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1.
Int J Urol ; 28(1): 115-124, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33289131

RESUMO

OBJECTIVE: To assess the characteristics of adipose-derived regenerative cells, and provide supportive data explaining the mechanism of efficacy observed for the use of these cells in the treatment of stress urinary incontinence. METHODS: Adipose tissues were harvested by abdominal liposuction from healthy donors and patients with stress urinary incontinence. Adipose-derived regenerative cells were isolated from tissues using the Celution system, and assessed for their characteristics and ability to differentiate into smooth muscle cells. RESULTS: Adipose-derived regenerative cells isolated by the Celution system developed into fibroblastic colonies. Flow cytometric analysis of adipose-derived stem cell markers showed that adipose-derived regenerative cells were positive for CD34 and CD44, and negative for CD31. Immunofluorescence staining after differentiation showed that colony-forming cells were positive for alpha-smooth muscle actin, calponin and desmin, which are smooth muscle cell markers. A cytokine release assay showed that adherent cells secreted cytokines associated with angiogenesis, including vascular endothelial growth factor-A, angiopoietin-2 and placental growth factor. CONCLUSIONS: Adipose-derived regenerative cells collected by the Celution system might have clonogenic capacity and an angiogenetic function. These properties might contribute to the mechanisms through which regenerative cell therapy by periurethral injection of autologous adipose-derived regenerative cells ameliorates stress urinary incontinence.


Assuntos
Incontinência Urinária por Estresse , Tecido Adiposo , Células Cultivadas , Feminino , Humanos , Masculino , Fator de Crescimento Placentário , Uretra/cirurgia , Incontinência Urinária por Estresse/terapia , Fator A de Crescimento do Endotélio Vascular
2.
J Pharmacol Exp Ther ; 337(3): 591-9, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21357660

RESUMO

The elderly are prone to postprandial hyperglycemia that increases their cardiovascular risk. Although insulin therapy is necessary to treat diabetes, high plasma concentrations of insulin may cause the development of atherosclerosis and accelerate endothelial senescence. We assumed that high glucose causes stress-induced premature senescence and replicative senescence and examined the regulatory role of insulin in endothelial senescence and functions under different glucose conditions. Exposure of human endothelial cells to high glucose (22 mM) for 3 days increased senescence-associated-ß-galactosidase activity, a senescence marker, and decreased telomerase activity, a replicative senescence marker. Physiological concentrations of insulin preserved telomere length and delayed endothelial senescence under high-glucose conditions. The effect of insulin under high-glucose conditions was associated with reduced reactive oxygen species and increased nitric oxide (NO). Small interfering RNA targeting endothelial NO synthase reduced the antisenescence effects of insulin. Physiological concentrations of insulin also reversed high glucose-induced increases in p53 and vascular cell adhesion molecule-1 and decreases in senescence marker protein-30. On the other hand, when insulin was given at any concentrations under normal glucose or at high concentrations under high glucose, its ability to promote cellular senescence was unrelated to endothelial NO. Finally, streptozotocin-induced diabetes showed more senescent cells in the aortic endothelium of aged rats compared with age-matched control and insulin-treated animals. Conclusively, the regulatory effects of insulin on endothelial senescence were modulated by the glucose environment. These data may help explain insulin's complicated roles in atherosclerosis in the elderly.


Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Insulina/farmacologia , Óxido Nítrico/metabolismo , Telômero/fisiologia , Envelhecimento/metabolismo , Animais , Aorta , Aterosclerose/fisiopatologia , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Glucose , Humanos , Hipoglicemiantes/metabolismo , Insulina/fisiologia , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , RNA Interferente Pequeno/genética , Ratos , Telomerase/metabolismo , Veias Umbilicais/efeitos dos fármacos , Veias Umbilicais/metabolismo , beta-Galactosidase/metabolismo
3.
Life Sci ; 85(7-8): 316-21, 2009 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-19560472

