RESUMEN
Sigma-1 receptors are ubiquitous multifunctional ligand-regulated molecular chaperones in the endoplasmic reticulum membrane with a unique history, structure, and pharmacological profile. Sigma-1 receptors bind ligands of different chemical structure and pharmacological action and modulate a wide range of cellular processes in health and disease, including Ca2+ signaling. To elucidate the involvement of sigma-1 receptors in the processes of Ca2+ signaling in macrophages we studied the effect of sigma-1 receptor ligands, phenothiazine neuroleptics chlorpromazine and trifluoperazine, on Ca2+ responses induced by inhibitors of endoplasmic Ca2+-ATPases thapsigargin and cyclopiazonic acid, as well as by disulfide-containing immunomodulators Glutoxim and Molixan in rat peritoneal macrophages. Using Fura-2AM microfluorimetry we showed for the first time that chlorpromazine and trifluoperazine inhibit both phases of Ca2+ responses induced by Glutoxim, Molixan, thapsigargin, and cyclopiazonic acid in rat peritoneal macrophages. The data obtained indicate the participation of sigma-1 receptors in a complex signaling cascade caused by Glutoxim or Molixan and leading to an increase in intracellular Ca2+ concentration in macrophages. The results also indicate the involvement of sigma-1 receptors in the regulation of store-dependent Ca2+entry in macrophages.
RESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that sigma-1 receptor antagonist neuroleptic chlorpromazine significantly inhibits glutoxim- and molixan-induced Ca2+ responses and Ca2+ responses induced by endoplasmic reticulum Са2+-ATPase inhibitors thapsigargin and cyclopiazonic acid in rat peritoneal macrophages. The results suggest the involvement of sigma-1 receptors in the signaling cascade induced by glutoxim or molixan and leading to intracellular Ca2+ concentration increase and in the regulation of store-dependent Ca2+ entry in macrophages.
Asunto(s)
Antipsicóticos/farmacología , ATPasas Transportadoras de Calcio/antagonistas & inhibidores , Calcio/metabolismo , Clorpromazina/farmacología , Retículo Endoplásmico/metabolismo , Macrófagos/metabolismo , Animales , Señalización del Calcio/efectos de los fármacos , Disulfuros/química , Combinación de Medicamentos , Indoles/farmacología , Inosina/farmacología , Macrófagos/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , Microscopía Fluorescente , Oligopéptidos/farmacología , Ratas , Ratas Wistar , Tapsigargina/farmacologíaRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that sigma-1 receptor agonist-tricyclic antidepressant amitriptyline-significantly inhibits store-dependent Ca2+ entry, induced by endoplasmic Ca2+-ATPase inhibitors thapsigargin and cyclopiazonic acid, in rat peritoneal macrophages. The results suggest a possible involvement of sigma-1 receptors in the regulation of store-dependent Ca2+ entry in macrophages.
Asunto(s)
Amitriptilina/farmacología , Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Macrófagos Peritoneales/metabolismo , Receptores sigma/agonistas , Animales , Indoles/farmacología , Transporte Iónico/efectos de los fármacos , Ratas , Ratas Wistar , Receptores sigma/metabolismo , Tapsigargina/farmacología , Receptor Sigma-1RESUMEN
Using voltage-clamp technique, the involvement of sigma-1 receptors in the regulation of Na+ transport in frog skin by the immunomodulatory drug glutoxim was investigated. We have shown for the first time that preincubation of the frog skin with the sigma-1 receptor antagonists haloperidol and chlorpromazine attenuates the stimulatory effect of glutoxim on the Na+ transport. The results suggest the possible involvement of the sigma-1 receptors in the regulation of Na+ transport in frog skin epithelium by glutoxim.
Asunto(s)
Proteínas Anfibias/antagonistas & inhibidores , Clorpromazina/farmacología , Haloperidol/farmacología , Oligopéptidos/farmacología , Receptores sigma/antagonistas & inhibidores , Piel/metabolismo , Sodio/metabolismo , Proteínas Anfibias/metabolismo , Animales , Transporte Iónico/efectos de los fármacos , Rana temporaria , Receptores sigma/metabolismo , Receptor Sigma-1RESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that sigma-1 receptor agonist, tricyclic antidepressant amitriptyline, significantly inhibits glutoxim- and molixan-induced Ca2+-responses in rat peritoneal macrophages. The results suggest possible involvement of sigma-1 receptors in the signaling cascade induced by glutoxim or molixan and leading to intracellular Ca2+ concentration increase in macrophages.
