RESUMO
The gastrointestinal tract regulates glucose and energy metabolism, and there is increasing recognition that bile acids function as key signalling molecules in these processes. For example, bile acid changes that occur after bariatric surgery have been implicated in the effects on satiety, lipid and cholesterol regulation, glucose and energy metabolism, and the gut microbiome. In recent years, Takeda-G-protein-receptor-5 (TGR5), a bile acid receptor found in widely dispersed tissues, has been the target of significant drug discovery efforts in the hope of identifying effective treatments for metabolic diseases including type 2 diabetes, obesity, atherosclerosis, fatty liver disease and cancer. Although the benefits of targeting the TGR5 receptor are potentially great, drug development work to date has identified risks that include histopathological changes, tumorigenesis, gender differences, and questions about the translation of animal data to humans. The present article reviews the noteworthy challenges that must be addressed along the path of development of a safe and effective TGR5 agonist therapy.
Assuntos
Fármacos Antiobesidade/uso terapêutico , Diabetes Mellitus Tipo 2/tratamento farmacológico , Drogas em Investigação/uso terapêutico , Hipoglicemiantes/uso terapêutico , Modelos Biológicos , Obesidade/tratamento farmacológico , Receptores Acoplados a Proteínas G/agonistas , Animais , Fármacos Antiobesidade/efeitos adversos , Fármacos Antiobesidade/farmacologia , Diabetes Mellitus Tipo 2/metabolismo , Desenho de Fármacos , Descoberta de Drogas/tendências , Avaliação Pré-Clínica de Medicamentos , Drogas em Investigação/efeitos adversos , Drogas em Investigação/farmacologia , Humanos , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/farmacologia , Terapia de Alvo Molecular , Obesidade/metabolismo , Especificidade de Órgãos , Receptores Acoplados a Proteínas G/metabolismo , Pesquisa Translacional Biomédica/tendênciasRESUMO
AIMS: To investigate the pharmacodynamics, pharmacokinetics and safety/tolerability of blocking reuptake of bile acids using the inhibitor GSK2330672 (GSK672) in patients with type 2 diabetes (T2D). METHODS: Subjects with T2D taking metformin were enrolled in two studies in which they took metformin 850 mg twice daily for 2 weeks prior to and during the randomized treatment periods. In the first crossover study (n = 15), subjects received GSK672 45 mg, escalating to 90 mg, twice daily, or placebo for 7 days. The second parallel-group study (n = 75) investigated GSK672 10-90 mg twice daily, placebo or sitagliptin for 14 days. RESULTS: In both studies, GSK672 reduced circulating bile acids and increased serum 7-α-hydroxy-4-cholesten-3-one (C4), an intermediate in the hepatic synthesis of bile acids. Compared with placebo, in the parallel-group study 90 mg GSK672 twice daily reduced fasting plasma glucose [FPG; -1.21 mmol/l; 95% confidence interval (CI) -2.14, -0.28] and weighted-mean glucose area under the curve (AUC)0-24 h (-1.33 mmol/l; 95% CI -2.30, -0.36), as well as fasting and weighted-mean insulin AUC0 -24 h . GSK672 also reduced cholesterol (LDL, non-HDL and total cholesterol) and apolipoprotein B concentrations; the maximum LDL cholesterol reduction was â¼40%. There was no change in HDL cholesterol but there was a trend towards increased fasting triglyceride levels in the GSK672 groups compared with placebo. In both studies, the most common adverse events associated with GSK672 were gastrointestinal, mostly diarrhoea (22-100%), which appeared to be independent of dose. CONCLUSIONS: In subjects with T2D on metformin, GSK672 improved glucose and lipids, but there was a high incidence of gastrointestinal adverse events.
Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Hipoglicemiantes/administração & dosagem , Metformina/administração & dosagem , Metilaminas/administração & dosagem , Tiazepinas/administração & dosagem , Adulto , Apolipoproteínas B/metabolismo , Área Sob a Curva , Ácidos e Sais Biliares/metabolismo , Glicemia/metabolismo , LDL-Colesterol , Estudos Cross-Over , Diarreia/induzido quimicamente , Método Duplo-Cego , Esquema de Medicação , Jejum/metabolismo , Feminino , Gastroenteropatias/induzido quimicamente , Humanos , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/farmacologia , Insulina/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Metformina/efeitos adversos , Metformina/farmacologia , Metilaminas/efeitos adversos , Metilaminas/farmacologia , Pessoa de Meia-Idade , Transportadores de Ânions Orgânicos Dependentes de Sódio/antagonistas & inibidores , Simportadores/antagonistas & inibidores , Tiazepinas/efeitos adversos , Tiazepinas/farmacologia , Resultado do TratamentoRESUMO
We investigated the effects of a long-duration glucagon-like peptide-1 (GLP-1) receptor agonist, GSK2374697, on postprandial endogenous total GLP-1 and peptide YY (PYY). Two cohorts of healthy subjects, one normal/overweight and one obese, were randomized to receive GSK2374697 2 mg (n = 8 each) or placebo (n = 4 and n = 2) subcutaneously on days 1, 4 and 7. Samples for plasma endogenous GLP-1 and PYY were collected after breakfast on days -1 and 12. Weighted mean area under the curve (0-4 h) of total GLP-1 and PYY in treated subjects was reduced compared with placebo. The least squares mean difference for change from baseline was -1.24 pmol/l [95% confidence interval (CI) -2.33, -0.16] and -4.47 pmol/l (95% CI -8.74, -0.20) for total GLP-1 and PYY, respectively, in normal/overweight subjects (p < 0.05 for both), and -1.56 (95% CI -2.95, -0.16) and -3.02 (95% CI -8.58, 2.55), respectively, in obese subjects (p < 0.05 for GLP-1). In healthy subjects, GSK2374697 reduced postprandial total GLP-1 and PYY levels, suggesting feedback suppression of enteroendocrine L-cell secretion of these peptides.
Assuntos
Peptídeo 1 Semelhante ao Glucagon/efeitos dos fármacos , Peptídeo YY/efeitos dos fármacos , Período Pós-Prandial/efeitos dos fármacos , Proteínas Recombinantes de Fusão/farmacologia , Adulto , Idoso , Desjejum , Feminino , Peptídeo 1 Semelhante ao Glucagon/sangue , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeo YY/sangue , Adulto JovemRESUMO
Glucose-dependent insulinotropic polypeptide (GIP) communicates nutrient intake from the gut to islets, enabling optimal levels of insulin secretion via the GIP receptor (GIPR) on ß cells. The GIPR is also expressed in α cells, and GIP stimulates glucagon secretion; however, the role of this action in the postprandial state is unknown. Here, we demonstrate that GIP potentiates amino acid-stimulated glucagon secretion, documenting a similar nutrient-dependent action to that described in ß cells. Moreover, we demonstrate that GIP activity in α cells contributes to insulin secretion by invoking paracrine α to ß cell communication. Last, specific loss of GIPR activity in α cells prevents glucagon secretion in response to a meal stimulus, limiting insulin secretion and driving glucose intolerance. Together, these data uncover an important axis by which GIPR activity in α cells is necessary to coordinate the optimal level of both glucagon and insulin secretion to maintain postprandial homeostasis.
Assuntos
Diabetes Mellitus Tipo 2 , Incretinas , Polipeptídeo Inibidor Gástrico , Glucagon , Glucose , Humanos , Receptores Acoplados a Proteínas G , Receptores dos Hormônios GastrointestinaisRESUMO
Functional studies indicate that atrial natriuretic peptide (ANP), a member of the natriuretic peptide family, has direct effects on cardiac muscle cells. However, conventional ligand-binding studies designed to establish the presence of natriuretic peptide-binding sites in the heart have yielded conflicting results. There are discrepancies also between the latter and the receptor distribution predicted from the pattern of the mRNA transcripts localized by in situ hybridization. Here we have employed the technique of cDNA amplification with the polymerase chain reaction to confirm the presence of natriuretic peptide A, B, and C receptor mRNAs in rat and human cardiac tissue. In the rat heart, the distribution of the A and B receptor transcripts appears to be relatively homogeneous; in contrast, the C type mRNA is concentrated principally in the atria, with no difference between the left and right sides of the heart. A and B receptor DNA products were obtained after amplification of left, but not right, ventricular cDNA from the heart of a 16-yr-old male with cystic fibrosis; the yield of C receptor DNA was similar for both ventricles. If these mRNA transcripts are translated into functional receptors in the rat and human heart, ANP and the other natriuretic peptides may have direct effects on cardiac function, including regulation of natriuretic peptide release via a short feedback loop, modulation of contractility of the heart, or activation of cardiac reflexes.
