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1.
Bone Joint J ; 101-B(5): 565-572, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31038991

RESUMO

AIMS: The purpose of the present study was to compare patient-specific instrumentation (PSI) and conventional surgical instrumentation (CSI) for total knee arthroplasty (TKA) in terms of early implant migration, alignment, surgical resources, patient outcomes, and costs. PATIENTS AND METHODS: The study was a prospective, randomized controlled trial of 50 patients undergoing TKA. There were 25 patients in each of the PSI and CSI groups. There were 12 male patients in the PSI group and seven male patients in the CSI group. The patients had a mean age of 69.0 years (sd 8.4) in the PSI group and 69.4 years (sd 8.4) in the CSI group. All patients received the same TKA implant. Intraoperative surgical resources and any surgical waste generated were recorded. Patients underwent radiostereometric analysis (RSA) studies to measure femoral and tibial component migration over two years. Outcome measures were recorded pre- and postoperatively. Overall costs were calculated for each group. RESULTS: There were no differences (p > 0.05) in any measurement of migration at two years for either the tibial or femoral components. Movement between one and two years was < 0.2 mm, indicating stable fixation. There were no differences in coronal or sagittal alignment between the two groups. The PSI group took a mean 6.1 minutes longer (p = 0.04) and used a mean 3.4 less trays (p < 0.0001). Total waste generated was similar (10 kg) between the two groups. The PSI group cost a mean CAD$1787 more per case (p < 0.01). CONCLUSION: RSA criteria suggest that both groups will have revision rates of approximately 3% at five years. The advantages of PSI were minimal or absent for surgical resources used and waste eliminated, and for meeting target alignment, yet had significantly greater costs. Therefore, we conclude that PSI may not offer any advantage over CSI for routine primary TKA cases. Cite this article: Bone Joint J 2019;101-B:565-572.


Assuntos
Artroplastia do Joelho/instrumentação , Assistência à Saúde/estatística & dados numéricos , Custos de Cuidados de Saúde/estatística & dados numéricos , Prótese do Joelho/efeitos adversos , Idoso , Artroplastia do Joelho/efeitos adversos , Artroplastia do Joelho/economia , Canadá , Assistência à Saúde/economia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/cirurgia , Estudos Prospectivos , Desenho de Prótese , Falha de Prótese , Resultado do Tratamento
2.
Horm Metab Res ; 36(4): 197-202, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15114516

RESUMO

Gender-related differences in cardiac function have been described in the literature, but whether the presence of sex hormones is responsible for these differences remains unclear. This study was designed to determine whether testosterone regulates the gene expression of calcium regulatory proteins in rat heart, thus playing a role in gender-related differences in cardiac performance. Ventricular myocytes were isolated from two-day-old rats and treated with testosterone at varying duration; the levels of gene expression for the androgen receptor (AR) and major calcium regulatory proteins were determined by quantitative real-time PCR. Testosterone (1 microM) treatment induced a maximum increase in beta1-adrenergic receptor and L-type calcium channel mRNA levels following an eight hour exposure. Six hours testosterone treatment stimulated a 300-fold increase in androgen receptor message abundance, and Na/Ca exchanger mRNA levels reached a maximum level following twenty-four hour testosterone treatment. Taken together, these data provide the first evidence that testosterone regulates gene expression of the major calcium regulatory proteins in isolated ventricular myocytes, and may thus play a role in the gender-related differences observed in cardiac performance.


Assuntos
Androgênios/farmacologia , Canais de Cálcio Tipo L/genética , Miócitos Cardíacos/efeitos dos fármacos , Receptores Adrenérgicos beta 1/genética , Testosterona/farmacologia , Animais , Células Cultivadas , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Miócitos Cardíacos/citologia , Miócitos Cardíacos/fisiologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Caracteres Sexuais , Trocador de Sódio e Cálcio/genética
3.
Am J Physiol Heart Circ Physiol ; 280(3): H1376-82, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11179087