Asunto(s)
Amitriptilina/farmacología , Calcio/metabolismo , Inosina/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Oligopéptidos/farmacología , Animales , Combinación de Medicamentos , Ratas , Ratas Wistar , Receptores sigma/agonistas , Receptores sigma/metabolismo , Receptor Sigma-1RESUMEN
Using voltage-clamp technique, the involvement of epoxygenases in immunomodulatory drug glutoxim regulation of Na+ transport in frog skin was investigated. We have shown for the first time that preincubation of the frog skin with epoxygenase inhibitors econazole or proadifen almost completely inhibits the stimulatory effect of glutoxim on Na+ transport. The data suggest the involvement of the enzymes and/or products of epoxygenase oxidation pathway of arachidonic acid metabolism in glutoxim effect on Na+ transport in frog skin epithelium.
Asunto(s)
Econazol/farmacología , Inhibidores Enzimáticos/farmacología , Oligopéptidos , Oxidorreductasas/antagonistas & inhibidores , Piel/metabolismo , Sodio/metabolismo , Animales , Transporte Iónico/efectos de los fármacos , Oligopéptidos/farmacocinética , Oligopéptidos/farmacología , Rana temporariaRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that preincubation of macrophages with sigma-1 receptor antagonist haloperidol leads to a significant inhibition of the store-dependent Ca2+ entry induced by endoplasmic Ca2+-ATPase inhibitors thapsigargin or cyclopiazonic acid in rat peritoneal macrophages. The results suggest the involvement of the sigma-1 receptor in the regulation of storedependent Ca2+ entry in macrophages.
Asunto(s)
Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Haloperidol/farmacología , Macrófagos Peritoneales/metabolismo , Receptores sigma/antagonistas & inhibidores , Animales , Macrófagos Peritoneales/citología , Ratas , Receptor Sigma-1RESUMEN
Using Fura-2AM microfluorimetry, it was shown for the first time that phospholipase A2 inhibitors 4-bromophenacyl bromide and glucocorticosteroids prednisolone and dexamethasone attenuate Ca2+ responses induced by neuroleptic trifluoperazine in macrophages. The results suggest the involvement of phospholipase A2 and arachidonic acid metabolism cascade in the effect of trifluoperazine on intracellular Ca2+ concentration in macrophages.
Asunto(s)
Calcio/metabolismo , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Macrófagos/citología , Inhibidores de Fosfolipasa A2/farmacología , Trifluoperazina/farmacología , Animales , Ratas , Ratas WistarRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that preincubation of macrophages with the calsequestrin inhibitor neuroleptic trifluoperazine leads to a significant inhibition of the store-dependent Ca2+ entry induced by endoplasmic Ca2+-ATPase inhibitors thapsigargin or cyclopiazonic acid in rat peritoneal macrophages. The results suggest calsequestrin involvement in the regulation of the store-dependent Ca2+ entry in macrophages.
Asunto(s)
Calcio/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Trifluoperazina/farmacología , Animales , Transporte Biológico/efectos de los fármacos , Fura-2/análogos & derivados , Fura-2/metabolismo , Ratas , Ratas WistarRESUMEN
Using voltage-clamp technique, the involvement of lipoxygenases in the effect of immunomodulatory drug glutoxim on Na+ transport in frog skin was investigated. It was shown for the first time that preincubation of the skin with lipoxygenase inhibitors caffeic acid, baicalein, and nordihydroguaiaretic acid significantly decreases the stimulatory effect of glutoxim on Na+ transport. The data suggest the involvement of lipoxygenase oxidation pathway of arachidonic acid metabolism in the glutoxim effect on Na+ transport in frog skin epithelium.
Asunto(s)
Lipooxigenasas/metabolismo , Oligopéptidos/farmacología , Piel/efectos de los fármacos , Piel/metabolismo , Sodio/metabolismo , Animales , Anuros , Transporte Biológico/efectos de los fármacos , Epitelio/efectos de los fármacos , Epitelio/metabolismo , CinéticaRESUMEN
Using Fura-2AM microfluorimetry, it was shown for the first time that neuroleptic chlorpromazine causes intracellular Ca2+ concentration increase in macrophages due to Ca2+ mobilization from intracellular Ca2+ stores and subsequent Ca2+ entry from the external medium. Chlorpromazine-induced Ca2+ entry is inhibited by La3+ and 2-aminoethoxydiphenyl borate and is associated with Ca2+ store depletion.
