PBS, cautiously freed of connective

Discussion

The present study indicates that targeted disruption of caveolin-1 (cav-1) is sufficient to induce a severe cardiomyopathic phenotype with a marked biventricular hypertrophy. By virtue of invasive hemodynamic measurements, we provide compelling evidence for an essential role of cav-1 in the maintenance of both normal systolic and diastolic function in the mammalian heart. Furthermore, we present biochemical evidence for an enhanced eNOS activity yielding increased systemic levels of nitric

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Cited by (59)

Lentinan alleviates diabetic cardiomyopathy by suppressing CAV1/SDHA-regulated mitochondrial dysfunction
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Diabetic cardiomyopathy (DCM), characterized by mitochondrial dysfunction and impaired energetics as contributing factors, significantly contributes to high mortality in patients with diabetes. Targeting key proteins involved in mitochondrial dysfunction might offer new therapeutic possibilities for DCM. Lentinan (LNT), a β-(1,3)-glucan polysaccharide obtained from lentinus edodes, has demonstrated biological activity in modulating metabolic syndrome. In this study, the authors investigate LNT's pharmacological effects on and mechanisms against DCM. The results demonstrate that administering LNT to db/db mice reduces cardiomyocyte apoptosis and mitochondrial dysfunction, thereby preventing DCM. Notably, these effects are fully negated by Caveolin-1 (CAV1) overexpression both in vivo and in vitro. Further studies and bioinformatics analysis uncovered that CAV1 bound with Succinate dehydrogenase subunit A (SDHA), triggering the following ubiquitination and degradation of SDHA, which leads to mitochondrial dysfunction and mitochondria-derived apoptosis under PA condition. Silencing CAV1 leads to reduced apoptosis and improved mitochondrial function, which is blocked by SDHA knockdown. In conclusion, CAV1 directly interacts with SDHA to promote ubiquitination and proteasomal degradation, resulting in mitochondrial dysfunction and mitochondria-derived apoptosis, which was depressed by LNT administration. Therefore, LNT may be a potential pharmacological agent in preventing DCM, and targeting the CAV1/SDHA pathway may be a promising therapeutic approach for DCM.
Caveolin and Endothelial NO Signaling
2018, Current Topics in Membranes
Pulmonary vascular diseases are associated with several factors including infection, cigarette smoking, abuse of dietary suppressants and drugs, prolonged exposure to high altitude, and other causes which in part induce significant oxidative stress resulting in endothelial cell injury, apoptosis, hyperproliferation, and vaso-occlusive disease. Maintenance of normal endothelial cell function is a critical role of endothelial nitric oxide synthase (eNOS) activity and physiologic nitric oxide (NO) signaling in the vascular wall. eNOS expression and activity is regulated by the membrane-associated scaffolding protein caveolin-1 (Cav-1), the main protein constituent of caveolae. This chapter summarizes the literature and highlights unanswered questions related to how inflammation-associated oxidative stress affects Cav-1 expression and regulatory functions, and how dysregulated eNOS enzymatic activity promotes endothelial dysfunction. Focus is given to how the conversion of eNOS from a NO-producing enzyme to a transient oxidant-generating system is associated twith Cav-1 depletion, endothelial cell injury, and pulmonary vascular diseases. Importantly, the vascular defects observed in absence of Cav-1 that give rise to injured or hyperproliferative endothelial cells and promote remodeled vasculature can be rescued by “re-coupling,” inhibiting, or genetically deleting eNOS, supporting the notion that strict control of Cav-1 expression and eNOS activity and signaling is critical for maintaining pulmonary vascular homeostasis.
Endothelial Nitric Oxide Synthase-Induced Hypertrophy and Vascular Dysfunction Contribute to the Left Ventricular Dysfunction in Caveolin-1<sup>−/−</sup> Mice
2017, Canadian Journal of Cardiology
