RESUMO
The peripheral-type benzodiazepine receptor (PBR) is an 18-kDa high affinity drug- and cholesterol-binding protein that is involved in various cell functions, including cell proliferation and apoptosis. PBR was shown to be overexpressed in certain types of malignant human tumors and cancer cell lines, correlating with enhanced tumorigenicity and cell proliferation rates. The present study was conducted in order to further define the role of PBR in cancer and to extend our recent findings regarding the possible anticancer effects of the standardized Ginkgo biloba extract EGb 761. Treatment with EGb 761 decreased PBR mRNA levels and inhibited the proliferation of breast, glioma and hepatocarcinoma cell lines, further corroborating our previous contention that its mechanism of action is through the modification of PBR expression. In vivo treatment with Ginkgo biloba extract led to dose-dependent decreases in xenograft growth of both MDA-MB-231 breast cancer and U-87 glioma cell lines in nude mice, although the effects were not maintained after 50 days of treatment in the latter. The results obtained in MDA-MB-231 xenografts indicated pronounced inhibition of tumor growth, verified by MRI imaging. These results were obtained using a modified experimental protocol where the animals were treated with the extract before cell inoculation. Although an exact role for PBR in relation to the initiation and progression of various types of cancer remains to be defined, our results indicate that PBR overexpression in certain cancer cells is related to an aggressive phenotype. Since EGb 761 treatment opposes this aggressive phenotype by decreasing PBR overexpression, it could be useful in preventing or treating cancer invasiveness and metastasis.
Assuntos
Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Extratos Vegetais/farmacologia , Receptores de GABA/biossíntese , Animais , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Receptor alfa de Estrogênio/biossíntese , Receptor beta de Estrogênio/biossíntese , Feminino , Ginkgo biloba , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Nus , Neoplasias/genética , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ensaio Radioligante , Receptores de GABA/genética , Receptores de GABA/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Ginkgolide B (GKB) is a bioactive component of the standardized extract from the leaves of the Ginkgo biloba tree (EGb 761), which is used in Chinese and in occidental medicine. GKB is known as a platelet-activating factor receptor antagonist. Here, we provide evidence that GKB per se (0.25-5 microM) stimulated tyrosine phosphorylation of proteins, phospholipase D activation, calcium transients, and activation of p38 but not p44/42 Map kinases in human polymorphonuclear leukocytes (PMN). These stimulatory effects remained relatively weak and primed PMN for subsequent stimulation of respiratory burst (RB) or directed locomotion by the chemoattractant fMet-Leu-Phe (fMLP) or complement-derived factor C5a. A similar RB priming was observed with rat exudate PMN after in vivo administration of EGb 761 (25 and 50 mg/kg) to rats before pleurisy induction. Thus, GKB primarily induces activation of intracellular signaling events and has the potential to prime cellular functions such as PMN defense activities.
Assuntos
Diterpenos/administração & dosagem , Imunidade Inata/imunologia , Lactonas/administração & dosagem , Ativação de Neutrófilo/efeitos dos fármacos , Ativação de Neutrófilo/imunologia , Neutrófilos/imunologia , Pleurisia/imunologia , Pleurisia/prevenção & controle , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Ginkgolídeos , Humanos , Imunidade Inata/efeitos dos fármacos , Masculino , Neutrófilos/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologiaRESUMO
Ginkgo biloba extract (EGb 761) can improve cognitive function in patients with Alzheimer's disease, but the molecular mechanisms underlying this effect remain undefined. Because free cholesterol may be involved in the production of beta-amyloid precursor protein and amyloid beta-peptide, key events in the development of Alzheimer's disease, we examined EGb 761 in relation to cholesterol and amyloidogenesis. In aging rats, EGb 761 treatment lowered circulating free cholesterol and inhibited the production of brain beta-amyloid precursor protein and amyloid beta-peptide. Exposure of PC12 cells to EGb 761 decreased the processing of beta-amyloid precursor protein and abolished cholesterol-induced overproduction of this protein. Exposure of human NT2 cells to EGb 761 decreased free cholesterol influx and increased free cholesterol efflux. Our findings indicate that free circulating and intracellular cholesterol levels affect the processing of beta-amyloid precursor protein and amyloidogenesis. Our findings also provide the first demonstration that EGb 761 can influence these mechanisms.