Assuntos
Miocárdio/metabolismo , RNA Mensageiro/análise , Receptores do Fator Natriurético Atrial/genética , Animais , Fator Natriurético Atrial/metabolismo , Fator Natriurético Atrial/farmacologia , Sequência de Bases , Coração/efeitos dos fármacos , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Ratos , Ratos WistarRESUMO
The heart expresses the three natriuretic peptide receptors (NPR), namely NPR-A, NPR-B, and NPR-C. We have examined the temporal relationship between the expression of mRNA transcripts for atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) and their receptors in the heart during the development of cardiac hypertrophy in the aortovenocaval fistula rat. Messenger RNAs were measured by cDNA amplification. Progressive cardiac hypertrophy was accompanied by increased ANP mRNA prevalence throughout the heart and increased BNP mRNA in the left atrium. The most striking observation was the gradual disappearance of NPR-C transcripts (the putative "clearance" receptor) in all chambers; this was in marked contrast to the increase in mRNA levels for NPR-A and NPR-B (the guanylyl cyclase-linked receptors). Our observations have important therapeutic implications if the transcript changes are mirrored at the receptor protein level because (a) the apparent down-regulation of NPR-C may enhance the local action of natriuretic peptides on the heart, and (b) the loss of NPR-C, particularly if it is widespread, may reduce the rate of elimination of the natriuretic peptides, restricting the therapeutic potential of specific NPR-C ligands designed to reduce peptide clearance.
Assuntos
Cardiomegalia/metabolismo , Regulação da Expressão Gênica , Miocárdio/metabolismo , RNA Mensageiro/metabolismo , Receptores do Fator Natriurético Atrial/biossíntese , Animais , Sequência de Bases , Cardiomegalia/fisiopatologia , Primers do DNA , DNA Complementar/metabolismo , Guanilato Ciclase/metabolismo , Masculino , Dados de Sequência Molecular , Oligonucleotídeos Antissenso , Reação em Cadeia da Polimerase/métodos , Polimorfismo Genético , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Valores de Referência , Fatores de TempoRESUMO
Endothelin-1 (ET-1) is a potent vasoconstrictor peptide isolated from porcine endothelial cells. We have previously demonstrated widespread binding sites for ET-1 in blood vessels, heart, kidney, adrenal, lung, and brain in a distribution that paralleled that of endothelial cells. To determine whether these cells are capable of synthesizing ET-1 in close proximity to its binding sites, amplification of cDNA using the polymerase chain reaction and in situ hybridization were used to study the distribution of ET-1 mRNA. We have found widespread transcription of ET-1 mRNA in human and porcine tissues. The identity of the transcripts was confirmed by prediction of restriction fragment lengths or sequencing. In situ hybridization in the kidney showed that the regional expression of these transcripts is localized, probably to small blood vessels, but the failure to visualize ET-1 mRNA in the capillaries may reflect absence of expression or insufficient sensitivity of the technique. These results should permit investigation of the role of ET-1 as a local factor in vascular pathophysiology.
Assuntos
Peptídeos/genética , RNA Mensageiro/genética , Animais , Sequência de Bases , DNA/genética , Endotelinas , Endotélio Vascular/fisiologia , Humanos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos , Especificidade de Órgãos , Reação em Cadeia da Polimerase , RNA/isolamento & purificação , Mapeamento por Restrição , SuínosRESUMO
Resonance Raman spectra of cytochrome b6f complexes isolated from spinach chloroplasts have been obtained. Selective resonance enhancements and partial reductions of the complex by redox mediators were used to isolate and identify the contributions of heme b6 and heme f sites to the observed spectra. Corresponding spectra for turnip cytochrome f have also been obtained. Power-dependent photoreduction was observed in cytochrome f of the complex as well as in the isolated cytochrome f during the course of the Raman experiments.