Caveolin-1 (Cav1)^−/− mice display impaired development of left ventricular pressure and increased left ventricular wall thickness but no dilated ventricle; these are typical findings in patients with heart failure with preserved ejection fraction (HfpEF). Aiming to clarify if dysfunctional endothelial nitric oxide synthase (eNOS) influences cardiomyocyte contractility, cardiac conduction system, or afterload/vascular resistance, we studied Cav1^−/−/eNOS^−/− mice.
Cardiac function was assessed in vivo by pressure-volume-catheterization of the left ventricle, echocardiography and electrocardiography. In addition, isolated tissue experiments were performed to evaluate cardiomyocyte contractility (atria) and vessel morphology and function (aorta). Histology, immunoblotting and quantitative polymerase chain reaction were applied to characterise radical formation and oxidative stress in the heart.
Cardiac hypertrophy was completely reversed in Cav1^−/−/eNOS^−/− mice. The impaired pump function in Cav1^−/− mice was significantly improved in Cav1^−/−/eNOS^−/− mice, but no complete alignment with eNOS^−/− controls was achieved, indicating an additional eNOS-independent mechanism contributing to HFpEF in Cav1^−/− mice. It is unlikely that frequently occurring arrhythmias contributed to HFpEF in Cav1^−/− mice. In contrast, numerous eNOS-dependent and eNOS-independent vascular abnomalities could explain the cardiac phenotypes of Cav1^−/− mice.
Synergistic effects between eNOS-related cardiac hypertrophy and vascular hypercontractility appear to underlie the left ventricular dysfunction in Cav1^−/−mice. These findings provide insights relevant to the poorly understood pathophysiology of HFpEF.
Les souris cavéoline-1 (Cav1)^−/− présentent une pression ventriculaire gauche anormale et une augmentation de l’épaisseur de la paroi ventriculaire gauche, sans dilatation du ventricule; ces observations sont caractéristiques des patients atteints d’insuffisance cardiaque à fraction d’éjection préservée (ICFEP). Dans le but de déterminer si un dysfonctionnement de l’oxyde nitrique synthase endothéliale (eNOS) exerce une influence sur la contractilité des cardiomyocytes, le système de conduction cardiaque ou la résistance vasculaire/postcharge, nous avons étudié les souris Cav1^−/−/eNOS^−/−.
La fonction cardiaque a été évaluée in vivo par des mesures de la pression et des volumes via cathétérisme du ventricule gauche, échocardiographie et électrocardiographie. De plus, des expériences sur des tissus isolés ont été réalisées pour évaluer la contractilité des cardiomyocytes (oreillette) ainsi que la morphologie et la fonction des vaisseaux (aorte). L'histologie, l'immunobuvardage et la réaction en chaîne par polymérase en temps réel ont été effectués pour discerner la formation de radicaux et le stress oxydatif dans le cœur.
L’hypertrophie cardiaque a été complètement inversée chez les souris Cav1^−/−/eNOS. La fonction de pompage, altérée chez les souris Cav1^−/−, a été significativement améliorée dans le groupe Cav1^−/−/eNOS^−/−, mais l’alignement parfait sur les témoins eNOS^−/− n’a pas été obtenu, ce qui laisse croire à l’existence d’un mécanisme additionnel indépendant de l’enzyme eNOS qui contribuerait à l’ICFEP chez les souris Cav1^−/−. Il est peu probable que les arythmies fréquentes aient contribué à l’ICFEP chez les souris Cav1^−/−. En revanche, plusieurs anomalies vasculaires dépendantes et indépendantes de l’eNOS pourraient expliquer les phénotypes cardiaques des souris Cav1^−/−.
Des effets synergiques entre l’hypertrophie cardiaque et l’hypercontractilité vasculaire liées à l’eNOS semblent être à l’origine de la dysfonction ventriculaire gauche chez les souris Cav1^−/−. Ces résultats fournissent certains éclaircissements au sujet de la physiopathologie de l’ICFEP, qui reste mal connue.
Combinatorial therapy with acetylation and methylation modifiers attenuates lung vascular hyperpermeability in endotoxemia-induced mouse inflammatory lung injury
2014, American Journal of Pathology
