RESUMEN
The potential involvement of catecholamines and in particular of alpha(2)-adrenoceptor-related signalling pathways, in the regulation of drug-metabolizing enzymes by stress was investigated in Wistar rats after exposure to the environmental pollutant benzo(alpha)pyrene. For this purpose, total cytochrome P450 content, the CYP1A2 mRNA levels, 7-methoxyresorufin-O-dealkylase (MROD), 7-pentoxyresorufin-O-dealkylase (PROD) and p-nitrophenol hydroxylase activity levels were determined in the livers of rats exposed to repeated restraint stress after treatment with benzo(alpha)pyrene coupled with pharmacological manipulations of peripheral and/or central catecholamines and alpha(2)-adrenoceptors. The data show that stress is a significant factor in the regulation of CYP1A2 induction and that catecholamines play a central role in the stress-mediated modulation of hepatic CYP1A2 inducibility by benzo(alpha)pyrene. The up-regulating effect of stress on benzo(alpha)pyrene-induced CYP1A2 gene expression was eliminated after a generalized catecholamine depletion with reserpine. Similarly, in a state where only peripheral catecholamines were depleted and central catecholamines remained intact after guanethidine administration, the up-regulating effect of stress was eliminated. It is apparent that stress up-regulates the induction of CYP1A2 by benzo(alpha)pyrene mainly via peripheral catecholamines, while central catecholamines hold a minor role in the regulation. Pharmacological manipulations of alpha(2)-adrenoceptors appear to interfere with the effect of stress on the regulation of CYP1A2 inducibility. Either blockade or stimulation of alpha(2)-adrenoceptors with atipamezole and dexmedetomidine respectively, eliminated the up-regulating effect of stress on CYP1A2 benzo(alpha)pyrene-induced expression, while it enhanced MROD activity. In contrast, stress and pharmacological manipulations of catecholamines and alpha(2)-adrenoceptors did not affect total P450 content, the CYP2B1/2-dependent PROD and the CYP2E1-dependent p-nitrophenol hydroxylase activities. In conclusion, stress is a significant factor in the regulation of the CYP1A2 inducibility by benzo(alpha)pyrene, which in turn is involved in the metabolism of a large spectrum of toxicants, drugs and carcinogenic agents. Although the mechanism underlying the stress effect on CYP1A2 induction has not been clearly elucidated, it appears that peripheral catecholamines hold a predominant role, while central catecholamines and in particular, central noradrenergic pathways hold a minor role.
Asunto(s)
Benzo(a)pireno/farmacología , Catecolaminas/metabolismo , Citocromo P-450 CYP1A2/biosíntesis , Contaminantes Ambientales/farmacología , Estrés Fisiológico/enzimología , Agonistas de Receptores Adrenérgicos alfa 2 , Agonistas alfa-Adrenérgicos/farmacología , Animales , Citocromo P-450 CYP1A2/genética , Dexmedetomidina/farmacología , Quimioterapia Combinada , Inducción Enzimática/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Guanetidina/farmacología , Imidazoles/farmacología , Inyecciones Intraperitoneales , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/enzimología , Ratas , Ratas Wistar , Receptores Adrenérgicos alfa 2/metabolismo , Reserpina/farmacología , Restricción FísicaRESUMEN
This study investigated the role of catecholamine-related signaling pathways in the regulation of hepatic cytochrome P450 (CYP2E1). Central and peripheral catecholamine depletion with reserpine down-regulated CYP2E1. On the other hand, selective peripheral catecholamine depletion with guanethidine increased CYP2E1 apoprotein levels. Enrichment of peripheral catecholamines with adrenaline suppressed p-nitrophenol hydroxylase activity (PNP). PNP activity was also markedly suppressed by l-DOPA. Stimulation of D(2)-receptors with bromocriptine up-regulated CYP2E1, as assessed by enzyme activity and protein levels, whereas blockade of D(2)-dopaminergic receptors with sulpiride down-regulated this isozyme. These findings indicate that central and peripheral catecholamines have different effects on CYP2E1. Central catecholamines appear related to the up-regulation, whereas the role of peripheral catecholamines is clearly related to the type and location of adrenoceptors involved. D(2)-receptor-linked signaling pathways have an up-regulating effect on CYP2E1, while D(1)-receptor pathways may down-regulate this isozyme. It is worth noting that the widespread environmental pollutant benzo(alpha)pyrene (B(alpha)P) altered the modulating effect of catecholaminergic systems on CYP2E1 regulation. In particular, whereas stimulation or blockade of adrenoceptors had no effect on constitutive PNP activity, exposure to B(alpha)P modified the impact of central and peripheral catecholamines and alpha(2)-adrenoceptors on CYP2E1 expression. It appears that under the influence of B(alpha)P, alpha(2)-adrenergic receptor-linked signaling pathways increased CYP2E1 apoprotein levels. Given that a wide range of xenobiotics and clinically used drugs are activated by CYP2E1 to toxic metabolites, including the production of reactive oxygen species (ROS), it is possible that therapies challenging dopaminergic receptor- and/or alpha(2)-adrenoceptor-linked signaling pathways may alter the expression of CYP2E1, thus affecting the progress and development of several pathologies.
