RESUMO
OBJECTIVES: Immunoglobulin (Ig)G4-related disease is a recently proposed systemic disorder that includes autoimmune pancreatitis (AIP), Mikulicz's disease, and various other organ lesions. In the present retrospective study, we examined whether thyroid lesions should also be included in IgG4-related disease (Ig4-RD) under the new term IgG4-related thyroiditis. METHOD: We enrolled 114 patients with Ig4-RD, including 92 patients with AIP, 15 patients with Mikulicz's disease, and seven patients with IgG4-related cholangitis, and analysed clinical findings, function, serum values of activity markers, computed tomography (CT) images, and histology of the thyroid gland. RESULTS: Among the 22 patients (19%) in our cohort who were found to have hypothyroidism [thyroid stimulating hormone (TSH) > 4 mIU/L], 11 patients had clinical hypothyroidism [free thyroxine (FT4) < 1 ng/dL] and 11 patients had subclinical hypothyroidism (FT4 ≥ 1 ng/dL). Serum concentrations of IgG, IgG4, circulating immune complex (CIC), and ß2-microglobulin (ß2-MG) were significantly higher in the hypothyroidism group compared with the remaining 92 euthyroid patients, and serum C3 concentration was significantly lower. After prednisolone treatment, TSH values had decreased significantly (p = 0.005) in this group and FT4 values had increased significantly (p = 0.047). CT images showed that the thyroid glands of patients with clinical hypothyroidism had a significantly greater volume than those of the euthyroid and other groups. Pathological analysis of one resected thyroid gland disclosed a focused lesion with infiltration of lymphocytes and IgG4-bearing plasma cells and loss of thyroid follicles. CONCLUSIONS: Thyroid lesions associated with hypothyroidism can be considered as a new disease termed IgG4-related thyroiditis. Awareness of this condition should lead to appropriate corticosteroid treatment that may prevent progression to a fibrous state.
Assuntos
Doenças Autoimunes/diagnóstico , Hipotireoidismo/diagnóstico , Hipotireoidismo/imunologia , Imunoglobulina G/imunologia , Doença de Mikulicz/diagnóstico , Pancreatite/diagnóstico , Tireoidite Autoimune/diagnóstico , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Doenças Autoimunes/complicações , Colangite/complicações , Colangite/diagnóstico , Colangite/imunologia , Estudos de Coortes , Progressão da Doença , Feminino , Humanos , Hipotireoidismo/complicações , Masculino , Pessoa de Meia-Idade , Doença de Mikulicz/complicações , Doença de Mikulicz/imunologia , Pancreatite/complicações , Pancreatite/imunologia , Prognóstico , Estudos Retrospectivos , Medição de Risco , Fatores Sexuais , Estatísticas não Paramétricas , Testes de Função Tireóidea , Tireoidite Autoimune/complicações , Tireoidite Autoimune/imunologiaRESUMO
alpha-Adrenergic agonists such as norepinephrine and clonidine, but not methoxamine, inhibited the increase in cAMP accumulation elicited by TSH in porcine thyroid slices. The inhibitory effect of norepinephrine was abolished by yohimbine but not by prazosin. Likewise, the in vitro release of T4 from the pig thyroid slice stimulated by TSH was inhibited by norepinephrine and the inhibition was antagonized by yohimbine but not by prazosin. These results suggest that the inhibitory effect of alpha-adrenergic agonists is mediated through alpha 2-adrenoceptors. Preincubation of the thyroid slices with islet-activating protein, pertussis toxin, decreased the inhibitory effect of norepinephrine on the cAMP content, suggesting the involvement of guanine nucleotide regulatory protein (Ni) in the inhibition mediated through alpha 2-adrenoceptors. Binding studies with [3H]yohimbine and [3H]prazosin to porcine thyroid membranes revealed the specific and saturable binding of both radioligands. Scatchard analysis of the specific binding revealed a single class of binding sites for each radioligand; however, the maximum number of binding sites for [3H]yohimbine was about 10 times more than that for [3H]prazosin. In accordance with the pharmacological results, binding studies showed a predominance of alpha 2-adrenoceptors in the pig thyroid.