RESUMO

The Na/Ca exchanger encoded by the NCX1 gene plays an important role in calcium homeostasis in cardiac muscle. We previously identified three in vitro signaling pathways that are of major importance in the regulation of Na/Ca exchanger gene expression in neonatal cardiac myocytes, the protein kinase A (PKA) and protein kinase C (PKC) pathways, and intracellular Ca(2+). To determine whether these pathways are important in vivo, we stimulated the PKA and PKC pathways and examined functional expression of the Na/Ca exchanger in adult rat heart. After a 3- and 7-day treatment, norepinephrine (200 microg x kg(-1) x h(-1)), isoproterenol (150 microg x kg(-1) x h(-1)), and phenylephrine (200 microg x kg(-1) x h(-1)) each stimulated a significant increase in NCX1 mRNA levels (35-85%, P < 0.05). Norepinephrine also stimulated a 35% increase in protein abundance (P < 0.05), a 20% decrease in relaxation duration (P < 0.05), and a 25% reduction in the fluorescence decay constant (P < 0.05) after a 7-day treatment. We conclude that a 7-day treatment of alpha- and beta-adrenergic agonists increases the expression of functional Na/Ca exchangers in adult rat heart.


Assuntos
Miocárdio/metabolismo , Receptores Adrenérgicos/metabolismo , Trocador de Sódio e Cálcio/genética , Trocador de Sódio e Cálcio/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/fisiologia , Pressão Sanguínea , Cálcio/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Expressão Gênica/fisiologia , Frequência Cardíaca , Isoproterenol/farmacologia , Masculino , Fibras Musculares Esqueléticas/enzimologia , Miocárdio/citologia , Norepinefrina/farmacologia , Fenilefrina/farmacologia , Proteína Quinase C/metabolismo , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley
4.
Cardiovasc Drugs Ther ; 14(5): 459-61, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11101192

RESUMO

The T-type calcium channel offers a new therapeutic target for treatment of patients with cardiovascular disease. Mibefradil, a T channel blocker, produces heart rate slowing and coronary vasodilatation but without the negative inotropic effect commonly seen when L-type channel blockers are used. The present study shows Mibefradil prevents ischemic episodes that are and are not preceded by an increase in heart rate. Although Mibefradil has been withdrawn because of multiple drug interactions, new T-type calcium channel blockers are under development.


Assuntos
Bloqueadores dos Canais de Cálcio/uso terapêutico , Mibefradil/uso terapêutico , Isquemia Miocárdica/tratamento farmacológico , Canais de Cálcio/efeitos dos fármacos , Doença Crônica , Interações de Medicamentos , Frequência Cardíaca/efeitos dos fármacos , Humanos , Isquemia Miocárdica/prevenção & controle
5.
J Mol Cell Cardiol ; 32(10): 1841-9, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11013128

RESUMO

The L-type calcium channel is a heteromultimeric protein complex, which is expressed in the cardiac sarcolemma. Although post-translational regulation of its subunits by protein kinase A (PKA) has been widely reported, little is known about molecular processes that regulate expression of calcium channel subunits (alpha(1C), alpha(2)- delta, and beta(2A)subunits). Previous studies from our group demonstrate that the steady-state mRNA level of the alpha(1C)unit is increased by treatment of myocytes with beta -adrenergic agonists. The current study is designed to determine whether the mRNA levels for all subunits of the L-type calcium channel are coordinately controlled by a beta -adrenergic agonist, and whether this occurs predominantly through control of rate of transcription. Nuclear run-on assays were used to determine the transcription initiation rate of these genes in cultured neonatal rat cardiac myocytes. In isoproterenol (10(-7)m)-treated myocytes, transcription of genes encoding the alpha(1C), alpha(2)- delta, and beta(2A)subunits was enhanced. The increases in transcription initiation rate for alpha(1C), alpha(2)- delta, and beta(2A)subunits genes were 404%, 367%, and 240% of control, respectively. Pretreatment with the beta -adrenergic antagonist propranolol (10(-5)m) or PKA inhibitor H-89 (10(-6)m) blocked the effects of isoproterenol, while either drug alone did not affect the gene transcription rate significantly. Steady state mRNA levels of the subunits increased following isoproterenol treatment. These results suggest that beta -adrenergic stimulation and the PKA signaling pathway play an important role in transcriptional regulation of the L-type calcium channel in myocyte. The expression of all the subunits of this ion channel is under coordinate transcriptional control.


