Asunto(s)
Antiinflamatorios/uso terapéutico , Accidente Cerebrovascular/tratamiento farmacológico , Tetraciclinas/uso terapéutico , Animales , Isquemia Encefálica/tratamiento farmacológico , Inhibidores Enzimáticos/uso terapéutico , Humanos , Minociclina/uso terapéutico , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Fármacos Neuroprotectores/uso terapéutico , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
Spreading depression (SD) is a wave of sustained depolarization challenging the energy metabolism of cells without causing irreversible damage. SD is a major mechanism of gene induction that takes place in cortical injury, including ischemia. We studied the role of oxygen radicals in SD-induced c-fos and cyclooxygenase-2 (COX-2) induction using transgenic (Tg) mice that overexpress copper/zinc-superoxide dismutase (SOD1). The frequency, amplitude and duration of SD waves were similar in the Tg mice and wild-type littermates. c-fos and COX-2 mRNAs were strongly induced 1 and 4 h after SD. The induction of both genes was slightly but significantly less at 4 h in the Tg mice. The results indicate that even a mild, noninjurious metabolic stimulation increases the concentration of oxygen radicals to the level that contributes to gene expression.
Asunto(s)
Corteza Cerebral/metabolismo , Depresión de Propagación Cortical/fisiología , Expresión Génica/fisiología , Genes fos/fisiología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Superóxido Dismutasa/genética , Animales , Ciclooxigenasa 2 , Humanos , Isoenzimas/metabolismo , Proteínas de la Membrana , Ratones , Ratones Transgénicos , ARN Mensajero/metabolismo , Superóxido Dismutasa-1RESUMEN
The only treatment of patients with acute ischemic stroke is thrombolytic therapy, which benefits only a fraction of stroke patients. Both human and experimental studies indicate that ischemic stroke involves secondary inflammation that significantly contributes to the outcome after ischemic insult. Minocycline is a semisynthetic second-generation tetracycline that exerts antiinflammatory effects that are completely separate from its antimicrobial action. Because tetracycline treatment is clinically well tolerated, we investigated whether minocycline protects against focal brain ischemia with a wide therapeutic window. Using a rat model of transient middle cerebral artery occlusion, we show that daily treatment with minocycline reduces cortical infarction volume by 76 +/- 22% when the treatment is started 12 h before ischemia and by 63 +/- 35% when started even 4 h after the onset of ischemia. The treatment inhibits morphological activation of microglia in the area adjacent to the infarction, inhibits induction of IL-1beta-converting enzyme, and reduces cyclooxygenase-2 expression and prostaglandin E(2) production. Minocycline had no effect on astrogliosis or spreading depression, a wave of ionic transients thought to contribute to enlargement of cortical infarction. Treatment with minocycline may act directly on brain cells, because cultured primary neurons were also salvaged from glutamate toxicity. Minocycline may represent a prototype of an antiinflammatory compound that provides protection against ischemic stroke and has a clinically relevant therapeutic window.