Assuntos
Receptores Adrenérgicos alfa/metabolismo , Glândula Tireoide/metabolismo , Animais , Toxinas Bacterianas/farmacologia , Clonidina/farmacologia , AMP Cíclico/metabolismo , Cinética , Norepinefrina/farmacologia , Toxina Pertussis , Prazosina/metabolismo , Suínos , Tireotropina/farmacologia , Tiroxina/metabolismo , Fatores de Virulência de Bordetella , Ioimbina/metabolismoRESUMO
TSH enhanced the release of T4 from the mouse thyroid incubated in vitro. Norepinephrine, a nonspecific alpha-adrenergic agonist, and methoxamine, an alpha 1-agonist, inhibited the TSH-stimulated release of T4 at 10(-4) and 10(-5) M, whereas clonidine, an alpha 2-agonist, exerted a weak inhibition. The inhibitory effect of 10(-5) M norepinephrine on the T4 release stimulated by TSH was prevented by prazosin, an alpha 1-antagonist, at concentrations higher than 10(-7) M, whereas yohimbine, an alpha 2-antagonist, exerted weak activity in antagonizing the inhibition induced by norepinephrine. Norepinephrine, methoxamine, and clonidine did not significantly reduce the cAMP accumulation stimulated by TSH in the mouse thyroid incubated in vitro. These findings in the mouse thyroid indicate that catecholamines act by way of alpha 1-adrenergic receptors to suppress TSH-stimulated release of T4 without reducing the cAMP levels stimulated by TSH in the mouse thyroid.
Assuntos
Clonidina/farmacologia , Metoxamina/farmacologia , Norepinefrina/farmacologia , Receptores Adrenérgicos alfa/fisiologia , Receptores Adrenérgicos/fisiologia , Glândula Tireoide/metabolismo , Tireotropina/farmacologia , Tiroxina/metabolismo , Animais , AMP Cíclico/metabolismo , Cinética , Masculino , Camundongos , Camundongos Endogâmicos , Prazosina/farmacologia , Receptores Adrenérgicos alfa/efeitos dos fármacos , Glândula Tireoide/efeitos dos fármacos , Ioimbina/farmacologiaRESUMO
Monolayer cultures of mouse adrenal tumor cell line Y-1 have been used to investigate the effects of glucocorticoid on cell replication, [3H]thymidine incorporation into the trichloroacetic acid-precipitated cell fraction, steroidogenesis, and the ACTH receptors of adrenocortical cells. Corticosterone at a concentration of 5.0--50 micrograms/ml inhibited cell replication and [3H]thymidine incorporation into trichloroacetic acid-precipitated cell fraction in a dose-related manner. Corticosterone at a concentration of 0.5--50 micrograms/ml inhibited ACTH-induced steroidogenesis in a dose-related manner. Steroids which do not possess glucocorticoid action did not show such inhibitory effects on cell replication and steroidogenesis of Y-1 cells. The characteristics of the ACTH receptors of these cells remained unaffected by corticosterone. Our findings suggest that synthesized or secreted glucocorticoid may play an important role in the direct regulation of proliferation and function of adrenocortical cells under physiological conditions.
Assuntos
Neoplasias das Glândulas Suprarrenais/metabolismo , Corticosterona/farmacologia , Esteroides/biossíntese , Hormônio Adrenocorticotrópico/metabolismo , Animais , Divisão Celular/efeitos dos fármacos , Linhagem Celular , DNA de Neoplasias/biossíntese , Cinética , Camundongos , Receptores de Superfície Celular/metabolismo , Timidina/metabolismoRESUMO
Hypertension is commonly associated with diabetes mellitus. The aim of the present study was to explore the pathophysiological significance of the natriuretic peptide (NP) system in hypertension associated with genetically obese/hyperglycemic Wistar fatty rats. The messenger RNA (mRNA) levels of the two biologically active NP receptors, NP-A receptor [more specific for atrial natriuretic peptide (ANP)] and NP-B receptor [more specific for C-type natriuretic peptide (CNP)], and CNP mRNA levels were determined in the aorta and kidney by ribonuclease protection assay. Plasma ANP levels were determined by RIA. Both NP-A and NP-B receptor mRNA levels in the aortae of Wistar fatty rats were double those in Wistar lean rats. Plasma ANP levels and CNP mRNA levels in the aorta of Wistar fatty rats were also significantly higher than those in Wistar lean rats. In contrast, there was no significant difference in renal levels of the mRNA for both NP receptors and CNP between the two strains. Administration of a NP-A and -B receptor antagonist, HS-142-1, to Wistar fatty rats resulted in a significant increase in systolic blood pressure and a larger decrease in plasma cGMP level than that in Wistar lean rats, with no difference in the extents of decrease in urine volume and urinary sodium excretion between the two strains. These results suggest that both the ANP/NP-A system and the CNP/NP-B system in vessels are up-regulated at the level of gene expression and may, thus, play an important role in counteracting the hypertension associated with diabetes mellitus.