Assuntos
Peptídeos beta-Amiloides/biossíntese , Precursor de Proteína beta-Amiloide/biossíntese , Colesterol/metabolismo , Extratos Vegetais/farmacologia , Envelhecimento , Doença de Alzheimer , Animais , Encéfalo/metabolismo , Linhagem Celular , Colesterol/sangue , Ginkgo biloba , Humanos , Lipoproteínas LDL/metabolismo , Masculino , Neurônios , Células PC12 , Ratos , Ratos Endogâmicos BN , Células-TroncoRESUMO
Two-dimensional electrophoresis (2D-PAGE) of metabolically labeled adrenocortical proteins, identified a series of spots at a molecular size of 90 kDa [isoelectric point (pI) 6.8-7.1; p90] that was induced by ACTH, but whose intensity was reduced in cells obtained from animals treated with an extract of Ginkgo biloba (EGb 761) and its purified component ginkgolide B (GKB). We have now identified p90. GKB (2 mg/kg x d, i.p.) was administered to rats for 8 d. Adrenocortical cells were prepared and stimulated with ACTH for 3 h. Cells obtained from saline-treated rats responded to ACTH by producing high amounts of corticosterone, an effect that was inhibited in cells obtained from GKB-treated animals. Samples were fractionated by 2D-PAGE and matrix-assisted laser desorption ionization mass spectrometry analysis of the p90 spots isolated from the gels revealed sequences sharing identity with the serotransferrin precursor protein. Further PCR screening of a rat adrenal cDNA library identified a sequence with a high degree of homology (79%) to serotransferrin precursor protein, and a lesser degree to rat transferrin (54%) and human melanotransferrin (32.8%). p90, in 2D-PAGE immunoblots, was also recognized by a monoclonal antibody raised against human 97-kDa melanotransferrin. Iron binding assays with rat adrenal cortex extracts further identified a 90-kDa melanotransferrin immunoreactive protein binding iron, suggesting that the identified protein, which we name "adrenotransferrin," may have iron-binding activity. This is the first report describing the presence of a serotransferrin precursor protein homolog belonging to the transferrin family and sharing epitopes with melanotransferrin in the adrenal, its induction by ACTH, and sensitivity to GKB.
Assuntos
Córtex Suprarrenal/metabolismo , Hormônio Adrenocorticotrópico/farmacologia , Diterpenos/farmacologia , Proteínas de Ligação ao Ferro/imunologia , Proteínas de Ligação ao Ferro/isolamento & purificação , Lactonas/farmacologia , Precursores de Proteínas , Precursores de Proteínas/imunologia , Precursores de Proteínas/isolamento & purificação , Transferrina/análogos & derivados , Córtex Suprarrenal/citologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Antígenos de Neoplasias , Células Cultivadas , Clonagem Molecular , Corticosterona/biossíntese , Ginkgolídeos , Ferro/metabolismo , Proteínas de Ligação ao Ferro/antagonistas & inibidores , Masculino , Antígenos Específicos de Melanoma , Dados de Sequência Molecular , Proteínas de Neoplasias/imunologia , Precursores de Proteínas/antagonistas & inibidores , Ratos , Ratos Sprague-DawleyRESUMO
Hippocampal neuron survival/growth and gene expression have been examined after prenatal (in utero) exposure of rats to EGb 761, a leaf extract of Ginkgo biloba. Oral administration of EGb 761 (100 or 300 mg/kg/day) to pregnant dams for 5 days increased the number of hippocampal neurons (maintained in culture) of their fetuses, indicating a neurotrophic effect of the extract. Using large-scale oligonucleotide microarrays containing over 8000 combined rat genes and expressed sequence tag clusters, it was shown that treatment of pregnant dams with EGb 761 (25, 50 or 100 mg/kg/day for 5 days) altered the expression of 187 genes in the hippocampi of male fetuses and 160 genes in those of female fetuses. Using gene-cluster analysis, these genes were grouped into 18 distinct clusters for males and 17 distinct clusters for females. Among these clusters, 35 genes shared a common expression pattern in male and female hippocampal development. Of these genes, the changes observed in insulin growth factor II, insulin growth factor binding protein 2, testosterone repressed prostate message-2, glutathione-dependent dehydroascorbate reductase, lipoprotein lipase, guanylate cyclase and DNA binding protein Brn-2 were confirmed by real-time quantitative polymerase chain reaction. These findings, which have provided the first genetic profile of the effects of EGb 761 on the developing rat hippocampus, increase our understanding of the molecular and genetic programs that are activated by the extract. These effects of EGb 761 may underlie its neuroprotective properties.