RESUMO

AIMS: This study evaluated the presence of genetic mutations in relation to thrombosis or atherosclerosis in elderly women. MAIN METHODS: This is an observational study of 93 Japanese women with a mean age of 80.9 years recruited from outpatient clinics of Nagoya University and its related hospitals. Ten single nucleotide polymorphisms (SNPs) were studied. Each gene studied acts in or is related to either blood coagulation (factor V Leiden, prothrombin G20210A, factor XIII Val34Leu, factor VII Arg353Gln, MTHFR C677T, beta-fibrinogen G-455A, PAI-1 4G/5G), metabolic syndrome-related pathways (PPARalpha Leu162Val), or endothelium/estrogen system (eNOS Glu298Asp, ERalpha IVS1-401). SNPs were analyzed for their relation to clinical values including lipids, B-type natriuretic peptide (BNP), fasting plasma glucose, tumor necrosis factor-alpha, interleukin-6, cyclic GMP, and nitric oxide metabolites. KEY FINDINGS: Comparisons between the distributions of different genotypes and clinical values showed three relationships. First, factor VII Arg353Gln and HDL-cholesterol (HDL-C) were linked to Arg/Arg carriers at higher levels (P=.049). The HDL-C to LDL-cholesterol ratio supported this link (P=.027). Second, eNOS Glu298Asp and triglycerides were linked to Glu/Glu carriers at higher levels (P=.031). Third, ERalpha IVS1-401 and BNP were related to CC genotype at lower levels (P=.031). Additionally, the last two relations showed that genotype does not influence the demarcation line of biomarkers, but the plasma/serum levels of biomarkers instead. SIGNIFICANCE: Correlations of factor VII Arg353Gln with HDL-C and eNOS Glu298Asp with triglycerides are new findings. Polymorphisms in the endothelium/estrogen system and the heart failure marker BNP are also correlated, with ERalpha IVS1-401 being the first identified marker. SNPs may be helpful for understanding the pathophysiology of atherosclerotic diseases in elderly women.


Assuntos
Envelhecimento/genética , Aterosclerose/genética , Peptídeo Natriurético Encefálico/sangue , Polimorfismo de Nucleotídeo Único , Trombose/genética , Idoso , Idoso de 80 Anos ou mais , Aterosclerose/sangue , DNA/genética , Receptor alfa de Estrogênio/genética , Fator VII/genética , Feminino , Genótipo , Humanos , Japão , Pessoa de Meia-Idade , Óxido Nítrico Sintase Tipo III/genética , Trombose/sangue
4.
Pharmacol Ther ; 120(3): 333-9, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18930078

RESUMO

Cellular senescence is characterized by permanent exit from the cell cycle and the appearance of distinct morphological and functional changes associated with an impairment of cellular homeostasis. Many studies support the occurrence of vascular endothelial cell senescence in vivo, and the senescent phenotype of endothelial cells can be transformed from anti-atherosclerotic to pro-atherosclerotic. Thus, endothelial cell senescence promotes endothelial dysfunction and may contribute to the pathogenesis of age-associated vascular disorders. Emerging evidence suggests that increasing nitric oxide (NO) bioavailability or endothelial NO synthase (eNOS) activity activates telomerase and delays endothelial cell senescence. In this review, we discuss the potential mechanisms underlying the ability of NO to prevent endothelial cell senescence and describe the possible changes in the NO-mediated anti-senescence effect under pathophysiological conditions, including oxidative stress and hyperglycemia. Further understanding of the mechanisms underlying the anti-senescence effect of NO in endothelial cells will provide insights into the potential of eNOS-based anti-senescence therapy for age-associated vascular disorders.


Assuntos
Senescência Celular/fisiologia , Endotélio Vascular/fisiologia , Óxido Nítrico/fisiologia , Animais , Endotélio Vascular/metabolismo , Estrogênios/fisiologia , Glucose/metabolismo , Humanos , Telômero/fisiologia
5.
Am J Physiol Heart Circ Physiol ; 293(1): H790-7, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17449545

RESUMO

The long-term benefits of nitroglycerin therapy are limited by tolerance development. Understanding the precise nature of mechanisms underlying nitroglycerin-induced endothelial cell dysfunction may provide new strategies to prevent tolerance development. In this line, we tested interventions to prevent endothelial dysfunction in the setting of nitrate tolerance. When bovine aortic endothelial cells (BAECs) were continuously treated with nitric oxide (NO) donors, including nitroglycerin, over 2-3 days, basal production of nitrite and nitrate (NO(x)) was diminished. The diminished basal NO(x) levels were mitigated by intermittent treatment allowing an 8-h daily nitrate-free interval during the 2- to 3-day treatment period. Addition of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor apocynin restored the basal levels of NO(x) that were decreased by continuous nitroglycerin treatment of BAECs. Apocynin caused significant improvement of increased mRNA and protein levels of endothelial nitric oxide synthase (eNOS) in BAECs given nitroglycerin continuously over the treatment period. Apocynin also reduced endothelial production of reactive oxygen species (ROS) after continuous nitroglycerin treatment. These results showed an essential similarity to the effects of a nitrate-free interval. Application of the NOS inhibitor N(omega)-nitro- l-arginine methyl ester caused a recovery effect on basal NO(x) and eNOS expression but was without effect on ROS levels in continuously NO donor-treated BAECs. In conclusion, the present study characterized abnormal features and functions of endothelial cells following continuous NO donor application. We suggest that inhibition of NADPH oxidase, by preventing NO donor-induced endothelial dysfunction, may represent a potential therapeutic strategy that confers protection from nitrate tolerance development.