Assuntos
Fator Natriurético Atrial/fisiologia , Vasos Sanguíneos/fisiopatologia , Hiperglicemia/fisiopatologia , Hipertensão/fisiopatologia , Obesidade/fisiopatologia , Proteínas/fisiologia , Animais , Aorta/metabolismo , Fator Natriurético Atrial/sangue , GMP Cíclico/sangue , Diurese/efeitos dos fármacos , Guanilato Ciclase/genética , Guanilato Ciclase/metabolismo , Hiperglicemia/genética , Hipertensão/induzido quimicamente , Hipertensão/genética , Rim/metabolismo , Masculino , Natriurese/efeitos dos fármacos , Peptídeo Natriurético Tipo C , Obesidade/genética , Polissacarídeos/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores do Fator Natriurético Atrial/genética , Receptores do Fator Natriurético Atrial/metabolismoRESUMO
Receptors for natriuretic peptide (NP) consist of three subtypes: NP-A, NP-B, and NP-C. Recent studies in cultured aortic cells have suggested a phenotype-related switching of the vascular NP receptor from NP-A to NP-B. To ascertain the biological significance of the phenomenon in vivo, we developed a sensitive and reproducible ribonuclease protection assay and determined each receptor messenger RNA (mRNA) level in the vascular vessels of stroke-prone spontaneously hypertensive rats, deoxycorticosterone acetate-salt hypertensive rats, and genetically hyperglycemic. Wistar fatty rats and in cultured aortic smooth muscle cells. The aortic NP-A receptor mRNA level was significantly up-regulated in both types of hypertensive rats, whereas the NP-B receptor mRNA level did not show any significant change. Both NP-A and NP-B receptor mRNA levels were significantly up-regulated in Wistar fatty rats compared with the control values. There was no significant up-regulation of NP-A receptor mRNA in the inferior vena cava of the stroke-prone spontaneously hypertensive rats. Although the NP-A receptor was always the predominant subtype in rat aortic tissue, NP-B receptor was the predominant subtype in aortic smooth muscle cells in culture. These findings suggest that up-regulation of the NP-A receptor, but not the subtype switching, is the major modulation of receptor gene expression in both hypertensive and diabetic rats.
Assuntos
Hipertensão/genética , Obesidade/genética , Receptores do Fator Natriurético Atrial/genética , Receptores de Peptídeos/genética , Animais , Aorta Torácica/metabolismo , Sequência de Bases , Células Cultivadas , Primers do DNA/genética , Expressão Gênica , Guanilato Ciclase/genética , Hiperglicemia/genética , Hiperglicemia/metabolismo , Hipertensão/metabolismo , Masculino , Dados de Sequência Molecular , Músculo Liso Vascular/metabolismo , Obesidade/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Receptores de Peptídeos/classificação , Ribonucleases , Regulação para Cima , Veia Cava Inferior/metabolismoRESUMO
Biological actions of natriuretic peptide (NP) are determined by the condition of the receptor as well as that of the hormone. Although we previously demonstrated in hypertensive rats the up-regulation of NP-A receptor that mediates various biological actions of NPs, the pathophysiologic significance of NP-C receptor, another subtype thought to be related to clearance of NPs and possibly to biological actions, remains unknown. In the present study, we determined NP-C receptor messenger RNA (mRNA) level in the aortic tissue of stroke-prone spontaneously hypertensive rats (SHR-SP/Izm) and in cultured aortic smooth muscle cells by ribonuclease protection assay. The aortic NP-C receptor mRNA level in SHR-SP/Izm was significantly lower than that in the control WKY/Izm. Oral administration of an angiotensin (Ang) II receptor (AT1) antagonist, TCV-116, but not a calcium channel blocker, manidipine, reversed the down-regulated NP-C receptor mRNA in SHR-SP/Izm to the level in WKY/Izm, whereas the latter was more potent in decreasing the blood pressure. In cultured aortic smooth muscle cells, the NP-C receptor was the predominant subtype. Ang II decreased the NP-C receptor mRNA level in a dose-dependent manner, but this effect was reversed by an AT1 antagonist, CV-11974. Neither the NP-A nor NP-B receptor mRNA level was altered by Ang II. These findings indicate that vascular NP-C receptor is down- regulated via Ang-II-mediated mechanism in SHR-SP/Izm. The phenomenon, together with the up-regulation of the NP-A receptor, may play an important role in counteracting hypertension by enhancing the action of NPs.
