RESUMEN
Cirrhotic portal hypertension is characterized by increased hepatic oxidative stress, AA (arachidonic acid)-derived TXA(2) (thromboxane A(2)) release and exaggerated hepatic response to the α-adrenergic agonist MTX (methoxamine). Besides promoting hepatic fibrosis, the role of hyperleptinaemia in the modulation of vascular response in NASH (non-alcoholic steatohepatitis) rat livers remains unknown. The aim of the present study was to explore the possible links between hyperleptinaemia and the disarrangement in the hepatic microcirculation. NASH-cirrhosis with hyperleptinaemia was induced in lean rats by feeding with an HF/MCD (high-fat/methionine-choline-deficient) diet. Portal haemodynamics, various substances, protein and mRNA expression and PUFA (polyunsaturated fatty acid) composition were measured. Finally, the effects of leptin pre-infusion on TXA(2) release and concentration-PPP (portal perfusion pressure) curves in response to MTX were evaluated by simultaneously pre-treatment with the Kupffer cell inactivators GdCl(3) (gadolinium chloride) or EC (encapsulated clodronate), the TXS (TXA(2) synthase) inhibitor furegrelate, the TP receptor (TXA(2) receptor) antagonist SQ29548 and the dual TXS/TP receptor antagonist BM567. In HF/MCD+leptin-lean rats, cirrhosis-induced PPP and MTX hyper-responsiveness were associated with increased hepatic TXA(2) production, TBARS (thiobarbituric acid-reacting substances) levels and the AA (arachidonic acid)/n-3 PUFA ratio, and up-regulation of hepatic leptin, FAS (fatty acid synthase), NADPH oxidase subunits, TXS, TP receptor, TGFß(1) (transforming growth factor ß(1)) proteins and mRNAs. Pre-infusion of leptin significantly enhanced MTX-stimulated PPP elevation and TXA(2) release, which were attenuated by GdCl(3) and EC pre-treatment. Concomitantly pre-incubation with BM567, but not furegrelate or SQ29548, significantly abolished the leptin-enhanced MTX-stimulated increase in PPP in NASH-cirrhotic rats. Hyperleptinaemia plays an important role in hyper-responsiveness to MTX in NASH-cirrhotic rat livers with portal hypertension. The leptin-enhanced MTX-stimulated increase in PPP is mediated by increased oxidative stress and Kupffer-cell-activated AA-derived TXA(2) release in NASH-cirrhotic rats.
Asunto(s)
Hígado Graso/fisiopatología , Hipertensión Portal/fisiopatología , Macrófagos del Hígado/fisiología , Leptina/metabolismo , Metoxamina/metabolismo , Estrés Oxidativo/fisiología , Tromboxano A2/metabolismo , Análisis de Varianza , Animales , Ácido Araquidónico/metabolismo , Benzofuranos/farmacología , Colina/metabolismo , Ácido Clodrónico/farmacología , Cartilla de ADN/genética , Dieta Alta en Grasa , Ácidos Grasos Insaturados/metabolismo , Hígado Graso/complicaciones , Hígado Graso/metabolismo , Gadolinio/farmacología , Hemodinámica , Hipertensión Portal/etiología , Resistencia a la Insulina/fisiología , Macrófagos del Hígado/efectos de los fármacos , Leptina/sangre , Metionina/deficiencia , Microcirculación/fisiología , Enfermedad del Hígado Graso no Alcohólico , ARN Mensajero/metabolismo , Ratas , Receptores de Tromboxano A2 y Prostaglandina H2/antagonistas & inhibidores , Receptores de Tromboxano A2 y Prostaglandina H2/metabolismo , Compuestos de Sulfonilurea/farmacología , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
PURPOSE: The aim of this study is to evaluate the effect of retigabine on the smooth muscle response to acetylcholine, adrenaline, α-and ß-adrenoceptor agonists. METHODS: We studied the change in the spontaneous smooth muscle contraction of guinea pig gastric corpus strips before and after 20-min treatment with 2µM retigabine. We also evaluated the effect of retigabine on the smooth muscle response to 10µM acetylcholine, 1 and 10µM adrenaline, 1µM methoxamine, 0.1µM p-iodoclonidine and 10µM isoproterenol. RESULTS: We observed a significant reduction in the effects of all studied mediators and agonists when they were added to organ baths in the presence of retigabine. Retigabine diminished the effect of acetylcholine on the spontaneous smooth muscle activity. The effect was fully antagonized by XE-991 (Kv7 channel blocker), which supports our hypothesis about the role of KCNQ channels in the registered changes. The increase in the contraction force after adding of 1µM adrenaline, methoxamine, and 0.1µM p-iodoclonidine was also significantly smaller in presence of retigabine. However, comparing the effect of 10µM adrenaline on the contractility before and after treatment with retigabine, we observed increased contractility when retigabine was present in the organ baths. CONCLUSION: A possible explanation for the observed diminished effects of mediators and receptor agonists is that the effect of retigabine on smooth muscle contractility is complex. The membrane hyperpolarization, the interaction between Kv7 channels and adrenoceptors, and the influence on signaling pathways may contribute to the summary smooth muscle response.