Assuntos
Canais de Cálcio Tipo L/metabolismo , Regulação da Expressão Gênica , Sulfonamidas , Transcrição Genética , Agonistas Adrenérgicos beta/farmacologia , Animais , Animais Recém-Nascidos , Northern Blotting , Cardiotônicos/farmacologia , Núcleo Celular/metabolismo , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , DNA Complementar/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Ventrículos do Coração/metabolismo , Isoproterenol/farmacologia , Isoquinolinas/farmacologia , Miocárdio/citologia , Miocárdio/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Fatores de Tempo
7.
Mol Cell Biochem ; 205(1-2): 101-9, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10821427

RESUMO

We used Northern analyses, RNase protection assays and immunoblot analyses to examine the relationship among developmental age of the heart, abundance of mRNA and L-type calcium channel alpha1C subunit protein, and to establish the size of the native protein in heart. Northern analysis, RNase protection assays, and immunoblots were used to study RNA and protein from rat heart of various ages. In fetal and adult ventricles there was a predominant 8.3-kb transcript for the alpha1C subunit with no change in transcript size during development. RNase protection assays demonstrated a 2-fold increase in abundance of the DHP receptor message during postnatal development. Immunoblots identified a 240 kD protein, corresponding to the predicted molecular mass of the full length alpha1C subunit. No change in size of protein for the alpha1C subunit was observed at any developmental stage and there was no evidence for a truncated isoform. There was an approximate 2-fold increase in alpha1C subunit protein in ventricular homogenates during postnatal development. Thus, in the developing rat heart, alterations in calcium channel properties during development appear to result neither from alternative splicing that produces a smaller transcript for the alpha1C subunit nor from expression of a truncated protein, but at least in part from transcriptionally-regulated expression of the 240 kDa polypepde.


Assuntos
Canais de Cálcio Tipo L/biossíntese , Canais de Cálcio Tipo L/química , Miocárdio/metabolismo , Fatores Etários , Animais , Northern Blotting , Membrana Celular/metabolismo , Eletroforese em Gel de Poliacrilamida , Feminino , Immunoblotting , Masculino , Isoformas de Proteínas , RNA Antissenso/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Transcrição Genética
8.
J Mol Cell Cardiol ; 32(4): 611-20, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10757741

RESUMO

The Na/Ca exchanger protein encoded by the NCX1 gene provides the predominant mechanism for calcium efflux during cardiac relaxation. Because beta -adrenergic stimulation increases expression of Ca(2+)channels (Ca(2+)influx) in cardiac myocytes, we tested the hypothesis that isoproterenol would concomitantly augment expression of NCX1. Four hour treatment of neonatal myocytes with isoproterenol significantly increased NCX1 gene and protein expression, and increased the rate of transcript initiation. Alpha-adrenergic stimulation significantly decreases NCX1 mRNA levels. Calcium transient measurements revealed that for cells that had been pretreated with isoproterenol there was a faster relaxation rate of the Ca(2+)transient in the presence of thapsigargin, indicating an enhanced rate of intracellular Ca(2+)removal. We conclude that effectors that increase calcium channel expression in neonatal myocytes also augments NCX1 gene and protein expression over a similar time course, and that this is due to enhanced NCX1 transcription. The regulation of expression of NCX1 by adrenergic pathways may play an important role in regulation of excitation-contraction coupling in cardiac myocytes.


Assuntos
Ventrículos do Coração/metabolismo , Miocárdio/metabolismo , Receptores Adrenérgicos beta/metabolismo , Trocador de Sódio e Cálcio/genética , Agonistas alfa-Adrenérgicos/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Animais , Animais Recém-Nascidos , Cálcio/metabolismo , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Ventrículos do Coração/citologia , Isoproterenol/farmacologia , Miocárdio/citologia , Fenilefrina/farmacologia , RNA Mensageiro , Ratos , Ratos Sprague-Dawley , Transcrição Genética/efeitos dos fármacos
9.
Am J Physiol Heart Circ Physiol ; 278(4): H1153-62, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10749709

RESUMO

The alpha(1)-subunit of the cardiac/vascular Ca(2+) channel, which is the dihydropyridine (DHP)-binding site (the DHP receptor), provides the pore structure for Ca(2+) entry. It contains the binding sites for multiple classes of drugs collectively known as Ca(2+) antagonists. As an initial step toward understanding the mechanisms controlling transcription of the rat cardiac alpha(1C)-subunit gene, we have cloned a 2.3-kb fragment containing the 5'-flanking sequences and identified the alpha(1C)-subunit gene transcription start site. The rat alpha(1C)-subunit gene promoter belongs to the TATA-less class of such basal elements. Using deletion analysis of alpha(1C)-subunit promoter-luciferase reporter gene constructs, we have characterized the transcriptional modulating activity of the 5'-flanking region and conducted transient transfections in cultured neonatal rat cardiac ventricular myocytes and vascular smooth muscle cells. Sequence scanning identified several potential regulatory elements, including five consensus sequences for the cardiac-specific transcription factor Nkx2.5, an AP-1 site, a cAMP response element, and a hormone response element. Transient transfection experiments with the promoter-luciferase reporter fusion gene demonstrate that the 2-kb 5'-flanking region confers tissue specificity and hormone responsiveness to expression of the Ca(2+) channel alpha(1C)-subunit gene. Electrophoretic mobility shift assays identified a region of the alpha(1C)-subunit gene promoter that can bind transcription factors and appears to be important for gene expression.