Assuntos
Angiotensina II/metabolismo , Guanilato Ciclase/metabolismo , Hipertensão/metabolismo , Músculo Liso Vascular/metabolismo , Proteínas/metabolismo , Receptores do Fator Natriurético Atrial/metabolismo , Tetrazóis , Angiotensina II/farmacologia , Antagonistas de Receptores de Angiotensina , Animais , Benzimidazóis/farmacologia , Compostos de Bifenilo/farmacologia , Bloqueadores dos Canais de Cálcio/farmacologia , Células Cultivadas , Di-Hidropiridinas/farmacologia , Regulação para Baixo , Hipertensão/patologia , Masculino , Músculo Liso Vascular/patologia , Peptídeo Natriurético Tipo C , Nitrobenzenos , Piperazinas , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos SHR , Ratos WistarRESUMO
Treatment with a beta-adrenergic blocker (beta-blocker) in hypertension is associated with increased plasma atrial natriuretic peptide (ANP) levels despite a decrease in cardiac overload. The mechanism and pathophysiological significance of the phenomenon remain unclear. To clarify the role of the ANP system in the antihypertensive effects of the beta-blocker, we investigated the effects of carvedilol (30 mg/kg x day, orally, for 4 weeks) on the ANP system in stroke-prone spontaneously hypertensive rats (SHR-SP/Izm). Plasma ANP levels showed a significant increase despite a significant decrease in blood pressure and heart rate in the carvedilol group. Although ANP messenger RNA levels in the heart did not change, messenger RNA levels of the natriuretic peptide-C (NP-C) receptor as a clearance receptor showed a significant decrease in both the aorta and lung in the carvedilol group. NP-C receptor densities were also significantly decreased in the lung in this group. The biological half-life of exogenous ANP in circulating blood was prolonged in the carvedilol group compared with that in the control group. Administration of the ANP receptor antagonist, HS-142-1, resulted in a greater increase in systolic blood pressure in the carvedilol group than in the control group. In addition, both basal and ANP-stimulated cGMP contents in the aorta were significantly higher in the carvedilol group. These results suggest that carvedilol potentiates the hypotensive action of ANP by increasing plasma ANP levels and enhancing the vascular response to ANP. These effects were closely related to the down-regulation of the NP-C receptor. The newly found mechanism seems to account for a sizable portion of the antihypertensive effects of carvedilol and could be of potential importance in the treatment of cardiovascular disease with beta-blockers.
Assuntos
Antagonistas Adrenérgicos beta/farmacologia , Anti-Hipertensivos/farmacologia , Fator Natriurético Atrial/sangue , Carbazóis/farmacologia , Hipertensão/tratamento farmacológico , Propanolaminas/farmacologia , Animais , Carvedilol , GMP Cíclico/biossíntese , Masculino , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos SHR , Receptores do Fator Natriurético Atrial/genéticaRESUMO
The nature of the TSH receptor in adenoma and carcinoma of the thyroid gland was studied using a radioreceptor assay technique. A membrane fraction of tissue homogenate was obtained by discontinuous sucrose gradient ultracentrifugation, and 125I-TSH, labelled by a lactoperoxidase method, was purified with a receptor adsorption method. Both the capacities and the association constants of high affinity receptors (4 x 10(9) M-1) and of low affinity receptors (0.073 x 10(9) M-1) observed in the normal thyroid were almost identical to those of the thyroid of Graves' disease and those of thyroid adenoma. Although the two papillary carcinomas examined were found to have two kinds of TSH receptors, one of the carcinomas showed decreased association constants for both high affinity and low affinity receptors.
Assuntos
Receptores de Superfície Celular , Neoplasias da Glândula Tireoide/metabolismo , Tireotropina/metabolismo , Adenoma/metabolismo , Carcinoma Papilar/metabolismo , Doença de Graves/metabolismo , HumanosRESUMO
TSH receptor and adenylate cyclase activity of plasma membrane fractions of human thyroid tumors were examined. The mean (+/- SD) basal adenylate cyclase activity in normal thyroid tissues was 0.35 +/- 0.33 nmole/mg protein x 10 min. The activity rose to 280% (range, 270-310%) of basal with TSH (166 mU/ml). In adenomas, the activity rose to 600% (range, 530-650), which was significantly higher than that of normal thyroid (P < 0.005). In the differentiated carcinoma, TSH responsiveness of adenylate cyclase was heterogenous (range, 110-520), but was qualitatively similar to that of the normal thyroid. On the other hand, basal adenylate cyclase activity of undifferentiated carcinoma was significantly lower than that of normal thyroid (0.018 +/- 0.007 nmol/mg protein x 10 min; P < 0.05) and was not stimulated by TSH. LH, FSH, and ACTH did not stimulate the enzyme in either kind of carcinomas. The mean (+/- SD) of the capacity of the high affinity receptor of adenomas (0.72 +/- 0.64 pmol/mg protein) and differentiated carcinomas (0.77 +/- 0.84) was not significantly different from that of normal thyroid (0.92 +/- 0.84). The affinity constants of the receptors in these three tissues were much the same (1.6-2.4 x 10(10) M-1). On the other hand, high affinity receptor could not be detected in all of the undifferentiated thyroid carcinoma. It seems likely that the failure of adenylate cyclase to respond to TSH in undifferentiated carcinoma of the thyroid is due to an alteration at the level of the receptor site. These data suggest that growth and metabolic activity of undifferentiated carcinoma may be independent of TSH, while those of adenoma and differentiated carcinoma may be affected by TSH.