Assuntos
Citocromos/química , Heme/química , Proteínas de Plantas/química , Citocromos f , Oxirredução , Análise Espectral RamanRESUMO
BACKGROUND: Conventional and tissue Doppler echocardiographically derived myocardial velocity gradients (MVGs) were used to characterize the myocardium in patients with Friedreich's ataxia (FRDA), and the relationship between MVGs and the mutation in the FRDA gene, a GAA triplet repeat expansion, was investigated. METHODS AND RESULTS: We studied 29 patients with FRDA (10 men, mean age 31+/-9 years) who were homozygous for the GAA expansion in the FRDA gene and were without cardiac symptoms. A comparison was made with a group of 30 age-matched control subjects. In patients with FRDA, interventricular septal thickness (1.17+/-0.26 versus 0.85+/-0.13 cm, P:<0.005), posterior left ventricular wall thickness (1.00+/-0.24 versus 0.88+/-0.15 cm, P:<0.01), and left atrial diameter (3.3+/-0.5 versus 2.9+/-0.3 cm, P:=0.01) were increased compared with control subjects. MVGs were reduced in FRDA during systole (3.1+/-1.2 versus 4.5+/-0.5 s(-1), P:<0.0001) and in early diastole (4.9+/-2.7 versus 8.8+/-1.8 s(-1), P:<0.0001) but increased in late diastole (2.0+/-1. 3 versus 1.1+/-0.9 s(-1), P:<0.01). The strongest relationship was seen between age-corrected early diastolic MVGs and the GAA expansion in the smaller allele of the FRDA gene (r=-0.68, P:<0. 0001). CONCLUSIONS: MVGs offer a means of further characterizing the myocardial abnormalities in patients with FRDA. Early diastolic MVGs appear to relate most closely to the genetic abnormality and the consequential reduction in frataxin protein.
Assuntos
Ataxia de Friedreich/patologia , Proteínas Adaptadoras de Transdução de Sinal , Adolescente , Adulto , Criança , Pré-Escolar , Ecocardiografia Doppler , Feminino , Ataxia de Friedreich/diagnóstico por imagem , Ataxia de Friedreich/genética , Ataxia de Friedreich/fisiopatologia , Genótipo , Humanos , Masculino , Análise Multivariada , Mutação , Miocárdio/patologia , Proteínas do Tecido Nervoso/genética , FenótipoRESUMO
BACKGROUND: Fischer 344 (F344) rats are relatively resistant to hypoxia-induced right ventricular (RV) hypertrophy compared with the Wistar-Kyoto (WKY) strain. These 2 strains were used to examine the genetic basis for the differential response. METHODS AND RESULTS: Male F(2) offspring from an F344xWKY intercross were exposed to hypoxia (10% O(2)) for 3 weeks, and pulmonary artery pressure and cardiac chamber weights were measured. Genomic DNA was screened by use of polymorphic microsatellite markers across the whole genome (excluding the sex chromosomes). A quantitative trait locus (QTL) for RV weight was identified on rat chromosome 17 (lod score 6.5) that accounted for 22% of the total variance of RV weight in the F(2) population and was independent of pulmonary artery pressure. The peak was centered over marker D17Rat41, close to Chrm3, with a 1-lod support interval of 5 cM. Comparison of homologous regions in mice and humans suggested that Ryr2, the cardiac isoform of the ryanodine receptor, colocalizes with our QTL. A panel of somatic cell hybrids and fluorescence in situ hybridization mapped Ryr2 close to the gene Chrm3 within our QTL. [(3)H]Ryanodine binding to cardiac membranes from the parental strains showed a 21% reduction in B(max) in the WKY compared with the F344 strain, with no difference in K:(d). CONCLUSIONS: These data provide the first demonstration of a QTL linked to the RV response to hypoxia-induced pulmonary hypertension. The Ryr2 receptor gene lies within this QTL and merits further investigation as a candidate for this differential RV response.