Assuntos
Regiões 5' não Traduzidas/genética , Canais de Cálcio Tipo L/genética , Fibras Musculares Esqueléticas/fisiologia , Regiões Promotoras Genéticas/genética , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Agonistas alfa-Adrenérgicos/farmacologia , Agonistas Adrenérgicos beta/farmacologia , Animais , Sequência de Bases , Células Cultivadas , Eletroforese/métodos , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/genética , Genes Reporter , Teste de Complementação Genética , Hormônios Esteroides Gonadais/farmacologia , Técnicas In Vitro , Isoproterenol/farmacologia , Luciferases/genética , Dados de Sequência Molecular , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/citologia , Músculo Liso Vascular/química , Músculo Liso Vascular/citologia , Miocárdio/química , Miocárdio/citologia , Fenilefrina/farmacologia , RNA Mensageiro/metabolismo , Ratos , Testosterona/farmacologia , Transcrição Genética/efeitos dos fármacos , Transcrição Genética/fisiologia , Transfecção
10.
Am J Med ; 107(3): 254-61, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10492319

RESUMO

Myocardial infarction in persons under the age of 45 years accounts for 6% to 10% of all myocardial infarctions in the United States. In this age group, it is predominantly a disease of men. Important risk factors include a family history of myocardial infarction before age 55 years, hyperlipidemia, smoking, and obesity. Unlike older patients, approximately half of young patients have single-vessel coronary disease, and in up to 20%, the cause is not related to atherosclerosis. Coronary angiography may be warranted in young patients with myocardial infarction to define the anatomy of the disease and to permit optimal management.


Assuntos
Infarto do Miocárdio , Algoritmos , Angioplastia Coronária com Balão , Ponte de Artéria Coronária , Árvores de Decisões , Diagnóstico Diferencial , Humanos , Incidência , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/epidemiologia , Infarto do Miocárdio/etiologia , Infarto do Miocárdio/terapia , Prognóstico , Fatores de Risco , Terapia Trombolítica
11.
Am J Physiol ; 276(4 Pt 2): H1393-8, 1999 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10199867

RESUMO

Bradykinin (BK) has a direct hypertrophic effect on rat ventricular cardiomyocytes (VCM) as defined by an increase in protein synthesis and an increase in atrial natriuretic peptide mRNA and secretion. In the current study, we have examined the dependence of BK-induced protein synthesis on activation of 90-kDa ribosomal S6 kinase (p90(rsk)) and 70-kDa S6 kinase (p70(S6K)). Both of these kinases possess the ability to phosphorylate the ribosomal protein S6, which plays an important role in initiating mRNA translation. Stimulation of adult VCM with 10 microM BK increased p90(rsk) activity by 2.5 +/- 0.3-fold and increased p70(S6K) activity by 2.0 +/- 0.3-fold. p90(rsk) is a terminal kinase in the mitogen-activated protein (MAP) kinase pathway. Inhibition of MAP kinase kinase activation by Raf in the MAP kinase pathway with PD-098059 (25 microM) blocked BK-stimulated activation of p90(rsk) by 70% and unexpectedly blocked p70(S6K) by 72%. Rapamycin inhibited BK-stimulated p70(S6K) activity by 93% but had no effect on p90(rsk) activation by BK. Inhibition of the MAP kinase pathway and p70(S6K) with PD-098059 was paralleled by changes in protein synthesis. BK (10 microM) increased [3H]phenylalanine incorporation by 27 +/- 3 and 39 +/- 6% in cultured adult and neonatal VCM, respectively. Treatment with PD-098059 or rapamycin abolished the increase in protein synthesis stimulated by BK. These results suggest that 1) BK activates p70(S6K) and p90(rsk); 2) although both p70(S6K) and p90(rsk) have the potential to phosphorylate the ribosomal S6 protein, p70(S6K) and not p90(rsk) is the predominant kinase involved in increasing protein synthesis by BK; and 3) p70(S6K) activation is dependent on stimulation of the MAP kinase pathway at a point distal to Raf.