Assuntos
Sobrevivência Celular/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Extratos Vegetais/farmacologia , Efeitos Tardios da Exposição Pré-Natal , Animais , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Clusterina , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Relação Dose-Resposta a Droga , Embrião de Mamíferos , Feminino , Ginkgo biloba , Glicoproteínas/genética , Glicoproteínas/metabolismo , Guanilato Ciclase/genética , Guanilato Ciclase/metabolismo , Hipocampo/citologia , Hipocampo/embriologia , Hipocampo/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Fator de Crescimento Insulin-Like II/genética , Fator de Crescimento Insulin-Like II/metabolismo , Lipase Lipoproteica/genética , Lipase Lipoproteica/metabolismo , Masculino , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Família Multigênica , Neurônios/fisiologia , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Oxirredutases/genética , Oxirredutases/metabolismo , Gravidez , RNA Mensageiro/biossíntese , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
Recent studies conducted with various molecular, cellular and whole animal models have revealed that leaf extracts of Ginkgo biloba may have anticancer (chemopreventive) properties that are related to their antioxidant, anti-angiogenic and gene-regulatory actions. The antioxidant and associated anti-lipoperoxidative effects of Ginkgo extracts appear to involve both their flavonoid and terpenoid constituents. The anti-angiogenic activity of the extracts may involve their antioxidant activity and their ability to inhibit both inducible and endothelial forms of nitric oxide synthase. With regard to gene expression, a Ginkgo extract and one of its terpenoid constituents, ginkgolide B, inhibited the proliferation of a highly aggressive human breast cancer cell line and xenografts of this cell line in nude mice. cDNA microarray analyses have shown that exposure of human breast cancer cells to a Ginkgo extract altered the expression of genes that are involved in the regulation of cell proliferation, cell differentiation or apoptosis, and that exposure of human bladder cancer cells to a Ginkgo extract produced an adaptive transcriptional response that augments antioxidant status and inhibits DNA damage. In humans, Ginkgo extracts inhibit the formation of radiation-induced (chromosome-damaging) clastogenic factors and ultraviolet light-induced oxidative stress - effects that may also be associated with anticancer activity. Flavonoid and terpenoid constituents of Ginkgo extracts may act in a complementary manner to inhibit several carcinogenesis-related processes, and therefore the total extracts may be required for producing optimal effects.