Asunto(s)
Catecolaminas/farmacología , Citocromo P-450 CYP2E1/biosíntesis , Hígado , Receptores de Dopamina D2/metabolismo , Transducción de Señal , Animales , Benzo(a)pireno/farmacología , Western Blotting , Antagonistas de los Receptores de Dopamina D2 , Regulación hacia Abajo , Isoenzimas , Hígado/efectos de los fármacos , Hígado/enzimología , Hígado/metabolismo , Masculino , Ratas , Ratas Wistar , Receptores de Dopamina D2/agonistas , Transducción de Señal/efectos de los fármacos , Regulación hacia ArribaRESUMEN
CYP1A2, a principal catalyst for metabolism of various therapeutic drugs and carcinogens, among others, is in part regulated by the stress response. This study was designed to assess whether catecholamines and in particular adrenergic receptor-dependent pathways, modulate benzo(alpha)pyrene (B(alpha)P)-induced hepatic CYP1A2. To distinguish between the role of central and peripheral catecholamines in the regulation of CYP1A2 induction, the effect of central and peripheral catecholamine depletion using reserpine was compared to that of peripheral catecholamine depletion using guanethidine. The effects of peripheral adrenaline and L-DOPA administration were also assessed. The results suggest that alterations in central catecholamines modulate 7-methoxyresorufin O-demethylase activity (MROD), CYP1A2 mRNA and protein levels in the B(alpha)P-induced state. In particular, central catecholamine depletion, dexmedetomidine-induced inhibition of noradrenaline release and blockade of alpha(1)-adrenoceptors with prazosin, up-regulated CYP1A2 expression. Phenylephrine and dexmedetomidine-induced up-regulation may be mediated, in part, via peripheral alpha(1)- and alpha(2)-adrenoceptors, respectively. On the other hand, the L-DOPA-induced increase in central dopaminergic activity was not followed by any change in the up-regulation of CYP1A2 expression by B(alpha)P. Central noradrenergic systems appeared to counteract up-regulating factors, most likely via alpha(1)- and alpha(2)-adrenoceptors. In contrast, peripheral alpha- and beta-adrenoceptor-related signaling pathways are linked to up-regulating processes. The findings suggest that drugs that bind to adrenoceptors or affect central noradrenergic neurotransmission, as well as factors that challenge the adrenoceptor-linked signaling pathways may deregulate CYP1A2 induction. This, in turn, may result in drug-therapy and drug-toxicity complications.