Asunto(s)
Isquemia Encefálica/prevención & control , Inflamación/tratamiento farmacológico , Minociclina/uso terapéutico , Animales , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/uso terapéutico , Secuencia de Bases , Isquemia Encefálica/enzimología , Ciclooxigenasa 2 , Cartilla de ADN , Dinoprostona/biosíntesis , Inmunohistoquímica , Inflamación/enzimología , Isoenzimas/efectos de los fármacos , Isoenzimas/metabolismo , Masculino , Minociclina/farmacología , Prostaglandina-Endoperóxido Sintasas/efectos de los fármacos , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas Sprague-Dawley , Ratas Wistar , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidoresRESUMEN
BACKGROUND AND PURPOSE: Plasma glucose and spreading depression (SD) are both determinants of brain ischemia. The purpose of this study was to examine whether plasma glucose affects SD-induced gene expression in the cortex. METHODS: SD was induced by topical application of KCl. Hyperglycemia and hypoglycemia were induced by intraperitoneal injection of glucose and insulin, respectively. The expression of c-fos, cyclooxygenase-2 (COX-2), protein kinase C-delta (PKCdelta), and heme oxygenase-1 (HO-1) was determined by in situ hybridization. RESULTS: SD alone induced expression of c-fos (by 340%), COX-2 (210%), HO-1 (470%), and PKCdelta (410%). Hypoglycemia (2.4+/-0.9 mmol/L) alone did not induce gene expression, and hyperglycemia (22.1+/-3.7 mmol/L) alone induced only c-fos by 42%. When hypoglycemia was induced 30 minutes before SD, c-fos induction was enhanced by 145%, but the induction of HO-1 and PKCdelta was reduced to 43% and 64%, respectively. When hyperglycemia was induced 30 minutes before SD, c-fos induction was enhanced by 388% and COX-2 expression by 53%, whereas the induction of PKCdelta and HO-1 was reduced to 54% and 51%, respectively. The frequency, amplitude, and duration of direct current potentials were unaltered in hyperglycemic SD animals, whereas in hypoglycemic animals the duration was increased by 47%. CONCLUSIONS: While SD induces expression of several genes, the availability of glucose regulates the extent of the gene induction. The effect of glucose is different on early-response genes (c-fos and COX-2) compared with late-response genes. Plasma glucose may contribute to neuronal damage partially by regulating gene expression.
Asunto(s)
Glucemia/fisiología , Isquemia Encefálica/fisiopatología , Depresión de Propagación Cortical/fisiología , Hiperglucemia/fisiopatología , Animales , Análisis de los Gases de la Sangre , Química Encefálica/fisiología , Isquemia Encefálica/enzimología , Isquemia Encefálica/genética , Ciclooxigenasa 2 , Electrofisiología , Regulación Enzimológica de la Expresión Génica , Hiperglucemia/enzimología , Hiperglucemia/genética , Inmunohistoquímica , Hibridación in Situ , Isoenzimas/análisis , Isoenzimas/genética , Masculino , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Peroxidasas/análisis , Peroxidasas/genética , Cloruro de Potasio/farmacología , Prostaglandina-Endoperóxido Sintasas/análisis , Prostaglandina-Endoperóxido Sintasas/genética , Proteínas Proto-Oncogénicas c-fos/análisis , Proteínas Proto-Oncogénicas c-fos/genética , ARN Mensajero/análisis , Ratas , Ratas Wistar , Activación TranscripcionalRESUMEN
Ischemic stroke is the most common life-threatening neurological disease and has limited therapeutic options. One component of ischemic neuronal death is inflammation. Here we show that doxycycline and minocycline, which are broad-spectrum antibiotics and have antiinflammatory effects independent of their antimicrobial activity, protect hippocampal neurons against global ischemia in gerbils. Minocycline increased the survival of CA1 pyramidal neurons from 10.5% to 77% when the treatment was started 12 h before ischemia and to 71% when the treatment was started 30 min after ischemia. The survival with corresponding pre- and posttreatment with doxycycline was 57% and 47%, respectively. Minocycline prevented completely the ischemia-induced activation of microglia and the appearance of NADPH-diaphorase reactive cells, but did not affect induction of glial acidic fibrillary protein, a marker of astrogliosis. Minocycline treatment for 4 days resulted in a 70% reduction in mRNA induction of interleukin-1beta-converting enzyme, a caspase that is induced in microglia after ischemia. Likewise, expression of inducible nitric oxide synthase mRNA was attenuated by 30% in minocycline-treated animals. Our results suggest that lipid-soluble tetracyclines, doxycycline and minocycline, inhibit inflammation and are neuroprotective against ischemic stroke, even when administered after the insult. Tetracycline derivatives may have a potential use also as antiischemic compounds in humans.