Assuntos
Anticarcinógenos/farmacologia , Anticarcinógenos/uso terapêutico , Ginkgo biloba , Neoplasias/prevenção & controle , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Animais , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Ensaios Clínicos como Assunto , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias/irrigação sanguínea , Neoplasias/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Folhas de PlantaRESUMO
Treatment of rats and adrenocortical cells with ginkgolide B (GKB), a purified component of Ginkgo biloba leaf extracts, reduces the mRNA, protein, and ligand-binding levels of the adrenal peripheral-type benzodiazepine receptor (PBR), a mitochondrial cholesterol-binding protein, leading to decreased corticosteroid synthesis. In the Y1 adrenocortical cell line, GKB reduced both PBR levels and cyclic AMP-induced steroid formation. In these cells, GKB, but not various steroids and vitamins, reduced the expression of a reporter gene driven by the DNA sequence -624/-513 relative to the transcription start site of the PBR encoding gene. GKB treatment did not affect the SV40 promoter and increased the cytochrome P450 17alpha-hydroxylase gene promoter driven expression of the reporter gene. Electrophoretic mobility shift assays (EMSAs) indicated the presence of a functional transcriptional element bound to the -624/-513 DNA fragment. This GKB-induced inhibition of PBR was mediated by an interaction with a transcription factor that binds to the -636/-616 PBR-promoter region. Deletion or mutation of this sequence eliminated the DNA-protein interaction and the inhibitory effect of GKB on PBR gene transcription. This DNA-binding protein could be detected in nuclear extracts of rat brain, liver, and testis, but not kidney. It is also present in the human adrenal glands. However, the inhibitory effect following GKB treatment could be seen only in the adrenal glands. These results demonstrate that the GKB-activated inhibition of glucocorticoid production is due to a specific transcriptional suppression of the adrenal PBR gene and suggest that GKB might serve as a pharmacological tool to control excess glucocorticoid formation.
Assuntos
Córtex Suprarrenal/efeitos dos fármacos , Diterpenos , Lactonas/farmacologia , Receptores de GABA-A/genética , Esteroides/biossíntese , Transcrição Gênica/efeitos dos fármacos , Córtex Suprarrenal/fisiologia , Animais , Criança , Fibrinolíticos/farmacologia , Ginkgolídeos , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Ratos , Ratos Sprague-Dawley , Esteroides/antagonistas & inibidoresRESUMO
Myocardial stunning, a transient contractile dysfunction that appears following a brief period of ischemia, is at least partly due to the production of oxygen-derived free radicals. The objective of the present study was to determine whether the Ginkgo biloba extract EGb761, which has antioxidant properties in vitro, can attenuate myocardial stunning in vivo. Forty-seven anesthetized open-chest farm pigs underwent 10 min of occlusion of the left anterior descending coronary artery (LAD), followed by 3 hours of reperfusion. They were pretreated with either physiological saline, 100 mg or 300 mg of EGb 761 (Protocol I) or 3 mg or 9 mg of ginkgolide B (GkB) (Protocol II). Contractile function was assessed by sonomicrometry. Both doses of EGb 761 significantly improved recovery of contractile function in the reperfused myocardium with segment shortening averaging 23 +/- 5 % of baseline values at 3 hours post-reflow in controls versus 81 +/- 10 % and 57 +/- 12 % in the EGb100 and EGb300 groups, respectively (p < 0.05 vs control in both cases). In contrast, neither dose of GkB improved functional recovery during reperfusion. ESR experiments revealed that EGb761 resulted in a 59 % decrease in myocardial spin-adduct release during reperfusion (p < 0.05 versus control and GkB groups). A significant 28 % decrease (p < 0.05 vs control group) was also obtained in GkB-treated animals. These results indicate that EGb 761 can attenuate myocardial stunning following a brief ischemic insult in the in situ pig heart by an effect that involves a decrease in the formation of free radicals. As the effect of EGb 761 on functional recovery cannot be explained by the presence of GkB, the beneficial action of the extract on myocardial stunning likely involves complementary effects of both its non-ginkgolide and ginkgolide constituents.