Assuntos
Bradicinina/farmacologia , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas Musculares/biossíntese , Miocárdio/metabolismo , Proteínas Quinases S6 Ribossômicas/fisiologia , Angiotensina II/farmacologia , Animais , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Masculino , Proteínas Musculares/antagonistas & inibidores , Miocárdio/citologia , Fenilalanina/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Quinases S6 Ribossômicas/antagonistas & inibidores , Proteínas Quinases S6 Ribossômicas/metabolismo , Sirolimo/farmacologia
12.
Am J Physiol ; 275(4 Pt 2): H1370-4, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9746487

RESUMO

The aim of the present study was to test the hypothesis that bradykinin-stimulated release of nitric oxide (NO) and/or prostacyclin from endothelium blocks myocyte hypertrophy in vitro. Angiotensin II increased [3H]phenylalanine incorporation by 21 +/- 2% in myocytes cocultured with endothelial cells; this was abolished by bradykinin in the presence of endothelial cells. Bradykinin increased cytosolic concentrations of cGMP by 29 +/- 4% in myocytes cocultured with endothelial cells. This was abolished by inhibition of NO synthase and by a cyclooxygenase inhibitor. Angiotensin II also increased [3H]phenylalanine incorporation by 28 +/- 3% in myocytes cultured in the absence of endothelial cells. This effect of angiotensin II in monoculture was abolished by donors of NO but not by bradykinin. Neither the stable analog of prostacyclin (iloprost) nor the prostacyclin second messanger analog 8-bromo-cAMP (8-BrcAMP) blocked the effect of angiotensin II. Furthermore, 8-BrcAMP and iloprost individually increased [3H]phenylalanine incorporation. The antihypertrophic effects of bradykinin are critically dependent on endothelium-derived NO.


Assuntos
Angiotensina II/farmacologia , Cardiomegalia , Coração/fisiologia , Miocárdio/metabolismo , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico/fisiologia , 1-Metil-3-Isobutilxantina/farmacologia , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Angiotensina II/fisiologia , Animais , Bradicinina/farmacologia , Células Cultivadas , Técnicas de Cocultura , GMP Cíclico/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/fisiologia , Coração/efeitos dos fármacos , Iloprosta/farmacologia , Indometacina/farmacologia , Masculino , Molsidomina/análogos & derivados , Molsidomina/farmacologia , Miocárdio/citologia , Nitroprussiato/farmacologia , Fenilalanina/metabolismo , Biossíntese de Proteínas , Ratos , Ratos Sprague-Dawley , ômega-N-Metilarginina/farmacologia
13.
Circulation ; 98(3): 256-61, 1998 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-9697826

RESUMO

BACKGROUND: The role of androgens in producing cardiac hypertrophy by direct action on cardiac myocytes is uncertain. Accordingly, we tested the hypothesis that cardiac myocytes in adult men and women express an androgen receptor gene and that myocytes respond to androgens by a hypertrophic response. METHODS AND RESULTS: We used reverse transcription-polymerase chain reaction methods to demonstrate androgen receptor transcripts in multiple tissues and [3H]phenylalanine incorporation and atrial natriuretic peptide secretion as markers of hypertrophy in cultured rat myocytes. Messenger RNA encoding androgen receptors was detected in myocytes of male and female adult rats, neonatal rat myocytes, rat heart, dog heart, and infant and adult human heart. Both testosterone and dihydrotestosterone produced a robust receptor-specific hypertrophic response in myocytes, determined by indices of protein synthesis and atrial natriuretic peptide secretion. CONCLUSIONS: Androgen receptors are present in cardiac myocytes from multiple species, including normal men and women, in a context that permits androgens to modulate the cardiac phenotype and produce hypertrophy by direct, receptor-specific mechanisms. There are clinical implications for therapeutic or illicit use of androgens in humans.