Asunto(s)
Calcio/metabolismo , Clorpromazina/farmacología , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Macrófagos/citología , Animales , Relación Dosis-Respuesta a Droga , Ratas , Ratas WistarRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that preincubation of macrophages with methyl-ß-cyclodextrin, inducing cholesterol extraction from membranes and raft disruption, leads to significant inhibition of thapsigargin-induced store-dependent Ca2+ entry in rat peritoneal macrophages. In contrast, macrophage treatment with methyl-ß-cyclodextrin after Ca2+ entry mechanisms were activated by store depletion by thapsigargin application leads to potentiation of subsequent store-dependent Ca2+ entry. The results suggest that intact lipid rafts are necessary for the activation but not the maintenance of store-dependent Ca2+ entry in macrophages.
Asunto(s)
Calcio/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Tapsigargina/farmacología , beta-Ciclodextrinas/farmacología , Animales , Transporte Biológico/efectos de los fármacos , Interacciones Farmacológicas , RatasRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that sigma-1 receptor antagonist, antipsychotic haloperidol, significantly inhibits glutoxim- and molixan-induced Ca2+-response in peritoneal macrophages. These results indicate possible involvement of sigma-1 receptors in the signal cascade induced by glutoxim or molixan and leading to intracellular Ca2+ concentration increase in macrophages.
Asunto(s)
Antipsicóticos/farmacología , Señalización del Calcio , Haloperidol/farmacología , Inosina/farmacología , Macrófagos Peritoneales/metabolismo , Oligopéptidos/farmacología , Animales , Combinación de Medicamentos , Macrófagos Peritoneales/efectos de los fármacos , Ratas , Ratas Wistar , Receptores sigma/antagonistas & inhibidores , Receptor Sigma-1RESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that 5-lipoxygenase specific inhibitor antiasthmatic agent zileuton significantly inhibits Ca(2+)-responses induced by glutoxim and molixan in macrophages. The results support 5-lipoxygenase involvement in the effect of glutoxim and molixan on intracellular Ca(2+) concentration in macrophages and indicate the inadvisability of a combined use of drugs glutoxim and molixan and antiasthmatic agent zileuton.
Asunto(s)
Antiasmáticos/farmacología , Hidroxiurea/análogos & derivados , Inosina/farmacología , Inhibidores de la Lipooxigenasa/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Oligopéptidos/farmacología , Animales , Araquidonato 5-Lipooxigenasa/metabolismo , Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Señalización del Calcio/fisiología , Cationes Bivalentes/metabolismo , Células Cultivadas , Citofotometría , Combinación de Medicamentos , Interacciones Farmacológicas , Fura-2/análogos & derivados , Hidroxiurea/farmacología , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/enzimología , Macrófagos Peritoneales/enzimología , Microscopía Fluorescente , Ratas WistarRESUMEN
Using voltage-clamp technique, the involvement of WASP proteins and Arp2/3 complex in the effect of immunomodulator drug glutoxim on Na(+) transport in frog skin was investigated. It was shown for the first time that preincubation of the skin with the N-WASP inhibitor wiskostatin or the Arp2/3 complex inhibitor CK-0944666 significantly decreases the stimulatory effect of glutoxim on Na(+) transport. The data suggest the involvement of actin filament polymerization and branching in the glutoxim effect on Na(+) transport in frog skin.
Asunto(s)
Complejo 2-3 Proteico Relacionado con la Actina/antagonistas & inhibidores , Oligopéptidos/farmacología , Piel/efectos de los fármacos , Piel/metabolismo , Sodio/metabolismo , Proteína del Síndrome de Wiskott-Aldrich/antagonistas & inhibidores , Complejo 2-3 Proteico Relacionado con la Actina/metabolismo , Animales , Carbazoles/farmacología , Fármacos Dermatológicos/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Indoles/farmacología , Masculino , Técnicas de Placa-Clamp , Propanolaminas/farmacología , Rana temporaria , Proteína del Síndrome de Wiskott-Aldrich/metabolismoRESUMEN
Using Fura-2AM microfluorimetry, we have shown for the first time that methyl-ß-cyclodextrin, inducing cholesterol extraction from membranes and raft disruption, significantly inhibits glutoxim- and molixan-induced Ca2+-responses in rat peritoneal macrophages. The results suggest that intact rafts are necessary for signaling cascade induced by glutoxim or molixan and leading to intracellular Ca2+ concentration increase in macrophages.