Asunto(s)
Agonistas Adrenérgicos/farmacología , Carbamatos/farmacología , Agonistas Colinérgicos/farmacología , Músculo Liso/efectos de los fármacos , Fenilendiaminas/farmacología , Acetilcolina/metabolismo , Acetilcolina/farmacología , Animales , Antracenos/farmacología , Clonidina/análogos & derivados , Clonidina/farmacología , Interacciones Farmacológicas , Epinefrina/metabolismo , Epinefrina/farmacología , Cobayas , Isoproterenol/farmacología , Canales de Potasio KCNQ/antagonistas & inhibidores , Canales de Potasio KCNQ/metabolismo , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Metoxamina/metabolismo , Metoxamina/farmacología , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Músculo Liso/fisiología , Distribución Aleatoria , Estómago/efectos de los fármacos , Estómago/fisiología , Técnicas de Cultivo de TejidosRESUMEN
1. The alpha 1-adrenoceptor subtype mediating contraction of the rat epididymal vas deferens and rat spleen has been investigated by use of alpha 1-adrenoceptor antagonists that have shown selectivity between the different cloned receptor subtypes. 2. In the rat epididymal vas deferens the potency of noradrenaline and phenylephrine was increased in the presence of neuronal and extra-neuronal uptake blockers, cocaine and beta-oestradiol, but these did not alter that of methoxamine. The order of potency of the agonists in the presence or absence of uptake blockade was noradrenaline > phenylephrine > methoxamine. In the rat spleen the potency of these agonists was not altered in the presence of cocaine and beta-oestradiol, and their order of potency was the same as in the vas deferens. 3. The non subtype selective alpha 1-adrenoceptor antagonist prazosin (up to 1 x 10(-7) M) was found to antagonize contractions to noradrenaline in the vas deferens competitively (pA2 9.2), but only in a non competitive manner in the spleen. Contractions to phenylephrine in the spleen however were competitively antagonized by prazosin (up to 1 x 10(-7) M) with a pA2 of 9.2. This suggests that there is an alpha 1- and a non alpha 1-adrenoceptor response to noradrenaline in the rat spleen. 4. Pretreatment with chlorethylclonidine (10(-4) M for 30 min) did not alter the noradrenaline contractions in the vas deferens, but contractions to noradrenaline and phenylephrine in the spleen were shifted 30 and 300 fold to the right of the control curve, respectively. This suggests that only the contractions in the spleen were mediated by alpha 1B-adrenoceptors. 5. The noradrenaline contractions in the vas deferens were competitively antagonized by WB 4101 (pA29.6), 5-methyl-urapidil (pA2 8.7), phentolamine (pA2 8.3), benoxathian (pA2 9.4), spiperone (pA2 7.5),indoramin (pA2 8.4) and BMY 7378 (pA2 6.7), consistent with the affinities of these antagonists in binding studies on tissue alpha 1A-adrenoceptors. These values correlated best with their published affinities on the expressed alpha 1c-adrenoceptor clone and poorly with those at either the expressed alpha lb- or alpha 1d adrenoceptor clones. Therefore the classical alpha 1A-adrenoceptor appears to be the same as the expressed alpha lc-adrenoceptor clone.6. The phenylephrine contractions in the spleen were competitively antagonized by WB 4101 (pA2 8.1),5-methyl-urapidil (pA2 7.1), phentolamine (pA2 7.3), benoxathian (pA2 7.4), spiperone (pA2 7.9),indoramin (pA2 7.5) and BMY 7378 (pA2 7.4), consistent with the affinities of these antagonists in binding studies on tissue alB-adrenoceptors. The pA2 values correlated best with the published affinities of these compounds on the expressed alb-adrenoceptor clone and poorly with those at either the expressed alpha ld- or alpha lc-adrenoceptor clones. Therefore the alpha lB-adrenoceptor appears to be the same as the expressed alpha lb-adrenoceptor clone.7. The results provide pharmacological evidence that the alpha1-adrenoceptor mediating noradrenaline contractions in the epididymal portion of the rat vas deferens is the alpha 1A-(alpha lC) subtype and that contractions to phenylephrine in the rat spleen are mediated by the alpha 1B-subtype.