Assuntos
Ginkgo biloba , Miocárdio Atordoado/tratamento farmacológico , Fitoterapia , Extratos Vegetais/farmacologia , Animais , Droperidol/administração & dosagem , Droperidol/farmacologia , Radicais Livres/metabolismo , Coração/efeitos dos fármacos , Coração/fisiologia , Hemodinâmica/efeitos dos fármacos , Injeções Intravenosas , Injeções Subcutâneas , Contração Miocárdica/efeitos dos fármacos , Isquemia Miocárdica/fisiopatologia , Miocárdio Atordoado/prevenção & controle , Tamanho do Órgão , Extratos Vegetais/uso terapêutico , Plantas Medicinais , SuínosRESUMO
Identification of the molecular switch controlling glucocorticoid synthesis might facilitate the development of pharmacological tools to control circulating cortisol levels. The transport of cholesterol from intracellular sources to the inner mitochondrial membrane represents the rate-determining step in the cascade of reactions leading to cortisol synthesis. A key element in this step is the peripheral-type benzodiazepine receptor (PBR). Several studies have indicated the beneficial effects of Ginkgo biloba on memory and stress control. Using pharmacological, biochemical and proteomic methods, we demonstrated that the standardized Ginkgo biloba extract EGb 761 and its isolated component ginkgolide B (GKB) inhibit PBR ligand binding and protein expression, resulting in decreased serum corticosterone levels. We further demonstrated that EGb 761- and GKB-induced inhibition of PBR protein is preceded by a decrease in mRNA-levels due to transcriptional suppression of PBR gene expression. Further studies indicated that the action of GKB is mediated by a transcription factor binding to the PBR gene promoter, thereby regulating PBR gene expression. These data indicate that EGb 761-induced inhibition of glucocorticoid production is due to specific transcriptional suppression of the adrenal PBR gene by GKB, and suggest that EGb 761 and GKB might serve as pharmacological tools to control excess glucocorticoid formation.
Assuntos
Diterpenos , Lactonas/farmacologia , Fármacos Neuroprotetores/farmacologia , Receptores de GABA-A/efeitos dos fármacos , Elementos de Resposta/genética , Transcrição Gênica/efeitos dos fármacos , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Ginkgolídeos , Extratos Vegetais/farmacologia , RatosRESUMO
The standardized extract of Ginkgo biloba leaves (EGb 761) has been shown to inhibit aggressive human breast cancer cell proliferation both in vitro and in vivo. These results were extended to human glioma and hepatoma cells in vitro suggesting that EGb 761 may have a more widespread application for tumor growth control. To understand the mechanism by which EGb 761 acts to inhibit cell proliferation, we investigated the effects of EGb 761 on human breast cancer, glioma and hepatoma cell transcriptomes by means of various large-scale DNA array techniques. The data presented focus on genes regulated by EGb 761 that are common to the three tumor cell types and for which the data were verified by two different types of DNA microarray and/or RNA (Northern) blot analysis and real-time quantitative PCR. These results could therefore help elucidate the mechanism of cytostatic action of EGb 761 and identify genes important for tumor growth.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Proteínas de Transporte/efeitos dos fármacos , Extratos Vegetais/farmacologia , Receptores de GABA/efeitos dos fármacos , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Ginkgo biloba , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Proteína II de Ligação a Poli(A)/efeitos dos fármacos , Células Tumorais CultivadasRESUMO
In this study, the effect of bilobalide, a purified terpene lactone component of the Ginkgo biloba extract (EGb 761), and EGb 761 against ischemic injury and against glutamate-induced excitotoxic neuronal death was compared. In the case of ischemic injury, neuronal loss and the levels of mitochondrial DNA (mtDNA)-encoded cytochrome oxidase (COX) subunit III mRNA in the hippocampal regions of gerbils was measured. A significant increase in neuronal death and a significant decrease in COX III mRNA were observed in the hippocampal CA1 neurons at 7-days of reperfusion after 5 min of transient global forebrain ischemia. Oral administration of EGb 761 at 25, 50 and 100 mg/kg/day and bilobalide at 3 and 6 mg/kg/day for 7 days before ischemia progressively protected hippocampal CA1 neurons against ischemia-induced neuronal death and reductions in COX III mRNA. In rat cerebellar neuronal cultures, addition of bilobalide or EGb 761 protected in a dose-dependent manner against glutamate-induced excitotoxic neuronal death [effective concentration (EC50) = 5 microg/ml (12 microM) forbilobalide and 100 microg/ml for EGb 761]. These results suggest thatboth EGb 761 and bilobalide protect against ischemia-induced neuronal death in vivo and glutamate-induced neuronal death in vitro by synergistic mechanisms involving anti-excitotoxicity, inhibition of free radical generation, scavenging of reactive oxygen species, and regulation of mitochondrial gene expression.