Assuntos
Acetofenonas/administração & dosagem , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , NADPH Oxidases/antagonistas & inibidores , Nitratos/metabolismo , Doadores de Óxido Nítrico/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Animais , Bovinos , Células Cultivadas , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Tolerância a Medicamentos/fisiologia , Inibidores Enzimáticos/administração & dosagem
6.
Life Sci ; 80(6): 592-9, 2007 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-17141277

RESUMO

UNLABELLED: The effect of beta antagonists in the diabetic vascular lesion is controversial. We investigated the effect of celiprolol hydrochloride, a beta1 antagonist and mild beta2 agonist, on the lesions and function in type II male Otsuka Long-Evans Tokushima Fatty (OLETF) diabetic rats. OLETF rats were fed regular chow with or without atenolol (25 mg/kg/day) or celiprolol (100 mg/kg/day) treatment (group DM, no treatment; group DM-a, atenolol treatment; group DM-c, celiprolol treatment), and treatment was continued for 31 days. Separately, normoglycemic control rats, LETO, were prepared as group C. On day 3, endothelial cells of the right internal carotid artery were removed by balloon injury, and the rats were evaluated 4 weeks after balloon injury. The plasma glucose and lipid levels were unchanged throughout the treatment period. Intimal thickening was observed in the right carotid artery in the DM and DM-a groups; however, little thickening was observed in those of DM-c rats. Acetylcholine-induced NO-dependent relaxation in arteries was improved in DM-c rats compared with DM and DM-a rats (maximum relaxation DM 30.8+/-4.5, DM-a 37.4+/-3.9, DM-c 48.8+/-4.6%, *P<0.05 vs. DM for DM-c rats). Tone-related basal NO release and acetylcholine-induced NO-dependent relaxation in the arteries and plasma NO(x) (sum of NO(2)(-) and NO(3)(-)) were greater in DM-c and C groups than in DM and DM-a groups. The serum TNFalpha levels did not increase in DM-c rats compared with those of the DM or DM-a groups, and were comparable with those of group C. CONCLUSION: In conclusion, Celiprolol improves endothelial function in the arteries of OLETF rats, and further restore it 4 weeks after endothelial denudation in the arteries of OLETF rats. NO and O(2)(-) may have a role in the important underlying mechanisms by reducing the TNFalpha levels.


Assuntos
Antagonistas de Receptores Adrenérgicos beta 1 , Agonistas de Receptores Adrenérgicos beta 2 , Celiprolol/uso terapêutico , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Tipo 2/complicações , Endotélio Vascular/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Aorta Abdominal/metabolismo , Aterosclerose/metabolismo , Aterosclerose/prevenção & controle , Pressão Sanguínea/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Celiprolol/farmacologia , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Angiopatias Diabéticas/metabolismo , Angiopatias Diabéticas/prevenção & controle , Masculino , Nitratos/sangue , Óxido Nítrico/metabolismo , Nitritos/sangue , Ratos , Ratos Long-Evans , Superóxidos/metabolismo
7.
Proc Natl Acad Sci U S A ; 103(45): 17018-23, 2006 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-17075048

RESUMO

Senescence may contribute to the pathogenesis of atherosclerosis. Although the bioavailability of nitric oxide (NO) is limited in senescence, the effect of NO on senescence and its relationship to cardiovascular risk factors have not been investigated fully. We studied these factors by investigating senescence-associated beta-galactosidase (SA-beta-gal) and human telomerase activity in human umbilical venous endothelial cells (HUVECs). Treatment with NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-aminoethyl)amino]diazen-1-ium-1,2-diolate (DETA-NO) and transfection with endothelial NO synthase (eNOS) into HUVECs each decreased the number of SA-beta-gal positive cells and increased telomerase activity. The NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) abolished the effect of eNOS transfection. The physiological concentration of 17beta-estradiol activated hTERT, decreased SA-beta-gal-positive cells, and caused cell proliferation. However, ICI 182780, an estrogen receptor-specific antagonist, and L-NAME each inhibited these effects. Finally, we investigated the effect of NO bioavailability on high glucose-promoted cellular senescence of HUVECs. Inhibition by eNOS transfection of this cellular senescence under high glucose conditions was less pronounced. Treatment with L-arginine or L-citrulline of eNOS-transfected cells partially inhibited, and combination of L-arginine and L-citrulline with antioxidants strongly prevented, high glucose-induced cellular senescence. These data demonstrate that NO can prevent endothelial senescence, thereby contributing to the anti-senile action of estrogen. The ingestion of NO-boosting substances, including L-arginine, L-citrulline, and antioxidants, can delay endothelial senescence under high glucose. We suggest that the delay in endothelial senescence through NO and/or eNOS activation may have clinical utility in the treatment of atherosclerosis in the elderly.