Asunto(s)
Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Músculo Liso/efectos de los fármacos , Receptores Adrenérgicos alfa 1/efectos de los fármacos , Bazo/efectos de los fármacos , Conducto Deferente/efectos de los fármacos , Animales , Unión Competitiva , Clonidina/análogos & derivados , Clonidina/farmacología , Cocaína/toxicidad , Epidídimo/efectos de los fármacos , Epidídimo/metabolismo , Estradiol/toxicidad , Masculino , Metoxamina/metabolismo , Metoxamina/farmacología , Contracción Muscular/efectos de los fármacos , Norepinefrina/metabolismo , Norepinefrina/farmacología , Fenilefrina/metabolismo , Fenilefrina/farmacología , Prazosina/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos alfa 1/clasificación , Receptores Adrenérgicos alfa 1/metabolismo , Bazo/metabolismo , Conducto Deferente/metabolismo , Vasoconstrictores/toxicidadRESUMEN
Ligand binding studies with WB 4101 revealed that the rat aorta contains both the alpha 1a- and alpha 1b-adrenoceptor subtypes. Results obtained following treatment with the irreversible antagonists phenoxybenzamine, chlorethylclonidine or SZL-49 (4-amino-6,7-dimethoxy-2-quinazolinyl-4-(2-bicyclo[2,2,2]octa-2,5- dienylcarbonyl-2-piperazine) suggest that there is a complex interaction between the alpha 1-adrenoceptor subtypes in the aorta. Chlorethylclonidine affects only the alpha 1b-adrenoceptor, whereas the predominant action of SZL-49 is on the alpha 1a-subtype. Chlorethylclonidine significantly inhibited the response to either methoxamine or phenylephrine, agents which are selective alpha 1a-adrenoceptor agonists. Following inactivation with either chlorethylclonidine or SZL-49, the response of the rat aorta to phenylephrine was only partially antagonized by either prazosin or WB 4101. SZL-49 also inhibited the response of the rat tail artery to electrical stimulation. The response of the tail artery obtained following inactivation with SZL-49 was effectively antagonized by prazosin. Phenylephrine, prazosin or WB 4101 afforded complete protection from chlorethylclonidine adrenoceptor inactivation, while these same ligands were only partially effective against SZL-49. Either SZL-49 or chlorethylclonidine significantly impaired the irreversible adrenoceptor blocking actions of phenoxybenzamine. These results suggest: (1) only the alpha 1a-adrenoceptor subtype appears to be associated with nerve terminals in the tail artery, (2) there may be a complex interaction between the alpha 1-adrenoceptor subtypes such that both receptors must be intact and functional to observe normal agonist and antagonist interactions, (3) there may be three sites of action for agonists associated with the rat aorta.
Asunto(s)
Receptores Adrenérgicos alfa/fisiología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Aorta Torácica/metabolismo , Unión Competitiva , Clonidina/análogos & derivados , Clonidina/antagonistas & inhibidores , Clonidina/farmacología , Dioxanos/metabolismo , Técnicas In Vitro , Cinética , Masculino , Metoxamina/metabolismo , Músculo Liso Vascular/fisiología , Músculo Liso Vascular/ultraestructura , Fenoxibenzamina/farmacología , Fenilefrina/antagonistas & inhibidores , Prazosina/análogos & derivados , Prazosina/antagonistas & inhibidores , Prazosina/metabolismo , Prazosina/farmacología , Ratas , Ratas Endogámicas , Receptores Adrenérgicos alfa/clasificaciónAsunto(s)
Nucleótidos de Citosina/metabolismo , Metoxamina/metabolismo , ARN de Transferencia/metabolismo , Tirosina/metabolismo , Autorradiografía , Secuencia de Bases , Electroforesis , Escherichia coli/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Nucleósidos/metabolismo , Páncreas , Isótopos de Fósforo , ARN Bacteriano/metabolismo , Ribonucleasas/metabolismoAsunto(s)
Asma/metabolismo , Epinefrina/farmacología , Hemodinámica/efectos de los fármacos , Isoproterenol/farmacología , Metoxamina/farmacología , Receptores de Droga , Adulto , Animales , Asma/tratamiento farmacológico , Asma/etiología , Glucemia/análisis , Presión Sanguínea/efectos de los fármacos , Gasto Cardíaco/efectos de los fármacos , Epinefrina/metabolismo , Ácidos Grasos no Esterificados/sangre , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Humanos , Infusiones Parenterales , Insulina/sangre , Isoproterenol/metabolismo , Metoxamina/metabolismo , Ratones , Persona de Mediana Edad , Sistema Nervioso Simpático/fisiología , Simpaticolíticos/fisiología , Resistencia Vascular/efectos de los fármacosRESUMEN
The present study determined the effect of 1 hour of in vivo ischemia on the response of the rabbit urinary bladder to specific autonomic agonists and on the intracellular adenosine triphosphate content. A 48 per cent decrease in the contractile response of the base to the alpha agonist methoxamine was noted. A 42 per cent decrease in the response of the bladder body to bethanechol was found. Neither tissue improved significantly after a 1-week recovery period. Ischemia caused an immediate 80 per cent decrease in the intracellular adenosine triphosphate content that improved to 50 per cent of control with a 1-week recovery period. Although the site of the defect is not definitely identified, acute ischemia resulted in a decreased contractile response that could result in bladder dysfunction.