Asunto(s)
Benzo(a)pireno/farmacología , Citocromo P-450 CYP1A2/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Receptores Adrenérgicos/metabolismo , Transducción de Señal , Inhibidores de Captación Adrenérgica/farmacología , Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Catecolaminas/metabolismo , Sistema Enzimático del Citocromo P-450/biosíntesis , Sistema Enzimático del Citocromo P-450/genética , Citocromos , Dexmedetomidina/farmacología , Epinefrina/farmacología , Guanetidina/farmacología , Levodopa/farmacología , Masculino , Oxidorreductasas/biosíntesis , Oxidorreductasas/genética , Prazosina/farmacología , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Reserpina/farmacología , Simpaticolíticos/farmacología , Regulación hacia ArribaRESUMEN
Alpha2-adrenoceptor agents as well as stress affect the activity of several hepatic monoxygenases including those related to CYP1A enzymes. This study was therefore designed to assess the role of central and/or peripheral catecholamines and, in particular, of adrenoceptors in the regulation of B(alpha)P-induced cytochrome CYP1A1 expression. In order to discriminate the role of central from that of peripheral catecholamines in the regulation of CYP1A1 induction, the effect of central and peripheral catecholamine depletion using reserpine versus only peripheral catecholamine depletion using guanethidine was assessed. By using selected agonists and antagonists, the role of alpha and beta-adrenoceptors in the regulation of CYP1A1 induction was evaluated. The results showed that the central catecholaminergic system has a negative regulatory effect on 7-ethoxyresorufin O-deethylase (EROD) inducibility by benzo(alpha)pyrene (B(alpha)P), and that this may be mediated via alpha1-, alpha2- and beta-adrenoceptors. Specifically, stimulation of alpha2-adrenoceptors with dexmedetomidine and blockade of alpha1- or beta-adrenoceptors with prazosin or propranolol respectively, resulted in a further increase of EROD inducibility. Adrenoceptors were found to be involved in the regulation of the CYP1A1 gene at mRNA level. Both, reduced noradrenaline release in central nervous system induced with dexmedetomidine and central catecholamine depletion, as well as blockade of central alpha1-adrenoceptors induced with prazosin, all were associated with up-regulation of CYP1A1 expression. In contrast, stimulation of central beta-adrenoceptors with isoprenaline resulted in a down-regulation of CYP1A1 expression. Our observations indicate that drugs, which stimulate or block adrenoceptors and catecholamine release may lead to complications in drug therapy and modulate the toxicity or carcinogenicity of drugs that are substrates for the CYP1A1.
Asunto(s)
Citocromo P-450 CYP1A1/biosíntesis , Regulación Enzimológica de la Expresión Génica/fisiología , Receptores Adrenérgicos/fisiología , Transducción de Señal/fisiología , Animales , Citocromo P-450 CYP1A1/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Reserpina/farmacología , Transducción de Señal/efectos de los fármacosRESUMEN
The present study investigated the involvement of catecholamines in stress-mediated alterations in CYP1A1 induction by benzo(alpha)pyrene (B(alpha)P) in Wistar rats. This was achieved by measuring EROD activity and CYP1A1 mRNA levels in liver tissue from rats exposed to restraint stress and B(alpha)P coupled with pharmacological modulation of peripheral and central catecholamine levels and different adrenoceptors. In a state of reserpine-induced central and peripheral catecholamine depletion, stress strongly suppressed EROD induction. Peripheral catecholamines do not appear to play a critical role in the stress-mediated modulation of EROD inducibility by B(alpha)P. Stress did not alter EROD inducibility by B(alpha)P when peripheral catecholamines were either depleted by guanethidine or supplemented by peripheral adrenaline administration. On the other hand, central noradrenergic systems appear to have a role in the stress-mediated changes in B(alpha)P-induced EROD activity and Cyp1A1 gene expression. Stimulation or blockade of noradrenaline release with atipamezole and dexmedetomidine, respectively, significantly modified the up-regulating effect of stress. Alpha1 adrenoceptors also appear to participate in the effect of stress on EROD inducibility. Alpha1-blockade with prazosin potentiated the up-regulating effect of stress, possibly preventing the down-regulating effect of noradrenaline. Beta adrenoceptors also seem to be involved directly or indirectly in the stress-mediated modulation of Cyp1A1, as propranolol (beta-antagonist) blocked the down-regulating effect of stress on B(alpha)P-induced Cyp1A1 gene expression. Plasma corticosterone alterations after stress were not related to alterations in the B(alpha)P-induced EROD activity and Cyp1A1 gene expression. In conclusion, stress appears to interfere in the regulation of B(alpha)P-induced hepatic CYP1A1 in an unpredictable manner and via signalling pathways not always directly related to catecholamines. In particular, whenever drug treatment disrupts noradrenergic neurotransmission, other stress-stimulated factors appear to modify the induction of CYP1A1. In summary, regulation of induction of hepatic CYP1A1 during stress appears to involve various components of the stress system, including central and peripheral catecholamines, which interact in a complex manner, yet to be elucidated.