Assuntos
Hipertensão Pulmonar/complicações , Hipertrofia Ventricular Direita/complicações , Canal de Liberação de Cálcio do Receptor de Rianodina/genética , Animais , Peso Corporal , Cromossomos Humanos Par 17 , Cruzamentos Genéticos , Ligação Genética , Humanos , Hipertensão Pulmonar/etiologia , Hipertensão Pulmonar/genética , Hipertrofia Ventricular Direita/genética , Hipóxia , Hibridização in Situ Fluorescente , Masculino , Miocárdio/metabolismo , Tamanho do Órgão , Fenótipo , Característica Quantitativa Herdável , Ensaio Radioligante , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos WKY , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismoRESUMO
OBJECTIVE: Methylation of cytosine in CG dinucleotides within regulatory elements is believed to silence gene expression. These dinucleotides occur in certain important regulatory elements in the promoter region of the human beta-myosin heavy chain (beta-MHC) gene. We therefore investigated whether methylation of these elements correlates with beta-MHC gene transcription in human 'expressing' (right atrial) and 'non-expressing' (peripheral blood leucocytes) cells. METHODS: We employed 2 techniques to assess promoter methylation: (i) analysis of the susceptibility to digestion of a particular CCGG restriction site in the promoter region when genomic DNA is cleaved with the restriction endonucleases MspI (methylation-insensitive) and HpaII (methylation-sensitive), and (ii) the bisulphite-PCR method to examine in detail the methylation patterns of 3 important regulatory elements that contain CG dinucleotides. beta-MHC mRNA expression in right atrium and leucocytes was assessed using reverse-transcription-PCR with specific primers that do not detect alpha-MHC cDNA. RESULTS: The digestion pattern observed with MspI or HpaII indicated that the CCGG site was almost completely methylated in leucocytes, but relatively unmethylated in atrial myocardium from the same patients. When methylation was examined with the bisulphite-PCR method we found a reciprocal relationship between the level of beta-MHC mRNA expression in leucocytes and atrial myocardium and the degree of methylation of CG dinucleotides in the 5' regulatory elements of the gene. CONCLUSIONS: Tissue-specific methylation of the human beta-MHC gene promoter may play a role in determining the pattern of expression of this gene. Furthermore, alteration of the level of methylation may underlie the changes in transcription of this gene that occur, for example, when atrial or ventricular myocardium hypertrophies.
Assuntos
Cardiomiopatia Hipertrófica/metabolismo , Metilação de DNA , Cadeias Pesadas de Miosina/genética , RNA Mensageiro/metabolismo , Sequências Reguladoras de Ácido Nucleico , Transcrição Gênica , Cardiomiopatia Hipertrófica/genética , Expressão Gênica , Humanos , Leucócitos/metabolismo , Miocárdio/metabolismo , Reação em Cadeia da PolimeraseRESUMO
OBJECTIVE: At present little is known about the factors that regulate the expression of the endothelins and their receptors in cardiac tissue in vivo. The aim of this study was to investigate changes in expression of the endothelins (ET-1, ET-2, and ET-3) and their receptors (ETAR and ETBR) in the hypertrophied heart of the aortovenocaval (AV) fistula rat. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to quantify cardiac mRNA expression of the endothelins and their receptors during the development of cardiac hypertrophy, while radioligand binding was employed to quantify the amount of [125I]-ET-1 binding to cardiac membranes. Tissue and plasma concentrations of ET-1 were measured by radioimmunoassay. RESULTS: In control sham operated animals, ET-1 mRNA was approximately fivefold greater in atria than in ventricles (P < 0.05), but there were no atrioventricular differences in ET-2 or ET-3 mRNA. In the AV fistula rats there was a prompt three- to fourfold increase in ET-1 mRNA in atria and a progressive five- to sevenfold rise in ventricles during cardiac hypertrophy. There were no changes in ET-2 or ET-3 transcript prevalences, except for a late rise (35 d) in ET-2 mRNA levels in left ventricle. Consistent with ET-1 mRNA measurements, immunoreactive endothelin levels were increased by 7 d in atria, but not in ventricles. In control rat hearts, ETAR mRNA levels were similar in atria and ventricles, but the prevalence of ETBR was approximately sevenfold greater in the former. ETAR mRNA prevalence increased with hypertrophy in all chambers, while ETBR transcript levels were raised only in the right ventricle. There was no significant difference in [125I]-ET-1 binding between atrial samples from 35 d control and 35 d AV fistula rats, suggesting rapid turnover of endothelin receptors balanced by increased transcription from the ETAR gene. CONCLUSIONS: During cardiac hypertrophy in AV fistula rats there is increased activity of the endothelin system mediated principally by ET-1 and the ETAR subtype.