Assuntos
Cardiomegalia/etiologia , Miocárdio/metabolismo , Receptores Androgênicos/fisiologia , Adulto , Animais , Células Cultivadas , Cães , Feminino , Humanos , Lactente , Masculino , Miocárdio/patologia , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Androgênicos/genética , Transdução de Sinais/fisiologia , Transcrição Genética
14.
Hypertension ; 31(1): 39-44, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9449388

RESUMO

Angiotensin-converting enzyme inhibitors block left ventricular hypertrophy in vivo. A component of this effect has been attributed to tissue accumulation of bradykinin. Little is known regarding the effect of bradykinin on cardiomyocytes. The objectives of the present study were to define the effects of bradykinin on isolated ventricular cardiomyocytes (from adult and neonatal rat hearts) and to determine the extent to which bradykinin blocks hypertrophy in vitro. Bradykinin was found to be a hypertrophic agonist, as defined by increased protein synthesis and atrial natriuretic peptide secretion and expression. Bradykinin (10 micromol/L) increased [3H]phenylalanine incorporation by 23+/-3% in adult and by 36+/-10% in neonatal cardiomyocytes. Constitutive atrial natriuretic peptide secretion by neonatal myocytes was increased 357+/-103%. All effects of bradykinin were abolished by the B2-kinin receptor antagonist Hoe 140. These increases were similar in magnitude to those observed with phenylephrine (20 micromol/L) and angiotensin II (1 micromol/L). However, in cardiomyocytes cocultured with endothelial cells, bradykinin did not increase protein synthesis. Angiotensin II increased [3H]phenylalanine incorporation by 24+/-3% in adult cardiomyocytes in monoculture and by 22+/-2% in adult rat cardiomyocytes cocultured with endothelial cells. Bradykinin abolished this angiotensin II-induced hypertrophy in myocytes cultured with endothelial cells but not in myocytes studied in the absence of endothelial cells. In conclusion, bradykinin has a direct hypertrophic effect on ventricular myocytes. The presence of endothelial cells is required for the antihypertrophic effects of bradykinin. The results suggest that the increase in local concentration of bradykinin associated with angiotensin-converting enzyme inhibition is an important mechanism by which hypertrophy can be blocked. Manifestation of this mechanism appears to require bradykinin-stimulated release of paracrine factor(s) from endothelial cells, which are also able to block the hypertrophic effects of Ang II.


Assuntos
Bradicinina/farmacologia , Hipertrofia Ventricular Esquerda/induzido quimicamente , Angiotensina II/farmacologia , Animais , Animais Recém-Nascidos , Bradicinina/análogos & derivados , Antagonistas dos Receptores da Bradicinina , Células Cultivadas , Endotélio/citologia , Ventrículos do Coração/citologia , Ventrículos do Coração/efeitos dos fármacos , Hipertrofia Ventricular Esquerda/prevenção & controle , Masculino , Ratos , Ratos Sprague-Dawley
15.
J Mol Cell Cardiol ; 29(7): 1791-803, 1997 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9236134

RESUMO

In addition to playing a significant role in cardiac excitation-contraction coupling, intracellular Ca2+ ([Ca2+]i) can regulate gene expression. While the mechanisms regulating expression of Ca2+ channels are not entirely defined, some evidence exists for Ca2+-dependent regulation. Using an adult ventricular myocyte culture system, we determined the effects of Ca2+ on: (1) abundance of mRNA for L-type Ca2+ channel alpha1 subunit (DHP receptor); (2) amount of DHP receptors; and (3) whole-cell Ca2+ current (ICa). Rat ventricular myocytes were cultured for 1-3 days in serum-free medium containing either normal (1.8 mM) or high (4.8 mM) Ca2+. Exposing myocytes to high Ca2+ rapidly elevated [Ca2+]i as determined by fura-2. Northern blot analysis revealed that culturing cells in high Ca2+ produced 1.5-fold increase in mRNA levels for the DHP receptor. The abundance of DHP receptors, determined by ligand binding, was two-fold greater in myocytes after 3 days in high Ca2+. Moreover, peak ICa was larger in myocytes cultured for 3 days in high Ca2+ (-17.8+/-1.5 pA/pF, n=26) than in control cells (-11.0+/-1.0 pA/pF, n=23). Voltage-dependent activation and inactivation, rates of current decay, as well as percent increases in ICa elicited by Bay K8644 were similar in all groups. Therefore, larger ICa is likely to represent a greater number of functional channels with unchanged kinetics. Our data support the conclusion that transient changes in [Ca2+]i can modulate DHP receptor mRNA and protein abundance, producing a corresponding change in functional Ca2+ channels in adult ventricular myocytes.