Assuntos
Senescência Celular/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Óxido Nítrico/farmacologia , Aterosclerose/etiologia , Aterosclerose/metabolismo , Aterosclerose/patologia , Aterosclerose/prevenção & controle , Células Cultivadas , Angiopatias Diabéticas/etiologia , Angiopatias Diabéticas/metabolismo , Angiopatias Diabéticas/patologia , Angiopatias Diabéticas/prevenção & controle , Células Endoteliais/metabolismo , Feminino , Humanos , Menopausa , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Espécies Reativas de Oxigênio/metabolismo , Telomerase/metabolismo , Transfecção
8.
Atherosclerosis ; 187(2): 316-24, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16325187

RESUMO

OBJECTIVE: We have reported that inducible nitric oxide synthase (iNOS) is present only in deep areas of plaque in atherosclerosis. However, the role of iNOS in the development of atherosclerosis is not well known. We therefore investigated the relevance of iNOS inhibition. METHODS AND RESULTS: Seven groups of male rabbits were fed a 0.5% high-cholesterol diet (HCD) for 8 weeks. Gp1-HCD was fed HCD only; Gp2-O17 was fed HCD with ONO1714, an iNOS inhibitor; Gp3-AG was fed HCD with amino-guanidine (AG), an iNOS inhibitor; Gp4-AR was fed HCD with l-arginine; Gp5-AR-O17 was fed HCD with l-arginine with ONO1714; Gp6-LNA was fed HCD with l-NAME (a NOS inhibitor); and Gp7-LN-O17 was fed HCD with l-NAME plus ONO1714. ONO1714 decreased atherosclerosis by about 70% (area occupied by lesions: 3.0+/-0.4% in Gp2-O17 versus 10.3+/-1.6% in Gp1-HCD) and also decreased atherosclerosis in Gp7-LN-O17. The ONO compound enhanced the atheroprotective effect of l-arginine. Amino-guanidine also showed an anti-atherosclerotic effect. Tone-related basal NO release and acetylcholine-induced NO-dependent relaxation were improved in Gp2-O17 and Gp5-AR-O17. O(2)(-) release was decreased in Gp2-O17 and Gp7-LN-O17. CONCLUSION: ONO1714 retards the progression of atherosclerosis in rabbits. Although the up-regulation of endothelial nitric oxide synthase (eNOS) and the decrease of O(2)(-) may play roles in this retardation, the inhibition of iNOS may be the principal factor, alone was not sufficient.


Assuntos
Amidinas/farmacologia , Aterosclerose/prevenção & controle , Colesterol na Dieta/farmacologia , Inibidores Enzimáticos/farmacologia , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Animais , Aorta Torácica/enzimologia , Aorta Torácica/patologia , Arginina/farmacologia , Aterosclerose/tratamento farmacológico , Aterosclerose/patologia , Western Blotting , Colesterol na Dieta/sangue , GMP Cíclico/metabolismo , Endotélio Vascular/enzimologia , Endotélio Vascular/patologia , Compostos Heterocíclicos com 2 Anéis/farmacologia , Imuno-Histoquímica , Macrófagos/patologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , RNA Mensageiro/metabolismo , Coelhos , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos
9.
Proc Natl Acad Sci U S A ; 102(38): 13681-6, 2005 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-16157883

RESUMO

The objective of this study was to evaluate the influence of ingested l-arginine, l-citrulline, and antioxidants (vitamins C and E) on the progression of atherosclerosis in rabbits fed a high-cholesterol diet. The fatty diet caused a marked impairment of endothelium-dependent vasorelaxation in isolated thoracic aorta and blood flow in rabbit ear artery in vivo, the development of atheromatous lesions and increased superoxide anion production in thoracic aorta, and increased oxidation-sensitive gene expression [Elk-1 and phosphorylated cAMP response element-binding protein]. Rabbits were treated orally for 12 weeks with l-arginine, l-citrulline, and/or antioxidants. l-arginine plus l-citrulline, either alone or in combination with antioxidants, caused a marked improvement in endothelium-dependent vasorelaxation and blood flow, dramatic regression in atheromatous lesions, and decrease in superoxide production and oxidation-sensitive gene expression. These therapeutic effects were associated with concomitant increases in aortic endothelial NO synthase expression and plasma NO(2)(-)+NO(3)(-) and cGMP levels. These observations indicate that ingestion of certain NO-boosting substances, including l-arginine, l-citrulline, and antioxidants, can abrogate the state of oxidative stress and reverse the progression of atherosclerosis. This approach may have clinical utility in the treatment of atherosclerosis in humans.