Assuntos
Cardiomegalia/metabolismo , Endotelinas/metabolismo , Miocárdio/metabolismo , Receptores de Endotelina/metabolismo , Animais , Sequência de Bases , Primers do DNA/genética , Modelos Animais de Doenças , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Ratos , Ratos Wistar , Receptor de Endotelina ARESUMO
OBJECTIVES: This study examined the expression of the C-type receptor for the natriuretic peptide family (NPR-C) in the ventricles of normal and aortovenocaval (AV)-fistula rats, the latter a model of cardiac volume overload producing hypertrophy of both ventricles. METHODS: Western blotting with a rabbit anti-NPR-C antibody was used to quantify NPR-C levels in ventricular membranes. NPR-C expression was localised anatomically and measured in frozen sections of cardiac tissue by histochemistry and in vitro autoradiography. RESULTS: Western blot analysis revealed a single band (approximately 120 kDa) in ventricular membranes which was reduced to approximately 60 kDa after treatment with beta-mercaptoethanol. NPR-C immunoreactivity and [125I]rat ANP1-28 binding (displaceable by the NPR-C-specific ligand C-ANP 4-23) were localised to the endocardium. NPR-C protein levels, as measured by all three techniques, were reduced significantly in the hypertrophied ventricles of AV-fistula rats compared to sham-operated animals. CONCLUSIONS: Volume-induced cardiac hypertrophy in the AV-fistula rat is associated with downregulation of endocardial NPR-C. This may be one mechanism by which the endocardium regulates the myocardial response to changes in haemodynamic load.
Assuntos
Fístula Arteriovenosa/metabolismo , Cardiomegalia/metabolismo , Guanilato Ciclase/metabolismo , Miocárdio/metabolismo , Receptores do Fator Natriurético Atrial/metabolismo , Animais , Fístula Arteriovenosa/complicações , Autorradiografia , Western Blotting , Cardiomegalia/etiologia , Regulação para Baixo , Imuno-Histoquímica , Masculino , Ligação Proteica , Ratos , Ratos WistarRESUMO
Genetic determinants affect adult cardiac mass and the predisposition to develop cardiac hypertrophy. The aim of this study was to identify quantitative trait loci (QTL) that control heart and left ventricular (LV) weight by use of normotensive inbred rat strains that differ in their adult cardiac mass phenotype. We studied 126 male F2 rats derived from a cross of normotensive Wistar-Kyoto and Fischer 344 rats. At 12 weeks of age, total heart weight and LV weight were measured. Genomic DNA from these animals was screened by use of polymorphic microsatellite markers across the whole genome (excluding the sex chromosomes). In this cross, the genetic contribution to total heart weight variation was 56%, and the genetic contribution for LV weight was 55%. Using the Mapmaker/QTL computer package, we identified a significant QTL on chromosome 3 with a log10 likelihood (LOD) score of 4.8, which accounted for 16.5% of the total variance of LV weight. This QTL was centered close to the marker D3Rat29. The QTL was also found to be significantly linked with total heart weight (LOD=4.4). These data provide the first demonstration of a QTL on chromosome 3 that plays a role in determining the difference in LV mass between normotensive Fischer 344 and Wistar- Kyoto inbred rat strains. The prostaglandin synthase 1 gene is located within the QTL.