Assuntos
Ciclopentanos/farmacologia , Diterpenos , Furanos/farmacologia , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Administração Oral , Animais , Isquemia Encefálica/tratamento farmacológico , Morte Celular/efeitos dos fármacos , Cerebelo/efeitos dos fármacos , Ciclopentanos/administração & dosagem , Complexo IV da Cadeia de Transporte de Elétrons/biossíntese , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Furanos/administração & dosagem , Gerbillinae , Ginkgo biloba , Ginkgolídeos , Ácido Glutâmico/toxicidade , Hipocampo/efeitos dos fármacos , Masculino , Extratos Vegetais/farmacologia , Traumatismo por Reperfusão/tratamento farmacológicoRESUMO
Ginkgolide B (GKB, BN 52021) was described as a platelet-activating factor (Paf) receptor antagonist. However, it is not known whether all GKB biological effects are mediated through Paf receptor antagonism only. To gain insight into the drug mode of action, we investigated here the effects of GKB per se on functional and signaling activities in human polymorphonuclear leukocytes (PMN). Treatment of PMN with GKB (0.5-12 microM) stimulates a rapid and weak production of reactive oxygen species determined by chemiluminescence. ROS production required the activation of protein kinase C (PKC), tyrosine kinases and p38 mitogen-activated protein kinase as indicated by inhibitory effects of, respectively, GF 109203X (IC(50) of 0.5 microM), genistein (IC(50) of 0.5 microM) and SB 203580 (IC(50) of 0.2 microM) or SB 202190 (IC(50) of 1.1 microM). GKB stimulated a Pertussis toxin-sensitive PLD activity assessed by the formation of tritiated phosphatidic acid and choline. By contrast, GKB did prevent the Paf-mediated PLD activity and CL response (IC(50) of 2 microM). Interestingly, both GKB and Paf-induced CL response were prevented by selective Paf antagonists such as CV 6209 or WEB 2086 indicating that GKB may directly activate Paf receptors. Finally, GKB potentiated the CL response induced by fMet-Leu-Phe and zymosan. These results show that GKB is the first partial agonist of the Paf receptor described so far capable of priming the polymorphonuclear leukocyte function.
Assuntos
Diterpenos , Ginkgo biloba/química , Peptídeos e Proteínas de Sinalização Intracelular , Lactonas/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Glicoproteínas da Membrana de Plaquetas/metabolismo , Receptores de Superfície Celular , Receptores Acoplados a Proteínas G , Explosão Respiratória/efeitos dos fármacos , Azepinas/farmacologia , Proteínas de Transporte/farmacologia , Regulação para Baixo , Ativação Enzimática , Fibrinolíticos/farmacologia , Ginkgolídeos , Humanos , Técnicas In Vitro , Leucócitos Mononucleares/enzimologia , Leucócitos Mononucleares/fisiologia , Neutrófilos/fisiologia , Toxina Pertussis , Fosfolipase D/metabolismo , Extratos Vegetais/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Glicoproteínas da Membrana de Plaquetas/antagonistas & inibidores , Compostos de Piridínio/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Triazóis/farmacologia , Fatores de Virulência de Bordetella/farmacologiaRESUMO
In the present study, we investigated the effect of Ginkgo biloba extract, EGb 761, and one of its components, bilobalide, on gene expression of subunit 1 of mitochondrial NADH dehydrogenase (ND1) in PC12 cells. By Northern blot analysis we found a approximately 2-fold significant increase in NDI mRNA level, after 48 and 72 h exposure to 100 microg/ml EGb 761 and to 10 microg/ml bilobalide. We also evaluated, by oxygraphy measurements, mitochondrial respiration during state 3 and state 4. In cells treated with EGb 761 and bilobalide for 48 and 72 h, state 4 respiration was significantly decreased, and the respiratory control ratio was increased. These results provide evidence that EGb 761 and bilobalide exert their protective effects by up-regulating mitochondrial ND1 gene expression and by decreasing state 4 respiration, whose increase is thought to be responsible for oxidative damage.