Assuntos
Arginina/administração & dosagem , Arteriosclerose/metabolismo , Citrulina/administração & dosagem , Dieta Aterogênica , Endotélio Vascular/metabolismo , Animais , Antioxidantes/administração & dosagem , Aorta Torácica/metabolismo , Aorta Torácica/patologia , Arteriosclerose/tratamento farmacológico , Arteriosclerose/patologia , Ácido Ascórbico/administração & dosagem , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/biossíntese , GMP Cíclico/metabolismo , Proteínas de Ligação a DNA/biossíntese , Endotélio Vascular/patologia , Humanos , Masculino , Nitratos/metabolismo , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo III , Nitritos/metabolismo , Oxirredução/efeitos dos fármacos , Proteínas Proto-Oncogênicas/biossíntese , Coelhos , Superóxidos/metabolismo , Fatores de Transcrição/biossíntese , Vasodilatação/efeitos dos fármacos , Vitamina E/administração & dosagem , Proteínas Elk-1 do Domínio ets
10.
Atherosclerosis ; 176(2): 255-63, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15380447

RESUMO

BACKGROUND: The remarkable anti-atherosclerotic effects of 3-hydroxy-3-methyl-glutaryl-CoA reductase inhibitor have not been demonstrated in diet induced severe hyperlipidemia in rabbit model. OBJECTIVE: We have investigated the effect of pitavastatin, a newly developed statin, on atherosclerosis in rabbits. METHODS AND RESULTS: Oophorectomized female NZW rabbits were fed 0.3% cholesterol chow for 12 weeks with or without pitavastatin (0.1mg/kg per day) (Gp.NK and HCD). The level of serum cholesterol was decreased in Gp.NK compared with Gp.HCD (772.8 +/- 70.2 versus 1056.9 +/- 108.3 mg/d), whereas no significant alterations were observed in triglyceride and HDL-cholesterol. NO dependent response stimulated by acetylcholine and calcium ionophore A23187 and tone related basal NO response induced by N(G)-monomethyl-l-arginine acetate were all improved by pitavastatin treatment. Pitavastatin treatment increased the level of cyclic GMP in the aorta of cholesterol fed rabbits. In the aorta, the expression of eNOS mRNA was significantly up regulated and O(2)(-) production was slightly reduced in Gp.NK animals. Atherosclerotic area was significantly decreased in aortic arch and thoracic aorta from Gp.NK compared with those from Gp.HCD ( 15.1 +/- 5.3 versus 41.9 +/- 10.2%, 3.1 +/- 1.1versus 7.9 +/- 1.2% in Gp.NK and Gp.HCD aortic arch and thoracic aorta). Anti-macrophage staining area, the MMP1 or 2 and the nitrotyrosine positive area were decreased in Gp.NK. CONCLUSION: Pitavastatin retards the progression of atherosclerosis formation and it improves NO bioavailability by eNOS up-regulation and decrease of O(2)(-).


Assuntos
Arteriosclerose/tratamento farmacológico , Arteriosclerose/fisiopatologia , Inibidores Enzimáticos/farmacologia , Quinolinas/farmacologia , Administração Oral , Animais , Arteriosclerose/veterinária , Disponibilidade Biológica , HDL-Colesterol/sangue , Modelos Animais de Doenças , Progressão da Doença , Feminino , Óxido Nítrico/farmacologia , Coelhos , Triglicerídeos/sangue , Regulação para Cima
11.
Biochim Biophys Acta ; 1693(3): 185-91, 2004 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-15363632

RESUMO

Human inducible nitric oxide synthase (iNOS) is most readily observed in macrophages from patients with inflammatory diseases like atherosclerosis. The aim of the present study was to find out the combined effect of male sex hormone; testosterone and apocynin (NADPH oxidase inhibitor) on cytokine-induced iNOS production. THP-1 cells were differentiated into macrophages by phorbol myristate acetate (PMA). Expression of iNOS was induced by the addition of cytokine mixture? Testosterone was added at different concentrations (10(-6)-10(-12) M) with apocynin (1 mM). Testosterone (10(-8), 10(-10) M) inhibited NOx production in cytokine-added THP-1 cells which was further confirmed by quantikine assay of iNOS protein and RT-PCR analysis. Testosterone treatment decreased 40% of superoxide anion production. Testosterone showed inhibition of NADPH oxidase, especially expression of p67phox and p47phox (cytosol subunits). Addition of testosterone with apocynin further decreased the expression of p67phox and p47phox subunits of NADPH oxidase. The findings of the present study suggest that, testosterone; the male androgen plays an important role in the prevention of atherogenesis. Even though apocynin does not have any role on NO production, addition of apocynin together with testosterone is effective in suppressing iNOS activity.