Assuntos
Cardiomegalia/genética , Animais , Mapeamento Cromossômico , Cruzamentos Genéticos , Feminino , Ligação Genética , Masculino , Fenótipo , Ratos , Ratos Endogâmicos Dahl , Ratos Endogâmicos F344 , Ratos Endogâmicos WKYRESUMO
1. Angiotensin II (AII) binding density and the effect of chronic AII receptor blockade were examined in the rat model of hypoxia-induced pulmonary hypertension. 2. [125I]-[Sar1,Ile2]AII binding capacity was increased in lung membranes from rats exposed to hypoxia (10% fractional inspired O2) for 7 days compared to normal rats (Bmax 108 +/- 12 vs 77 +/- 3 fmol mg-1 protein; P < 0.05), with no significant change in dissociation constant. Competition with specific AII receptor subtype antagonists demonstrated that AT1 is the predominant subtype in both normal and hypoxic lung. 3. Rats treated intravenously with the AT1 antagonist, GR138950C, 1 mg kg-1 day-1 rather than saline alone during 7 days of exposure to hypoxia developed less pulmonary hypertension (pulmonary arterial pressure: 21.3 +/- 1.7 vs 28.3 +/- 1.1 mmHg; P < 0.05), right ventricular hypertrophy (right/left ventricle weight ratio: 0.35 +/- 0.01 vs 0.45 +/- 0.01; P < 0.05) and pulmonary artery remodelling (abundance of thick-walled pulmonary vessels: 9.6 +/- 1.4% vs 20.1 +/- 0.9%; P < 0.05). 4. The reduction in cardiac hypertrophy and pulmonary remodelling with the AT1 antagonist was greater than that achieved by a dose of sodium nitroprusside (SNP) that produced a comparable attenuation of the rise in pulmonary arterial pressure during hypoxia. 5. The data suggest that AII, via the AT1 receptor, has a role in the early pathogenesis of hypoxia-induced pulmonary hypertension in the rat.
Assuntos
Angiotensina II/fisiologia , Hipertensão Pulmonar/etiologia , Hipóxia/metabolismo , Pulmão/química , Receptores de Angiotensina/análise , Ácido 15-Hidroxi-11 alfa,9 alfa-(epoximetano)prosta-5,13-dienoico , Antagonistas de Receptores de Angiotensina , Animais , Benzofuranos/farmacologia , Masculino , Nitroprussiato/farmacologia , Endoperóxidos Sintéticos de Prostaglandinas/farmacologia , Ratos , Ratos Wistar , Receptores de Angiotensina/fisiologia , Tromboxano A2/análogos & derivados , Tromboxano A2/farmacologiaRESUMO
Conscious rats were given an intraperitoneal (i.p.) injection of isotonic or hypertonic saline, and killed 10-240 min later. In the hypothalamus of hypertonic saline-injected rats, c-fos-mRNA positive cells were mainly restricted to the supraoptic and paraventricular nuclei and to structures associated with the lamina terminalis of the third ventricle, including in particular the subfornical organ, the organum vasculosum of the lamina terminalis (OVLT) and the median preoptic nucleus. These structures comprise the proposed anterior hypothalamic 'osmoreceptor complex' for regulation of vasopressin release. The time course of the appearance and disappearance of c-fos mRNA signals was similar in all regions. Thus c-fos protein (Fos) may be a common transcription factor in the hypothalamic neural circuits involved in osmoregulation.
Assuntos
Genes fos/genética , Hipotálamo/fisiologia , RNA Mensageiro/biossíntese , Fatores de Transcrição/genética , Equilíbrio Hidroeletrolítico/fisiologia , Animais , Ventrículos Cerebrais/metabolismo , Masculino , Vias Neurais/fisiologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Ratos , Ratos Endogâmicos , Solução Salina Hipertônica , Núcleo Supraóptico/metabolismoRESUMO
OBJECTIVE: To document the cardiac phenotype associated with Friedreich's ataxia, a recessively inherited disorder characterised by spinocerebellar degeneration. SETTING: Individuals with Friedreich's ataxia who accepted the invitation to participate in the study. HYPOTHESIS: The cardiomyopathy associated with Friedreich's ataxia may offer a human model for the study of factors modulating cardiac hypertrophy. METHODS: 55 patients (mean (SD) age 30 (9) years) with a clinical diagnosis of Friedreich's ataxia were studied by clinical examination, electrocardiography, cross sectional and Doppler echocardiography, and analysis of the GAA repeat in the first intron of the frataxin gene. RESULTS: A wide variety of cardiac morphology was documented. Subjects with normal frataxin alleles had no evidence of cardiomyopathy. In homozygous subjects, a relation was found between the thickness of the interventricular septum (r = 0.53, p < 0.005), left ventricular mass (r = 0.48, p < 0.01), and the number of GAA repeats on the smaller allele of the frataxin gene. No relation was shown between the presence of electrocardiographic abnormalities (mainly repolarisation changes) and either the pattern of ventricular hypertrophy (if present) and degree of neurological disability or the length of time since diagnosis. No tendency to ventricular thinning or dilatation with age was found. Although ventricular systolic function appeared impaired in some cases, Doppler studies of ventricular filling were within the normal range for age. CONCLUSIONS: The cardiomyopathy associated with Friedreich's ataxia shows a variable phenotype which is not concordant with the presence of ECG abnormalities or the neurological features of the condition. As the genetic basis for Friedreich's ataxia has been established, further studies will help to clarify the molecular mechanisms of the cardiac hypertrophy.