Asunto(s)
Inosina/farmacología , Reguladores del Metabolismo de Lípidos/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Moduladores del Transporte de Membrana/farmacología , Oligopéptidos/farmacología , beta-Ciclodextrinas/farmacología , Animales , Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Señalización del Calcio/fisiología , Cationes Bivalentes/metabolismo , Células Cultivadas , Colesterol/metabolismo , Combinación de Medicamentos , Interacciones Farmacológicas , Colorantes Fluorescentes , Fluorometría , Fura-2/análogos & derivados , Macrófagos Peritoneales/metabolismo , Microdominios de Membrana/efectos de los fármacos , Microdominios de Membrana/metabolismo , Ratas WistarRESUMEN
Using Fura-2AM microfluorimetry the possible involvement of epoxygenase pathway of arachidonic acid metabolism in the effect of glutoxim and molixan on intracellular Ca2+ concentration in rat peritoneal macrophages was investigated. It was shown for the first time that preincubation of the macrophages with epoxygenase inhibitors, proadifen and econazole, significantly decreases the intracellular Ca2+ concentration increase induced by glutoxim and molixan. The addition of the epoxygenase inhibitors during the already developed store-dependent Ca(2+)-entry induced by glutoxim or molixan partially inhibits Ca(2+)-entry. The obtained data suggest the involvement of the products and/or enzymes of epoxygenase pathway of the arachidonic acid metabolism in the glutoxim and molixan effect on the Ca2+ signaling processes in macrophages.
Asunto(s)
Señalización del Calcio/efectos de los fármacos , Econazol/farmacología , Inhibidores Enzimáticos/farmacología , Macrófagos Peritoneales/metabolismo , Oligopéptidos/farmacología , Proadifeno/farmacología , Animales , Macrófagos Peritoneales/citología , Masculino , Ratas , Ratas WistarRESUMEN
The Fura-2AM fluorescent Ca(2+) probe was used to study the possibility that the Arp2/3 complex and WASP proteins are involved in the effects of glutoxim and molixan on the intracellular Ca(2+) concentration in macrophages. It has been demonstrated that preincubation of macrophages with inhibitors of the Arp2/3 complex or WASP proteins (CK-0944666 or wiskostatin, respectively) results in a significant suppression of Ca(2+)-responses induced by glutoxim or molixan. This suggests that polymerization of actin filaments is a process involved in the effect of glutoxim or molixan on intracellular Ca(2+) concentration in macrophages.
Asunto(s)
Complejo 2-3 Proteico Relacionado con la Actina/metabolismo , Calcio/metabolismo , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Familia de Proteínas del Síndrome de Wiskott-Aldrich/metabolismo , Complejo 2-3 Proteico Relacionado con la Actina/antagonistas & inhibidores , Animales , Carbazoles/farmacología , Cationes Bivalentes/metabolismo , Células Cultivadas , Combinación de Medicamentos , Colorantes Fluorescentes , Fura-2/análogos & derivados , Glutatión/análogos & derivados , Indoles/farmacología , Inosina/farmacología , Oligopéptidos/farmacología , Propanolaminas/farmacología , Ratas Wistar , Imagen de Colorante Sensible al Voltaje , Familia de Proteínas del Síndrome de Wiskott-Aldrich/antagonistas & inhibidoresRESUMEN
Using the fluorescent Ca(2+) probe Fura-2AM, the possible involvement of phospholipase A2, the key enzyme in the arachidonic acid cascade, in the effect of drugs glutoxim and molixan on the intracellular Ca(2+) concentration in macrophages was studied. It was shown for the first time that preincubation of macrophages with the classical phospholipase A2 inhibitor, 4-bromophenacyl bromide, as well as with glucocorticosteroids prednisolone and dexamethasone significantly inhibits Ca(2+) responses induced by glutoxim or molixan in macrophages. These results indicate the involvement of phospholipase A2 and arachidonic acid cascade in glutoximand molixan-induced signaling in macrophages.