Assuntos
Adenilil Ciclases/metabolismo , Carcinoma/metabolismo , Receptores de Superfície Celular/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Tireotropina/farmacologia , Adenoma/metabolismo , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Humanos , Receptores da Tireotropina , Glândula Tireoide/metabolismoRESUMO
A novel insulin sensitizing agent, thiazolidine, has been demonstrated to inhibit the growth of cultured vascular smooth muscle cells (VSMC) in vitro. This study was undertaken to examine the in vivo effects of the thiazolidine compound pioglitazone (PIO) on carotid neointimal thickening, after endothelial injury in Wistar rats and vascular hypertrophy in stroke-prone spontaneously hypertensive rats (SHR-SP/Izm). PIO treatment (3 mg/kg/day for 1 week prior to endothelial injury and 2 weeks postendothelial injury) remarkably decreased neointimal cross-sectional areas in treated animals (63.8 +/- 4.9 x 10(3) microm2) versus controls (196 +/- 7.6 x 10(3) microm2, P < 0.05). Bromodeoxyuridine uptake in the neointima, a marker of DNA synthesis, was also decreased after treatment compared with controls. In SHR-SP/Izm but not in Wistar rats, PIO treatment decreased blood pressure and plasma insulin levels. PIO treatment in SHR-SP/Izm (3 mg/kg/day from 4 weeks of age for 7 weeks) significantly decreased the medial wall thickness of the mesenteric artery (10.4 +/- 1.2 x 10(3) microm2 versus control, 21.2 +/- 2.4 x 10(3) microm2, P < 0.05). In addition, PIO treatment significantly decreased the expression of EIIIA fibronectin both in the carotid neointima of Wistar rats and the media of the mesenteric artery in SHR-SP/Izm compared with their respective controls (P < 0.05). These results suggest that PIO has vasculo-protective effects in both acute and chronic vascular injury in vivo through inhibition of VSMC proliferation.
Assuntos
Arteriosclerose/prevenção & controle , Artérias Carótidas/efeitos dos fármacos , Hipertensão/prevenção & controle , Hipoglicemiantes/uso terapêutico , Tiazóis/uso terapêutico , Tiazolidinedionas , Túnica Íntima/efeitos dos fármacos , Animais , Aorta Abdominal/efeitos dos fármacos , Aorta Abdominal/metabolismo , Aorta Abdominal/patologia , Arteriosclerose/metabolismo , Arteriosclerose/patologia , Bromodesoxiuridina/metabolismo , Artérias Carótidas/metabolismo , Artérias Carótidas/patologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , DNA/biossíntese , Fibronectinas/antagonistas & inibidores , Fibronectinas/metabolismo , Hipertensão/metabolismo , Hipertensão/patologia , Hipertrofia/patologia , Hipertrofia/prevenção & controle , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/metabolismo , Artérias Mesentéricas/patologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Pioglitazona , Distribuição Aleatória , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Túnica Íntima/metabolismo , Túnica Íntima/patologiaRESUMO
To investigate the possible neuromodulatory role of nitric oxide (NO) in the gastrointestinal tract, an examination was made of the effects of NG-nitro-L-arginine (L-NOARG), an inhibitor of NO synthase, on the intestinal response to [Met5]-enkephalin (ENK) by recording the mechanical activity of the isolated duodenum from rats. [Met5]-enkephalin elicited a biphasic response of the duodenum, i.e. transient relaxation followed by contraction. The relaxation induced by ENK was blocked by naloxone, an opioid receptor antagonist, but not by tetrodotoxin (TTX). The contractile response of the duodenum to ENK was blocked by TTX but not by naloxone. The contractile response was not affected by hyoscine, a muscarinic antagonist, or guanethidine, an adrenergic neuron blocking agent, indicating mediation by non-adrenergic, non-cholinergic (NANC) nerves. The contractile but not the relaxant response to ENK was blocked by L- but not D-NOARG. The contractile response was also inhibited by methylene blue, an inhibitor of both NO synthase and guanylate cyclase, and by indomethacin, a cyclooxygenase inhibitor. Thus, endogenous NO and prostaglandins are involved in the contractile response to ENK. Endogenous NO may modulate the release of excitatory NANC transmitters via a prejunctional mechanism.