Assuntos
Canais de Cálcio/metabolismo , Cálcio/fisiologia , Miocárdio/metabolismo , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio Tipo L , Células Cultivadas , Regulação da Expressão Gênica , Ativação do Canal Iônico , Masculino , Proteínas Musculares/metabolismo , Técnicas de Patch-Clamp , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley
17.
J Clin Invest ; 97(3): 656-63, 1996 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-8609220

RESUMO

Expression of the dihydropyridine (DHP) receptor (alpha 1 subunit of L-type calcium channel) in heart is regulated by differentiation and innervation and is altered in congestive heart failure. We examined the transmembrane signaling pathways by which norepinephrine regulates DHP receptor expression in cultured neonatal rat ventricular myocytes. Using a 1.3-kb rat cardiac DHP receptor probe, and Northern analysis quantified by laser densitometry, we found that norepinephrine exposure produced a 2.2-fold increase in DHP receptor mRNA levels at 2 h followed by a decline to 50% of control at 4-48 h (P < 0.02). The alpha-adrenergic agonist phenylephrine and a phorbol ester produced a decline in mRNA levels (8-48 h). The beta-adrenergic agonist isoproterenol and 8-bromo-cAMP produced a transient increase in mRNA levels. After 24 h of exposure to isoproterenol, 3H-(+)PN200-110 binding sites increased from 410 +/- 8 to 539 +/- 39 fmol/mg (P < 0.05). The number of functional calcium channels, estimated by whole-cell voltage clamp experiments, was also increased after 24 h of exposure to isoproterenol. Peak current density (recordings performed in absence of isoproterenol) increased from -10.8 +/- 0.8 (n = 23) to -13.9 +/- 1.0 pA/pF (n = 27) (P < 0.01). Other characteristics of the calcium current (voltage for peak current, activation, and inactivation) were unchanged. Exposure for 48 h to phenylephrine produced a significant decline in peak current density (P < 0.01). We conclude that beta -adrenergic transmembrane signaling increases DHP receptor mRNA and number of functional calcium channels and that alpha - adrenergic transmembrane signaling produces a reciprocal effect. Regulation of cardiac calcium channel expression by adrenergic pathways may have physiological and pathophysiological importance.


Assuntos
Canais de Cálcio/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Ventrículos do Coração/metabolismo , Proteínas Musculares/biossíntese , Miocárdio/metabolismo , Receptores Colinérgicos/biossíntese , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Animais Recém-Nascidos , Canais de Cálcio/genética , Canais de Cálcio Tipo L , Catecolaminas/farmacologia , Células Cultivadas , Eletrofisiologia , Ventrículos do Coração/citologia , Proteínas Musculares/genética , Miocárdio/citologia , Norepinefrina/farmacologia , Fenilefrina/farmacologia , RNA Mensageiro/análise , Ratos , Receptores Colinérgicos/genética
18.
Avian Dis ; 39(4): 743-51, 1995 Oct-Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8719208

RESUMO

The effect of serotype 2 and 3 Marek's disease virus (MDV) vaccines on the development of pre-neoplastic bursal lesions induced by two strains of subgroup A avian leukosis virus (ALV) was studied. Chickens of line 15I5 x 7(1) susceptible to ALV-induced lymphoma were inoculated at hatch with Rous-associated virus-1 (RAV-1) or strain RPL-40 of subgroup A ALV. All chickens, except for negative controls, were also inoculated at hatch with strain 301B/1 of serotype 2 MDV or strain FC126 of turkey herpesvirus (HVT), a serotype 3 MDV. At 11 and 14 weeks of age, serial sections of bursal tissues from chickens in various treatment groups were stained with methyl green pyronin and examined for ALV-induced pre-neoplastic bursal lesions, also known as hyperplastic follicles. At 6 days and at 11 and 14 weeks of age, bursal tissues from groups of chickens inoculated with serotype 2 MDV and ALV at hatch were also examined for the presence of MDV genome by in situ hybridization. The number of ALV-induced hyperplastic follicles was significantly higher in chickens inoculated with serotype 2 MDV than in unvaccinated chickens or in chickens vaccinated with HVT. In contrast, the lowest number of hyperplastic follicles was noted in chickens vaccinated with HVT, regardless of strain of ALV used. These results suggest that enhancement of lymphoid leukosis may result from an increase in the number of ALV-induced hyperplastic follicles caused by the MDV and also that the enhancing effect of MDV on ALV-induced lymphomagenesis may occur at the stage of formation of hyperplastic follicles in the bursa.