Assuntos
Cardiomegalia/etiologia , Ataxia de Friedreich/complicações , Proteínas de Ligação ao Ferro , Adolescente , Adulto , Cardiomegalia/genética , Cardiomegalia/patologia , Ecocardiografia Doppler , Eletrocardiografia , Feminino , Ataxia de Friedreich/genética , Ataxia de Friedreich/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Miocárdio/patologia , Fenótipo , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Estudos Prospectivos , Repetições de Trinucleotídeos , FrataxinaRESUMO
The focus of this study was to examine the functional role of the unusual peripheral substitution of heme A. The effects of heme A stereochemistry on the reconstitution of the porphyrin have been examined in the heme A-apo-myoglobin complex using optical absorption and resonance Raman and electron paramagnetic resonance spectroscopies. The addition of one equivalent of heme A to apo-Mb produces a complex which displays spectroscopic signals consistent with a distribution of high- and low-spin heme chromophores. These results indicate that the incorporation of heme A into apo-Mb significantly perturbs the protein refolding.
Assuntos
Heme/análogos & derivados , Mioglobina/química , Análise Espectral , Animais , Bovinos , Dimetil Sulfóxido , Espectroscopia de Ressonância de Spin Eletrônica , Compostos Férricos/química , Compostos Ferrosos/química , Heme/química , Conformação Proteica , Análise Espectral Raman , Relação Estrutura-AtividadeRESUMO
Circulating atrial natriuretic factor (ANF) may interact with binding sites in many tissues to produce the characteristic pharmacological effects seen after intravenous administration. In addition to hormonal effects, ANF may have a local regulatory role. We have examined this hypothesis by using techniques designed to localise specific binding sites (defining target cells) and the messenger ribonucleic acid (mRNA) for ANF (defining cells with the capacity to synthesise ANF). We have demonstrated a topological relationship between the distribution of binding sites and mRNA transcripts in the heart and adrenal gland. This data supports functional evidence that ANF may have local effects, possibly including the regulation of both cardiac rhythmicity and contractility and its own secretion from the heart, and the maintenance of the cortical zones of adrenal steroid production.
Assuntos
Fator Natriurético Atrial/fisiologia , Glândulas Suprarrenais/metabolismo , Animais , Fator Natriurético Atrial/genética , Fator Natriurético Atrial/metabolismo , Coração/fisiologia , Humanos , Biologia Molecular , Miocárdio/citologia , Miocárdio/metabolismo , RNA Mensageiro/análise , Receptores do Fator Natriurético Atrial , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismoRESUMO
GSK2374697 is a genetically engineered fusion protein of a human domain antibody to exendin-4. This molecule binds with a high affinity to human serum albumin, creating a long-duration glucagon-like peptide-1 (GLP-1) receptor agonist. This study is the first evaluation of the albumin-binding domain antibody (AlbudAb) drug delivery platform in humans. The aim of this randomized clinical study was to determine the pharmacokinetics, pharmacodynamics, safety, and tolerability of GSK2374697. The pharmacokinetic profile was prolonged, with estimated half-lives ranging from 6 to 10 days. Postprandial glucose and insulin were reduced, and gastric emptying was delayed in healthy subjects, confirming anticipated GLP-1 receptor agonist pharmacology. The safety and tolerability were as expected for a potent GLP-1 agonist. Gradual titration of doses greatly improved tolerability. Rapid tolerance to nausea was observed. Study results support further investigation in type 2 diabetes and for weight loss.