Assuntos
Sistema Nervoso Autônomo/fisiologia , Encefalina Metionina/farmacologia , Músculo Liso/fisiologia , Óxido Nítrico/fisiologia , Aminoácido Oxirredutases/antagonistas & inibidores , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Duodeno/efeitos dos fármacos , Duodeno/inervação , Duodeno/fisiologia , Encefalina Metionina/antagonistas & inibidores , Técnicas In Vitro , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/inervação , Naloxona/farmacologia , Óxido Nítrico Sintase , Nitroarginina , Ratos , Ratos Wistar , Tetrodotoxina/farmacologiaRESUMO
BACKGROUND: The immunosuppressants cyclosporine and FK506 have been used successfully in clinical transplantation, but both agents have various side effects. We have previously found that cyclosporine is prothrombotic and that FK506 is antithrombotic in an in vivo system. The aim of the present study was to assess the effects of these agents on platelet reactivity and coagulation using an in vitro shear-induced hemostatic platelet plug-forming instrument, the hemostatometer. METHODS: A purpose-built hemostatometer was constructed in our laboratory. The effects of cyclosporine and FK506 on platelet reactivity and coagulation were assessed under high shear stress using non-anticoagulated rat and human blood. RESULTS: FK506 significantly inhibited both platelet reactivity and coagulation. Cyclosporine also significantly inhibited coagulation, but a proaggregatory effect was observed at a final blood concentration of 0.05 mg/ml. CONCLUSIONS: The present in vitro results support our previous in vivo findings regarding the prothrombotic and antithrombotic effects of cyclosporine and FK506, respectively.
Assuntos
Plaquetas/fisiologia , Ciclosporina/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Tacrolimo/farmacologia , Adulto , Animais , Anticoagulantes , Coagulação Sanguínea/efeitos dos fármacos , Coagulação Sanguínea/imunologia , Plaquetas/efeitos dos fármacos , Humanos , Imunossupressores/farmacologia , Técnicas In Vitro , Masculino , Agregação Plaquetária/imunologia , Agregação Plaquetária/fisiologia , Ratos , Ratos Wistar , Estresse MecânicoRESUMO
1. Subcutaneous injection of lipopolysaccharide (LPS) increases plasma leakage in mouse skin. Pretreatment with LPS conditions mice tolerant to the LPS-induced plasma leakage. Nitric oxide (NO) has been suggested to be involved in these LPS effects. A specific role of inducible NO synthase (iNOS) was investigated in the LPS-induced plasma leakage using iNOS deficient mice. 2. Plasma leakage in mouse skin was measured by the local accumulation of Pontamine sky blue at the site of subcutaneous injection of LPS (Sal. typhimurium). LPS (100 - 400 microg site(-1)) produced a dose-related increase in dye leakage in both iNOS deficient and wild-type mice with about 40% less dye leakage in iNOS deficient mice. 3. Indomethacin (5 mg kg(-1)), N-[-2-cyclohexyloxy]-4-nitrophenyl methanesulphonamide (NS-398) (1 mg kg(-1)), diphenhydramine (10 mg kg(-1)) and anti-TNF-alpha antibody (dilution 1 : 400, 10 ml kg(-1)) inhibited the LPS-induced dye leakage in both iNOS deficient and wild-type mice, whereas N(G)-nitro-L-arginine methyl ester (L-NAME) (10 mg kg(-1)) or aminoguanidine (10 mg kg(-1)) inhibited that in wild-type but not in iNOS deficient mice. 4. Pretreatment with LPS (0.15 mg kg(-1) i.p.) 4 h before decreased the LPS-induced dye leakage in wild-type but not in iNOS deficient mice. LPS pretreatment increased serum corticosterone levels in both mice, while it increased the serum nitrate/nitrite levels in wild-type but not in iNOS deficient mice. 5. These studies indicate that an increase in vascular permeability induced by LPS is mediated by NO produced by iNOS, eicosanoids, histamine and TNF-alpha. The tolerance against LPS-induced vascular permeability change may be mediated by iNOS induction but not by an increased release of endogenous corticosteroids.
Assuntos
Permeabilidade Capilar/fisiologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico/metabolismo , Animais , Permeabilidade Capilar/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/farmacologia , Lipopolissacarídeos/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/deficiência , Óxido Nítrico Sintase Tipo II , Pele/irrigação sanguíneaRESUMO
1. Endotoxin shock is accompanied by an increase in peripheral vascular permeability. It has been postulated that most biological activities of LPS are derived from lipid A moiety. Here we examined the effect of lipid A analogue ONO-4007 in increasing vascular permeability and the possible mediators in mouse skin by a dye leakage method. 2. Subcutaneous injection of ONO-4007 (1 - 2 mg site(-1)) induced a dose-dependent increase in vascular permeability which was evident after 120 min. 3. ONO-4007-induced dye leakage was significantly attenuated by pretreatments with anti-tumour necrosis factor-alpha (TNF-alpha) and anti-interleukin-1alpha (IL-1alpha) antibodies, but not with indomethacin (5 mg kg(-1)) or diphenhydramine (10 mg kg(-1)). ONO-4007-induced dye leakage was significantly inhibited by a pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME) (10 mg kg(-1)) but not with aminoguanidine (50 mg kg(-1)). In inducible nitric oxide synthase (iNOS)-deficient mice, ONO-4007 significantly increased the dye leakage, while ONO-4007 dilated rat thoracic aortic rings pre-contracted with phenylephrine, and the L-NAME pretreatment inhibited the dilation. 4. Thus, TNF-alpha, IL-1alpha and constitutive NOSs-derived nitric oxide but not prostaglandins or histamine play a role in ONO-4007-induced increase in vascular permeability. Although ONO-4007 mimics LPS in increasing vascular permeability, mechanisms of permeability change elicited by ONO-4007 were not identical to those of LPS.