Assuntos
Acetofenonas/farmacologia , Macrófagos/metabolismo , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico/metabolismo , Superóxidos/metabolismo , Testosterona/farmacologia , Acetofenonas/administração & dosagem , Inibidores da Aromatase , Células Cultivadas , Inibidores Enzimáticos/farmacologia , Fadrozol/farmacologia , Humanos , Macrófagos/efeitos dos fármacos , Masculino , NADPH Oxidases/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II , Fosfoproteínas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Testosterona/administração & dosagem , Acetato de Tetradecanoilforbol
12.
Cardiovasc Res ; 61(2): 339-51, 2004 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-14736551

RESUMO

The effects of in vivo gene transfer of endothelial nitric oxide synthase (eNOS) and inducible NOS (iNOS) genes on severe atherosclerosis were investigated in rabbits. The recombinant adenoviruses, Ad.eNOS and Ad.iNOS, which respectively express eNOS and iNOS, were constructed. Atherosclerosis was induced by a balloon injury followed by a high cholesterol diet for 12 weeks. The rabbits were divided into six groups: Gp cont (no treatment); Gp null (adenovirus sham-infected); Gp eNOS (Ad.eNOS); Gp iNOS (Ad.iNOS); Gp e+i (Ad.eNOS plus Ad.iNOS); and Gp heNOS (a high dose of Ad.eNOS). Examinations were carried out 7 days after gene transfer. Plasma lipid levels were not significantly changed, but transfection with Ad.eNOS (Gp eNOS and Gp heNOS) decreased the tissue cholesterol concentration and regressed atherosclerotic lesions. Vessels treated with Ad.iNOS (Gp iNOS and Gp e+i) showed iNOS staining in the atheroma, and slight staining at other parts of the vessels; those treated with Ad.eNOS showed eNOS staining in the endothelium and subintima, and slight staining at other parts. Ad.eNOS transfection, but not Ad.iNOS or Ad.eNOS+Ad.iNOS transfection, improved the impaired aortic endothelium-dependent relaxation (EDR) and basal NO-dependent response, increased tissue cyclic GMP (cGMP), and decreased the release of O2- from vessels. eNOS treatment showed a decreasing tendency in regions with peroxynitrite staining, MMP1 staining, and suspected apoptosis. In conclusion, in vivo gene transfer of eNOS, but not iNOS or eNOS plus iNOS, regressed atherosclerosis. The relations among NO, O2-, and peroxynitrite may be critical, and lipid resorption from the lesions may be responsible for the regression.


Assuntos
Arteriosclerose/terapia , Terapia Genética/métodos , Óxido Nítrico Sintase/genética , Transdução Genética/métodos , Acetilcolina , Adenoviridae/genética , Animais , Aorta Abdominal/metabolismo , Aorta Abdominal/patologia , Arteriosclerose/metabolismo , Arteriosclerose/patologia , Colesterol/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Vetores Genéticos/administração & dosagem , Infusões Intra-Arteriais , Metabolismo dos Lipídeos , Masculino , Modelos Animais , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , Óxido Nítrico Sintase Tipo III , Nitroglicerina , Oxigênio/metabolismo , Ácido Peroxinitroso/metabolismo , Coelhos , Vasodilatadores , ômega-N-Metilarginina/farmacologia
13.
Biochim Biophys Acta ; 1640(2-3): 113-8, 2003 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-12729920

RESUMO

In this report, we demonstrate that NADPH oxidase is activated by tumor necrosis factor-alpha (TNF-alpha) plus interferon-gamma (IFN-gamma) in human monocytic cells (THP-1 cells) differentiated with phorbol ester (PMA) and that physiological concentration of 17beta-estradiol inhibits NADPH oxidase activity in THP-1 cells stimulated with TNF-alpha plus IFN-gamma. This effect is mediated by estrogen receptor based on estrogen receptor antagonist (ICI 182, 780) that diminishes inhibition by 17beta-estradiol. This inhibition is specific in 17beta-estradiol because 17alpha-estradiol, testosterone and progesterone do not inhibit NADPH oxidase activity. Activation of NADPH oxidase induced by TNF-alpha plus IFN-gamma is caused by up-regulation of p47(phox) (cytosolic component of NADPH oxidase) expression. 17beta-Estradiol prevents the up-regulation of p47(phox) mRNA and protein expression. This prevention of p47(phox) expression depends on the inhibition of NF-kappaB activation. Our results implicate that 17beta-estradiol has an anti-atherosclerotic effects through the improvement of nitric oxide (NO) bioavailability caused by the regulation of superoxide (O(2)(-)) production.


Assuntos
Estradiol/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , NADPH Oxidases/antagonistas & inibidores , Fosfoproteínas/genética , Linhagem Celular , Humanos , Interferon gama/farmacologia , NF-kappa B/antagonistas & inibidores , RNA Mensageiro/análise , Fator de Necrose Tumoral alfa/farmacologia
14.
Atherosclerosis ; 168(1): 23-31, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12732383