Assuntos
Vírus da Leucose Aviária , Leucose Aviária/imunologia , Bolsa de Fabricius/patologia , Herpesvirus Galináceo 2/imunologia , Lesões Pré-Cancerosas/imunologia , Vacinas Virais , Animais , Leucose Aviária/patologia , Embrião de Galinha , Galinhas , Suscetibilidade a Doenças , Herpesvirus Galináceo 2/classificação , Lesões Pré-Cancerosas/patologia , Sorotipagem , Fatores de Tempo
19.
J Clin Invest ; 94(4): 1470-6, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7929822

RESUMO

Transforming growth factor-beta 1 (TGF-beta 1) is a peptide growth factor that may play a role in the myocardial response to hypertrophic stimuli. However, the cellular distribution, mechanism of induction, and source of increased TGF-beta 1 in response to hypertrophic stimuli are not known. We tested the hypothesis that the cardiac myocyte responds to hypertrophic stimuli with the increased expression of TGF-beta 1. In adult rat ventricular myocardium freshly dissociated into myocyte and nonmyocyte cellular fractions, the preponderance of TGF-beta 1 mRNA visualized by Northern hybridization was in the nonmyocyte fraction. Abdominal aortic constriction (7 d) and subcutaneous norepinephrine infusion (36 h) each caused ventricular hypertrophy associated with 3.1-fold and 3.8-fold increases, respectively, in TGF-beta 1 mRNA in the myocyte fraction, but had no effect on the level of TGF-beta 1 mRNA in the nonmyocyte fraction. In ventricular myocytes, norepinephrine likewise caused a 4.1-fold increase in TGF-beta 1 mRNA associated with an increase in TGF-beta bioactivity. This induction of TGF-beta 1 mRNA occurred at norepinephrine concentrations as low as 1 nM and was blocked by prazosin, but not propranolol. NE did not increase the TGF-beta 1 mRNA level in nonmyocytes, primarily fibroblasts, cultured from neonatal rat ventricle. Thus, the cardiac myocyte responds to two hypertrophic stimuli, pressure overload and norepinephrine, with the induction of TGF-beta 1. These data support the view that TGF-beta 1, released by myocytes and acting in an autocrine and/or paracrine manner, is involved in myocardial remodeling by hypertrophic stimuli.


Assuntos
Cardiomegalia/metabolismo , Miocárdio/metabolismo , Norepinefrina/farmacologia , Fator de Crescimento Transformador beta/biossíntese , Animais , Animais Recém-Nascidos , Aorta Abdominal , Fator Natriurético Atrial/biossíntese , Fator Natriurético Atrial/genética , Cardiomegalia/etiologia , Células Cultivadas , Constrição Patológica , Meios de Cultivo Condicionados/farmacologia , Fibroblastos/metabolismo , Ventrículos do Coração , Masculino , Miocárdio/citologia , Prazosina/farmacologia , Propranolol/farmacologia , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta/genética , Regulação para Cima/efeitos dos fármacos
20.
J Mol Cell Cardiol ; 26(9): 1187-95, 1994 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7815461

RESUMO

The ability of acadesine to modulate injury in a myocyte model of simulated ischemia was studied. When freshly isolated adult rat ventricular myocytes were subjected to 10 min of simulated ischemia and reperfusion, greater that 75% of myocytes developed hypercontracture and the amplitude of contraction of the remaining, potentially viable myocytes was markedly depressed. When cells were pretreated with 50 microM acadesine for 5 min and exposed to acadesine during simulated ischemia and during 2-10 min of reperfusion, followed by reperfusion with control buffer, up to 90% of myocytes maintained normal morphology and could be stimulated to contract. Acadesine alone had no significant effect on amplitude of contraction of the myocytes. Acadesine is known to alter adenosine metabolism and to increase coronary sinus [adenosine] in the ischemic heart. When cells were treated with the non-selective adenosine receptor antagonist 8-sulfonylphenyltheophylline, the protective effect of acadesine was abolished. Thus, in this adult rat cardiac myocyte model of simulated ischemia, acadesine protects partially against ischemia/reperfusion injury. The protective effect of acadesine is mediated, at least in part, by an adenosine receptor-dependent mechanism.


Assuntos
Aminoimidazol Carboxamida/análogos & derivados , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Ribonucleosídeos/farmacologia , Aminoimidazol Carboxamida/farmacologia , Animais , Modelos Animais de Doenças , Feminino , Coração/efeitos dos fármacos , Coração/fisiopatologia , Técnicas In Vitro , Contração Miocárdica/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Antagonistas de Receptores Purinérgicos P1 , Ratos , Ratos Sprague-Dawley , Teofilina/análogos & derivados , Teofilina/farmacologia
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