Assuntos
Permeabilidade Capilar/efeitos dos fármacos , Mediadores da Inflamação/fisiologia , Lipídeo A/análogos & derivados , Pele/efeitos dos fármacos , Animais , Anticorpos Monoclonais/farmacologia , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/fisiologia , Compostos Azo/farmacocinética , Relação Dose-Resposta a Droga , Feminino , Técnicas In Vitro , Mediadores da Inflamação/imunologia , Interleucina-1/imunologia , Interleucina-1/fisiologia , Lipídeo A/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Óxido Nítrico Sintase/deficiência , Óxido Nítrico Sintase Tipo II , Ratos , Pele/irrigação sanguínea , Fatores de Tempo , Azul Tripano , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/fisiologia , Vasodilatação/efeitos dos fármacosRESUMO
Anti-inflammatory effects of cyclic AMP elevating agents were examined in a mouse model of lipopolysaccharide (LPS)-induced microvascular permeability change. Vascular permeability on the back skin was measured by the local accumulation of Pontamine sky blue (PSB) after subcutaneous injection of LPS (400 microg site-1) from Salmonella typhimurium. Dye leakage in the skin was significantly increased 2 h after injection of LPS. This LPS-induced dye leakage was suppressed by phosphodiesterase inhibitors, including pentoxifylline (160 mg kg-1), milrinone (5 - 10 mg kg-1), rolipram (0.5 - 10 mg kg-1) and zaprinast (5 - 10 mg kg-1). The dye leakage was also inhibited by beta-adrenoceptor agonists, including isoproterenol (0.5 - 5 mg kg-1) and salbutamol (0.05 - 5 mg kg-1), an adenylate cyclase activator, forskolin (5 mg kg-1), and a cell permeable cyclic AMP analogue, 8-bromo-cyclic AMP (8-Br-cAMP, 10 mg kg-1). LPS caused a transient increase in serum TNF-alpha level peaking at 1 h after the injection. This increase in serum TNF-alpha was completely blocked by a pretreatment with pentoxifylline (160 mg kg-1), milrinone (5 mg kg-1), rolipram (1 mg kg-1), zaprinast (10 mg kg-1), salbutamol (0.5 mg kg-1), forskolin (1 mg kg-1) and 8-Br-cAMP (10 mg kg-1). LPS caused an increase in serum IL-1alpha level peaking at 3 h after injection. This increase in serum IL-1alpha was not significantly suppressed by the cyclic AMP elevating agents. Our study suggests that cyclic AMP elevating agents attenuate LPS-induced microvascular permeability change by suppressing TNF-alpha up regulation.
Assuntos
Permeabilidade Capilar/efeitos dos fármacos , AMP Cíclico/agonistas , Lipopolissacarídeos/farmacologia , Pele/metabolismo , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Adenilil Ciclases/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Colforsina/farmacologia , AMP Cíclico/análogos & derivados , Ativadores de Enzimas/farmacologia , Interleucina-1/metabolismo , Masculino , Camundongos , Inibidores de Fosfodiesterase/farmacologia , Salmonella typhimurium , Pele/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
1. Carvedilol, an adrenoceptor blocker with antioxidant activity, was studied for its ability to interact with NO in a cell-free condition and in an endothelial cell line (ECV304). 2. In a cell-free system, carvedilol attenuated NO-dependent reduction of carboxy-2-phenyl-4,4, 5,5-tetramethyl-imidazoline-1-oxyl-3-oxide induced by a NO donor, 1-hydroxy-2-oxo-3-(aminopropyl)-3-isopropyl-1-triazene (NOC5), which was determined by electron paramagnetic resonance (EPR) spectrometry. The EPR study also showed that nitrosylhaemoglobin formation in rat red blood cells by the addition of NO-saturated solution was attenuated by prior incubation with 0.1 - 10 microM carvedilol. 3. NO-induced fluorescence in 4,5-diaminofluorescein-2 diacethyl (DAF-2DA)-loaded ECV304 cells was attenuated by carvedilol but not by labetalol. The IC(50) of carvedilol for NOC5 or sodium nitroprusside-induced fluorescence of DAF-2DA in ECV304 cells was 1. 0x10(-7) M, which was similar to the reported IC(50) of carvedilol for the antioxidant effect. 4. Cell toxicity induced by a NO donor determined by the number of viable cells after 24 h treatment with 2-2'(hydroxynitrosohydrazino)bis-ethanamine was significantly attenuated by pretreatment with 1 microM carvedilol. 5. Both free and cell-associated carvedilol quenched NO. Because NO mediates both physiological and pathophysiological processes, NO quenching by the drug may have diverse clinical implications depending upon specific functions of local NO in tissues where carvedilol is distributed.