RESUMO

Although sarpogrelate HCl is widely used for the prevention of arterial thrombosis, its effect on atherosclerosis is unknown. Accordingly, we here investigated the effects of sarpogrelate HCl on a rabbit model of atherosclerosis. Male rabbits were fed a 0.5% cholesterol diet (HCD) (Gp 1), HCD with vitamin E (Gp 2), HCD with vitamin E and sarpogrelate (Gp 3), or HCD with sarpogrelate alone (Gp 4) for 8 weeks. The atherosclerotic area was decreased by feeding of vitamin E and sarpogrelate (16.9+/-2.0% in Gp 1 vs. 8.2+/-2.0% in Gp 3). Tone-related basal NO release was higher in Gps 3 and 4. Acetylcholine-induced relaxation tended to be improved in Gp 3. The amount of eNOS mRNA was increased in Gp 4, and aortic cyclic GMP concentration showed the same tendency. O(2)(-) release tended to be decreased in Gps 2 and 3. The matrix metalloproteinase-1 (MMP-1)-positive area was decreased, and the percentage ratio of cell numbers of smooth muscle cells/macrophages in the plaque was increased in Gp 3. The results demonstrated that sarpogrelate HCl retards the progression of atherosclerosis in rabbits, and that this effect is enhanced by concomitant administration of vitamin E. Although upregulation of eNOS may play a role as one of the underlying mechanisms, our results suggest that an additional mechanism-possibly involving the antiproliferative effects of sarpogrelate HCl on smooth muscle cells and macrophages-may also play an important role.


Assuntos
Doença da Artéria Coronariana/tratamento farmacológico , Antagonistas da Serotonina/uso terapêutico , Succinatos/uso terapêutico , Acetilcolina/administração & dosagem , Animais , Antioxidantes/uso terapêutico , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/patologia , Aorta Torácica/fisiopatologia , Biomarcadores/sangue , Proteínas Sanguíneas/metabolismo , HDL-Colesterol/sangue , Doença da Artéria Coronariana/etiologia , Doença da Artéria Coronariana/metabolismo , GMP Cíclico/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Relação Dose-Resposta a Droga , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/patologia , Endotélio Vascular/fisiopatologia , Hipercolesterolemia/tratamento farmacológico , Hipercolesterolemia/etiologia , Hipercolesterolemia/metabolismo , Masculino , Metaloproteinase 1 da Matriz/efeitos dos fármacos , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Metaloproteinase 2 da Matriz/metabolismo , Modelos Cardiovasculares , Contração Miocárdica/efeitos dos fármacos , Contração Miocárdica/fisiologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Óxido Nítrico Sintase/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II , Óxido Nítrico Sintase Tipo III , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Coelhos , Resultado do Tratamento , Vasodilatadores/administração & dosagem , Vitamina E/uso terapêutico
15.
Diabetes Care ; 26(2): 285-9, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12547850

RESUMO

OBJECTIVE: We investigated the effect of glimepiride, a third-generation sulfonylurea hypoglycemic agent, on insulin resistance in elderly patients with type 2 diabetes, in connection with plasma adiponectin and 8-epi-prostagrandin F2alpha (8-epi-PGF2alpha), an oxidative stress marker. RESEARCH DESIGN AND METHODS: A total of 17 elderly patients with type 2 diabetes received 12 weeks of treatment with glimepiride. Homeostasis assessment model of insulin resistance (HOMA-IR), homeostasis assessment model of beta-cell function, HbA(1c), C-peptide in 24-h pooled urine (urine CPR), and plasma concentrations of 8-epi-PGF2alpha, tumor necrosis factor-alpha (TNF-alpha), plasminogen activator inhibitor type 1, and adiponectin were measured at various times. The metabolic clearance rate of glucose (MCR-g) was also assessed by a hyperinsulinemic-euglycemic clamp. RESULTS: After 8 weeks of glimepiride treatment, significant reductions were observed in HbA(1c) (from 8.4 +/- 1.9 to 6.9 +/- 1.0%), HOMA-IR (from 2.54 +/- 2.25 to 1.69 +/- 0.95%), and plasma TNF-alpha concentrations (from 4.0 +/- 2.0 to 2.6 +/- 2.5 pg/ml). MCR-g was significantly increased from 3.92 +/- 1.09 to 5.73 +/- 1.47 mg. kg(-1). min(-1). Plasma adiponectin increased from 6.61 +/- 3.06 to 10.2 +/- 7.14 micro g/ml. In control subjects, who maintained conventional treatment, no significant changes were observed in any of these markers. CONCLUSIONS: Glimepiride remarkably improved insulin resistance, suggested by a significant reduction in HOMA-IR, an increase in MCR-g, and a reduction in HbA(1c) without changing extrapancreatic beta-cell function and urine CPR. Increased plasma adiponectin and decreased plasma TNF-alpha may underlie the improvement of insulin resistance with glimepiride.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/fisiopatologia , Dinoprosta/análogos & derivados , Hipoglicemiantes/uso terapêutico , Resistência à Insulina , Peptídeos e Proteínas de Sinalização Intercelular , Proteínas/análise , Compostos de Sulfonilureia/uso terapêutico , Adiponectina , Idoso , Dinoprosta/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Concentração Osmolar , Inibidor 1 de Ativador de Plasminogênio/sangue , Fator de Necrose Tumoral alfa/análise
16.
Synapse ; 43(3): 195-200, 2002 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-11793425

