RESUMO
No disponible
Assuntos
Humanos , Ratos , Animais , Hipertensão/fisiopatologia , Estresse Oxidativo , Espécies Reativas de Oxigênio , Óxido Nítrico/fisiologia , NADPH Oxidases/fisiologia , Ratos Endogâmicos SHR , Aterosclerose/fisiopatologia , Hipertensão/genética , Doenças Cardiovasculares/fisiopatologia , Angiotensina II , Fenótipo , Polimorfismo de Nucleotídeo Único/fisiologiaRESUMO
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No disponible
Assuntos
Humanos , Animais , Ratos , Superóxidos/metabolismo , Doenças Vasculares/metabolismo , Doenças Vasculares/fisiopatologia , Ensaios Clínicos como Assunto , Fatores de RiscoRESUMO
Increased vascular reactive oxygen species production, especially superoxide anion, contributes significantly in the functional and structural alterations present in hypertension. An enhanced superoxide production causes a diminished NO bioavailability by an oxidative reaction that inactivates NO. Exaggerated superoxide levels and a low NO bioavailability lead to endothelial dysfunction and hypertrophy of vascular cells. It has been shown that the enzyme NAD(P)H oxidase plays a major role as the most important source of superoxide anion in vascular cells. Several experimental observations have shown an enhanced superoxide generation as a result of the activation of vascular NAD(P)H oxidase in hypertension. Although this enzyme responds to stimuli such as vasoactive factors, growth factors, and cytokines, some recent data suggest the existence of a genetic background modulating the expression of its different components. New polymorphisms have been identified in the promoter of the p22(phox) gene, an essential subunit of NAD(P)H oxidase, influencing the activity of this enzyme. Genetic investigations of these polymorphisms will provide novel markers for determination of genetic susceptibility to oxidative stress in hypertension.
Assuntos
Hipertensão/fisiopatologia , NADPH Oxidases/metabolismo , Angiotensina II/metabolismo , Animais , Predisposição Genética para Doença , Humanos , Hipertensão Renovascular/fisiopatologia , Músculo Liso Vascular/metabolismo , Estresse Oxidativo , Estresse Mecânico , Regulação para CimaRESUMO
BACKGROUND: Increases in oxidant stress, i.e. excessive production of superoxide anion (O2(.-)), have been reported in different models of hypertension. This study was designed to test the hypothesis that increased O2(.-) production, more than diminished nitric oxide (NO) generation, plays a critical role in endothelial dysfunction present in spontaneously hypertensive rats (SHR). METHODS: The study was performed in 30-week-old normotensive Wistar-Kyoto rats (WKY) and SHR. In addition, 16-week-old SHR were treated with oral irbesartan (average dose 20 mg/kg per day) for 14 weeks (SHR-I). Aortic nicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide phosphate (NADH/NADPH) oxidase activity was determined by use of chemiluminescence with lucigenin. Aortic constitutive nitric oxide synthase (cNOS) activity was determined by measuring the conversion of L-arginine to L-citrulline. Vascular responses to acetylcholine were determined by isometric tension studies. RESULTS: Whereas systolic blood pressure (SBP) was significantly increased in SHR compared with WKY, no differences were observed in SBP between SHR-I and WKY. In SHR compared with WKY, we found significantly greater NADH/NADPH-driven O2(.-) production, similar cNOS-mediated NO production and an impaired vasodilation in response to acetylcholine. Treated SHR had similar NADH/NADPH oxidase activity and significantly lower cNOS activity than the WKY group. Vasodilation in response to acetylcholine was improved in SHR-I. CONCLUSIONS: These findings suggest that a diminished availability of NO secondary to an enhanced NADH/NADPH oxidase-dependent O2(.-) production may play a critical role in endothelial dysfunction of adult SHR.
Assuntos
Endotélio Vascular/fisiopatologia , Hipertensão/fisiopatologia , Músculo Liso Vascular/fisiopatologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico/metabolismo , Superóxidos/metabolismo , Vasodilatação/fisiologia , Acetilcolina/farmacologia , Animais , Anti-Hipertensivos/farmacologia , Aorta/enzimologia , Aorta/fisiologia , Aorta/fisiopatologia , Compostos de Bifenilo/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/fisiologia , Hipertensão/tratamento farmacológico , Hipertensão/metabolismo , Técnicas In Vitro , Irbesartana , Contração Isométrica/efeitos dos fármacos , Contração Isométrica/fisiologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/enzimologia , Músculo Liso Vascular/fisiologia , NADH NADPH Oxirredutases/metabolismo , Óxido Nítrico Sintase Tipo III , Nitroprussiato/farmacologia , Norepinefrina/farmacologia , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Valores de Referência , Tetrazóis/farmacologia , Vasodilatação/efeitos dos fármacosRESUMO
In a previous study, we found that the p22(phox) subunit of the NADH/NADPH oxidase is overexpressed in vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHRs) with enhanced vascular production of superoxide anion ((.)O(2)(-)). Thus, we have investigated whether changes in the sequence or activity of the promoter region of p22(phox) gene are present in SHRs. To carry out this analysis, first of all, we characterized the rat gene structure and promoter region for the p22(phox) subunit. The p22(phox) gene spans approximately 10 kb and contains 6 exons and 5 introns. Primer extension analysis indicated the transcriptional start site 100 bp upstream from the translational start site. The immediate promoter region of the p22(phox) gene does not contain a TATA box, but there are a CCAC box and putative recognition sites for nuclear factors, such as SP1, gamma-interferon, and nuclear factor-kappaB. Using reporter-gene transfection analysis, we found that this promoter was functional in VSMCs. Furthermore, we observed that p22(phox) promoter activity was significantly higher in VSMCs from SHRs than from normotensive Wistar-Kyoto rats. In addition, we found that there were 5 polymorphisms in the sequence of p22(phox) promoter between Wistar-Kyoto rats and SHRs and that they were functional. The results obtained in this study provide a tool to explore the mechanisms that regulate the expression of p22(phox) gene in rat VSMCs. Furthermore, our findings show that changes in the sequence of p22(phox) gene promoter and in the degree of activation of VSMCs are responsible for upregulated expression of p22(phox) in SHRs.