Impairment of tissue fluid homeostasis and migration of inflammatory cells across the vascular endothelial barrier are crucial factors in the pathogenesis of acute lung injury (ALI). The goal for treatment of ALI is to target pathways that lead to profound dysregulation of the lung endothelial barrier. Although studies have shown that chemical epigenetic modifiers can limit lung inflammation in experimental ALI models, studies to date have not examined efficacy of a combination of DNA methyl transferase inhibitor 5-Aza 2-deoxycytidine and histone deacetylase inhibitor trichostatin A (herein referred to as Aza+TSA) after endotoxemia-induced mouse lung injury. We tested the hypothesis that treatment with Aza+TSA after lipopolysaccharide induction of ALI through epigenetic modification of lung endothelial cells prevents inflammatory lung injury. Combinatorial treatment with Aza+TSA mitigated the increased endothelial permeability response after lipopolysaccharide challenge. In addition, we observed reduced lung inflammation and lung injury. Aza+TSA also significantly reduced mortality in the ALI model. The protection was ascribed to inhibition of the eNOS-Cav1-MLC2 signaling pathway and enhanced acetylation of histone markers on the vascular endothelial-cadherin promoter. In summary, these data show for the first time the efficacy of combinatorial Aza+TSA therapy in preventing ALI in lipopolysaccharide-induced endotoxemia and raise the possibility of an essential role of DNA methyl transferase and histone deacetylase in the mechanism of ALI.
Altered angiogenesis in caveolin-1 gene-deficient mice is restored by ablation of endothelial nitric oxide synthase
2012, American Journal of Pathology
Caveolin-1 is an essential structural protein of caveolae, specialized plasma membrane organelles highly abundant in endothelial cells, where they regulate multiple functions including angiogenesis. Caveolin-1 exerts a tonic inhibition of endothelial nitric oxide synthase (eNOS) activity. Accordingly, caveolin-1 gene–disrupted mice have enhanced eNOS activity as well as increased systemic nitric oxide (NO) levels. We hypothesized that excess eNOS activity, secondary to caveolin deficiency, would mediate the decreased angiogenesis observed in caveolin-1 gene–disrupted mice. We tested tumor angiogenesis in mice lacking either one or both proteins, using in vitro, ex vivo, and in vivo assays. We show that endothelial cell migration, tube formation, cell sprouting from aortic rings, tumor growth, and angiogenesis are all significantly impaired in both caveolin-1–null and eNOS-null mice. We further show that these parameters were either partially or fully restored in double knockout mice that lack both caveolin-1 and eNOS. Furthermore, the effects of genetic ablation of eNOS are mimicked by the administration of the NOS inhibitor N-nitro-l-arginine methyl ester hydrochloride (L-NAME), including the reversal of the caveolin-1–null mouse angiogenic phenotype. This study is the first to demonstrate the detrimental effects of unregulated eNOS activity on angiogenesis, and shows that impaired tumor angiogenesis in caveolin-1–null mice is, at least in part, the result of enhanced eNOS activity.
Study on Potential Differentially Expressed Genes in Idiopathic Pulmonary Fibrosis by Bioinformatics and Next-Generation Sequencing Data Analysis
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^☆: Abbreviations: eNOS, endothelial nitric oxide synthase; cAMP, cyclic adenosine monophosphate; wt, wild-type; cav-1^−/−, caveolin-1 knockout animals; ATP, adenosine triphosphate; ANP, atrial natriuretic peptide.

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Disruption of caveolin-1 leads to enhanced nitrosative stress and severe systolic and diastolic heart failure☆

Abstract

Section snippets

Materials and methods

Cav-1−/− mice exhibit biventricular hypertrophy

Discussion

Cell

J. Biol. Chem.

J. Mol. Cell Cardiol.

FEBS Lett.

J. Biol. Chem.

The fine structure of the gall bladder epithelium of the mouse

J. Biophys. Biochem. Cytol.

Loss of caveolae, vascular dysfunction, and pulmonary defects in caveolin-1 gene-disrupted mice

Science

Caveolin-deficient mice: insights into caveolar function human disease

J. Clin. Invest.

In vivo delivery of the caveolin-1 scaffolding domain inhibits nitric oxide synthesis and reduces inflammation

Nat. Med.

Caveolae and caveolins in the cardiovascular system

Circ. Res.

Caveolin-1 null mice develop cardiac hypertrophy with hyperactivation of p42/44 MAP kinase in cardiac fibroblasts

Am. J. Physiol. Cell Physiol.

Cav-1^−/− mice exhibit biventricular hypertrophy