RESUMO

For functional assessment of gene therapy in experimental animals, in vivo assessment of transferred genes will provide a major advance over an in vitro analysis which must be done post-hoc. In the current study we conducted positron emission tomography (PET) analysis in rats following injection of the adenoviral vector encoding the cDNA for the rat dopamine D(2) receptors (D(2)R) (AdCMV.DopD(2)R) into rat brain to provide a quantitative evaluation of D(2)R overexpression. Quantitative measurements as well as images by PET and ex vivo autoradiography demonstrated the significant increase of D(2)R binding of [(11)C]raclopride, a specific D(2)R radioligand, in the AdCMV.DopD(2)R-injected rat striatum 2 or 3 days after vector injection. Longitudinal in vivo assessment of the gene expression by PET demonstrated decreased binding of [(11)C]raclopride with time, which was in agreement with the observation in a cross-sectional autoradiographic study. The results of the current study demonstrate that PET can be used for longitudinal in vivo assessment of D(2)R expression mediated by adenoviral vector in rat brain.


Assuntos
Ligação Competitiva/fisiologia , Corpo Estriado/diagnóstico por imagem , Regulação da Expressão Gênica/fisiologia , Vetores Genéticos , Neurônios/metabolismo , Receptores de Dopamina D2/metabolismo , Adenoviridae/genética , Animais , Ligação Competitiva/efeitos dos fármacos , Corpo Estriado/efeitos dos fármacos , Antagonistas de Dopamina , Regulação da Expressão Gênica/efeitos dos fármacos , Terapia Genética/instrumentação , Terapia Genética/métodos , Masculino , Neurônios/efeitos dos fármacos , Racloprida , Ratos , Ratos Endogâmicos F344 , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/genética , Tomografia Computadorizada de Emissão
17.
J Cardiovasc Pharmacol ; 39(2): 298-309, 2002 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11791016

RESUMO

It is not completely understood whether nitric oxide donors and beta-adrenoceptor antagonists have anti-atherosclerotic effects. The anti-atherosclerotic effects of beta-adrenergic receptor antagonists and nitric oxide donors on severe atherosclerosis induced by cholesterol and inhibition of nitric oxide synthesis were determined. Six groups of New Zealand white male rabbits were treated for 10 weeks, under the following regimens: group I: high-cholesterol diet (HCD) (standard diet plus 0.5% cholesterol); group II: HCD plus N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase; group III: HCD plus L-NAME and isosorbide dinitrate; group IV: HCD plus L-NAME and nitroglycerin; group V: HCD plus L-NAME and nipradilol (beta-blocker with nitric oxide-releasing action); and group VI: HCD plus L-NAME and atenolol (beta-blocker). Serum lipid levels did not differ among the six groups. Blood pressure and heart rates were slightly decreased in groups V and VI. The atherosclerotic area and aortic cholesterol increased in L-NAME-treated animals but not in animals in group V. The endothelium-dependent relaxations and basal nitric oxide release were impaired in the L-NAME treatment group, though not in group V, in comparison with those in group I. cGMP in the aorta was increased in groups III, IV, and V as compared with that in group II. Endothelial nitric oxide synthase mRNA was decreased in the aortae of L-NAME-treated rabbits and increased in aortae in group V, in comparison with that in group I. Conclusively, nipradilol, beta-blocker with nitric oxide-releasing action, in contrast to the other beta-blockers and nitric oxide donors, showed a successful anti-atherosclerotic effect through the restoration of nitric oxide bioavailability and possible interaction with oxygen radicals.


Assuntos
Antagonistas Adrenérgicos beta/uso terapêutico , Arteriosclerose/tratamento farmacológico , Doadores de Óxido Nítrico/uso terapêutico , Propanolaminas/uso terapêutico , Antagonistas Adrenérgicos beta/farmacologia , Animais , Aorta Torácica/química , Aorta Torácica/metabolismo , Aorta Torácica/patologia , Arteriosclerose/etiologia , Atenolol/farmacologia , Atenolol/uso terapêutico , Pressão Sanguínea/efeitos dos fármacos , Colesterol/análise , Colesterol na Dieta/efeitos adversos , Endotélio Vascular/metabolismo , Endotélio Vascular/fisiopatologia , Frequência Cardíaca/efeitos dos fármacos , Dinitrato de Isossorbida/farmacologia , Dinitrato de Isossorbida/uso terapêutico , Masculino , Relaxamento Muscular/efeitos dos fármacos , Óxido Nítrico/deficiência , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/genética , Nitroglicerina/farmacologia , Nitroglicerina/uso terapêutico , Propanolaminas/farmacologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Coelhos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vasodilatadores/farmacologia , Vasodilatadores/uso terapêutico
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