Assuntos
Proteínas de Membrana Transportadoras , Músculo Liso Vascular/metabolismo , NADPH Desidrogenase/genética , NADPH Desidrogenase/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Polimorfismo Genético/genética , Regiões Promotoras Genéticas/genética , Regiões 5' não Traduzidas/genética , Animais , Sequência de Bases , Sítios de Ligação/genética , Células Cultivadas , Clonagem Molecular , Éxons/genética , Genes Reporter , Biblioteca Genômica , Íntrons/genética , Dados de Sequência Molecular , Músculo Liso Vascular/citologia , Mutagênese Sítio-Dirigida , NADPH Oxidases , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Análise de Sequência de DNA , Especificidade da Espécie , Superóxidos/metabolismo , Transfecção , Regulação para Cima/genéticaRESUMO
The term oxidative stress refers to a situation in which cells are exposed to excessive levels of either molecular oxygen or chemical derivatives of oxygen (ie, reactive oxygen species). Three enzyme systems produce reactive oxygen species in the vascular wall: NADH/NADPH oxidase, xanthine oxidoreductase, and endothelial nitric oxide synthase. Among vascular reactive oxygen species superoxide anion plays a critical role in vascular biology because it is the source for many other reactive oxygen species and various vascular cell functions. It is currently thought that increases in oxidant stress, namely excessive production of superoxide anion, are involved in the pathophysiology of endothelial dysfunction that accompanies a number of cardiovascular risk factors including hypercholesterolemia, hypertension and cigarette smoking. On the other hand, vascular oxidant stress plays a pivotal role in the evolution of clinical conditions such as atherosclerosis, diabetes and heart failure.
Assuntos
Vasos Sanguíneos/enzimologia , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Vasos Sanguíneos/metabolismo , Doenças Cardiovasculares/etiologia , Humanos , Complexos Multienzimáticos/metabolismo , NADH NADPH Oxirredutases/metabolismo , Óxido Nítrico/metabolismo , Peróxidos , Superóxidos/metabolismoRESUMO
This study was designed to test the hypothesis that stimulation of nicotinamide adenine dinucleotide/nicotinamide adenine dinucleotide phosphate (NADH/NADPH) oxidase is involved in increased vascular superoxide anion (*O(2)(-)) production in spontaneously hypertensive rats (SHR). The study was performed in 16-week-old and 30-week-old normotensive Wistar-Kyoto rats (WKY(16) and WKY(30), respectively) and in 16-week-old and 30-week-old SHR (SHR(16) and SHR(30), respectively). In addition, 16-week-old SHR were treated with oral irbesartan (average dose 20 mg/kg per day) for 14 weeks (SHR(30)-I). Aortic NADH/NADPH oxidase activity was determined by use of chemiluminescence with lucigenin. The expression of p22phox messenger RNA was assessed by competitive reverse transcription-polymerase chain reaction. Vascular responses to acetylcholine were determined by isometric tension studies. Aortic wall structure was studied, determining the media thickness and the cross-sectional area by morphometric analysis. Whereas systolic blood pressure was significantly increased in the 2 groups of hypertensive animals compared with their normotensive controls, no differences were observed in systolic blood pressure between SHR(30) and SHR(16). No other differences in the parameters measured were found between WKY(16) and SHR(16). In SHR(30) compared with WKY(30), we found significantly greater p22phox mRNA level, NADH/NADPH-driven *O(2)(-) production, media thickness, and cross-sectional area and an impaired vasodilation in response to acetylcholine. Treated SHR had similar NADH/NADPH oxidase activity and p22phox expression as the WKY(30) group. The vascular functional and morphological parameters were improved in SHR(30)-I. These findings suggest that an association exists between p22phox gene overexpression and NADH/NADPH overactivity in the aortas of adult SHR. Enhanced NADH/NADPH oxidase-dependent *O(2)(-) production may contribute to endothelial dysfunction and vascular hypertrophy in this genetic model of hypertension.