Assuntos
Carbazóis/farmacologia , Endotélio Vascular/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Óxido Nítrico/antagonistas & inibidores , Propanolaminas/farmacologia , Animais , Carvedilol , Contagem de Células/efeitos dos fármacos , Linhagem Celular , Sistema Livre de Células , Óxidos N-Cíclicos/metabolismo , Relação Dose-Resposta a Droga , Espectroscopia de Ressonância de Spin Eletrônica , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Fluoresceína , Fluorometria , Humanos , Peróxido de Hidrogênio/farmacologia , Imidazóis/metabolismo , Microscopia de Fluorescência , Óxido Nítrico/farmacologia , Óxido Nítrico/fisiologia , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , Oxirredução/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Triazenos/farmacologiaRESUMO
Serum concentrations of thyroid hormones and the properties of iodothyronine deiodinase in tissues of the house musk shrew, Suncus murinus, were examined and compared with those of the rat. Serum concentrations of thyroxine (T4) and 3,3',5'-tri-iodothyronine (rT3) were higher, while the serum concentration of 3,5,3'-tri-iodothyronine (T3) was lower in the shrew than in the rat. Among liver, kidney, skeletal muscle, heart, brown adipose tissue (BAT), spleen, lung, testes and thymus homogenates of the shrew, T(4)5'-deiodinase (5'D) activity was highest in BAT, and rT3 5'D activity was highest in both the liver and BAT. Intermediate or low T3 5-deiodinase activity was noted in all tissues examined. Activity of 5'D for T4 and rT3 in liver and kidney was much lower, while that of BAT was much higher in the shrew than in the rat. Liver and kidney 5'D may be type-I and that of BAT may be type-II in the shrew, judging from its response to 6-n-propyl-2-thiouracil and iopanoic acid and substrate preference. Thus 5'D of the shrew was similar to that of the rat in type, but was different with respect to its activity in some peripheral tissues. This difference may have relevance to the low T3 state of the shrew.
Assuntos
Iodeto Peroxidase/metabolismo , Musaranhos/metabolismo , Hormônios Tireóideos/sangue , Tecido Adiposo Marrom/metabolismo , Animais , Rim/metabolismo , Fígado/metabolismo , Masculino , Ratos , Ratos Endogâmicos , Timo/metabolismo , Tiroxina/sangue , Tri-Iodotironina/sangue , Tri-Iodotironina Reversa/sangueRESUMO
To examine the mechanism of the alpha 1-adrenoceptor-mediated inhibition of the release of thyroid hormones in the mouse thyroid, the effect of noradrenaline on the production of inositol phosphates was investigated using thyroids preloaded with [3H]inositol. Noradrenaline increased [3H]inositol accumulation into inositol phosphates in the mouse thyroid during a 30-min incubation whereas TSH (10 mU/ml) was without effect. The effect of noradrenaline was mimicked by phenylephrine, but not by clonidine or isoprenaline. Prazosin antagonized the effect of noradrenaline, while yohimbine had no effect. These results suggest that noradrenaline-induced production of inositol phosphates is mediated by alpha 1-adrenoceptors. The inhibition of release of thyroid hormones elicited by alpha 1-adrenergic agonists in the mouse thyroid is possibly mediated through the hydrolysis of membrane phosphoinositides followed by the generation of second messengers such as inositol phosphates and diacylglycerol.
Assuntos
Fosfatos de Inositol/metabolismo , Receptores Adrenérgicos alfa/fisiologia , Fosfatos Açúcares/metabolismo , Glândula Tireoide/metabolismo , Tiroxina/metabolismo , Animais , Masculino , Camundongos , Norepinefrina/farmacologia , Receptores Adrenérgicos alfa/efeitos dos fármacos , Glândula Tireoide/efeitos dos fármacosRESUMO
Morphine reduced the release of thyroid-stimulating hormone (TSH) which stimulated by exposure to cold and by thyroidectomy as well as reducing the basal level of TSH in the serum of male rats. Thi inhibitory effect of morphine was antagonized by naloxone which did not enhance the basal or cold-induced TSH release. Pretreatment with morphine did not reduce the release of TSH induced by exogenous thyrotrophin-releasing hormone (TRH) but enhanced it. This effect of morphine was also antagonized by naloxone. The above results suggested that the effect of morphine in reducing levels of serum TSH was not mediated by blocking the effect of TRH on the anterior pituitary gland, but that it was probably mediated by the inhibition of the release of TRH.