Asunto(s)
Hipocampo/patología , Ataque Isquémico Transitorio/prevención & control , Microglía/patología , Fármacos Neuroprotectores/farmacología , Células Piramidales/patología , Tetraciclinas/farmacología , Animales , Doxiciclina/farmacología , Gerbillinae , Proteína Ácida Fibrilar de la Glía/análisis , Proteína Ácida Fibrilar de la Glía/genética , Hipocampo/efectos de los fármacos , Humanos , Ataque Isquémico Transitorio/patología , Masculino , Microglía/efectos de los fármacos , Microglía/fisiología , Minociclina/farmacología , Células Piramidales/efectos de los fármacos , Células Piramidales/fisiología , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética/efectos de los fármacosRESUMEN
Repetitive spreading depression (SD) waves, involving depolarization of neurons and astrocytes and up-regulation of glucose consumption, is thought to lower the threshold of neuronal death during and immediately after ischemia. Using rat models for SD and focal ischemia we investigated the expression of cyclooxygenase-1 (COX-1), the constitutive form, and cyclooxygenase-2 (COX-2), the inducible form of a key enzyme in prostaglandin biosynthesis and the target enzymes for nonsteroidal anti-inflammatory drugs. Whereas COX-1 mRNA levels were undetectable and uninducible, COX-2 mRNA and protein levels were rapidly increased in the cortex, especially in layers 2 and 3 after SD and transient focal ischemia. The cortical induction was reduced by MK-801, an N-methyl-D-aspartic acid-receptor antagonist, and by dexamethasone and quinacrine, phospholipase A2 (PLA2) inhibiting compounds. MK-801 acted by blocking SD whereas treatment with PLA2 inhibitors preserved the wave propagation. NBQX, an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate-receptor antagonist, did not affect the SD-induced COX-2 expression, whereas COX-inhibitors indomethacin and diclofenac, as well as a NO synthase-inhibitor, NG-nitro-L-arginine methyl ester, tended to enhance the COX-2 mRNA expression. In addition, ischemia induced COX-2 expression in the hippocampal and perifocal striatal neurons and in endothelial cells. Thus, COX-2 is transiently induced after SD and focal ischemia by activation of N-methyl-D-aspartic acid-receptors and PLA2, most prominently in cortical neurons that are at a high risk to die after focal brain ischemia.
Asunto(s)
Corteza Cerebral/enzimología , Depresión de Propagación Cortical , Ataque Isquémico Transitorio/enzimología , Isoenzimas/biosíntesis , Neuronas/enzimología , Fosfolipasas A/metabolismo , Prostaglandina-Endoperóxido Sintasas/biosíntesis , Receptores de N-Metil-D-Aspartato/fisiología , Animales , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/fisiología , Cuerpo Estriado/enzimología , Depresión de Propagación Cortical/efectos de los fármacos , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Dexametasona/farmacología , Maleato de Dizocilpina/farmacología , Endotelio Vascular/enzimología , Inducción Enzimática , Inhibidores Enzimáticos/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Lateralidad Funcional , Hipocampo/enzimología , Inmunohistoquímica , Hibridación in Situ , Masculino , Proteínas de la Membrana , Neuronas/efectos de los fármacos , Neuronas/fisiología , Fosfolipasas A2 , Quinacrina/farmacología , Quinoxalinas/farmacología , ARN Mensajero/biosíntesis , Ratas , Ratas Wistar , Transcripción Genética/efectos de los fármacosRESUMEN
We used in situ hybridization to localize the long-term changes in ornithine decarboxylase (ODC) expression after a 90 min occlusion of the middle cerebral artery (MCAO) in the rat. The ODC mRNA was induced in the ipsilateral dentate gyrus (DG) and throughout the ischemic cortex at 12 h and still at 3 days after reperfusion. The induction was blocked by an N-methyl-D-aspartate (NMDA) receptor antagonist suggesting that ODC induction is NMDA receptor-mediated. The long-lasting up-regulation detected in regions where no cellular damage usually occurs, favors the hypothesis that ODC expression does not contribute to neuronal death after stroke.