RESUMEN
OBJECTIVE: The pathogenesis of non-alcoholic steatohepatitis is still unclear. We have demonstrated previously that peroxisome proliferator activated receptor gamma (PPARγ) ligand protects against inflammation and fibrogenesis in experimental non-alcoholic steatohepatitis. We aim to elucidate the effect and the mechanism of PPARγ itself on nutritional fibrotic steatohepatitis in mice. METHODS: C57BL/6J mice were fed with methionine-choline deficient (MCD) diet for 8 weeks to induce fibrotic steatohepatitis. Mice fed the MCD diet were treated with adenovirus carrying PPARγ (Ad-PPARγ), Ad-PPARγ plus PPARγ agonist rosiglitazone, or PPARγ antagonist 2-chloro-5-nitrobenzaniliden (GW9662), respectively. The effects of up-regulation of PPARγ in the presence or absence of its agonist/or antagonist were assessed by comparing the severity of hepatic injury, activation of hepatic stellate cells and the expression of adiponectin, heme oxygenase-1, and fibrogenic related genes. RESULTS: Mice fed with MCD diet for 8 weeks showed severe hepatic injury including hepatic steatosis, inflammatory infiltration, and fibrosis. Administration of Ad-PPARγ significantly lowered serum alanine aminotransferase level and ameliorated hepatic steatosis, necroinflammation, and fibrosis. These effects were associated with enhanced expression of PPARγ, up-regulated expression of adiponectin and heme oxygenase-1, and down-regulated expression of tumor necrosis factor alpha, interleukin-6, α-smooth muscle actin, transforming growth factor beta 1, matrix metallopeptidase-2, and -9. Administration of GW9662 promoted the severity of liver histology. CONCLUSIONS: The present study provided evidences for the protective role of overexpressing PPARγ in ameliorating hepatic fibrosing steatohepatitis in mice. Modulation of PPARγ expression might serve as a therapeutic approach for fibrotic steatohepatitis.
Asunto(s)
Hígado Graso/prevención & control , Vectores Genéticos/administración & dosificación , PPAR gamma/biosíntesis , PPAR gamma/uso terapéutico , Adenoviridae/genética , Anilidas/administración & dosificación , Animales , Colina , Dieta , Hígado Graso/etiología , Hígado Graso/genética , Hígado Graso/metabolismo , Inflamación/genética , Inflamación/fisiopatología , Cirrosis Hepática/etiología , Cirrosis Hepática/genética , Cirrosis Hepática/metabolismo , Cirrosis Hepática/prevención & control , Cirrosis Hepática Experimental , Masculino , Metionina/deficiencia , Ratones , Ratones Endogámicos C57BL , PPAR gamma/administración & dosificación , PPAR gamma/genética , Distribución Aleatoria , Rosiglitazona , Tiazolidinedionas/administración & dosificación , Transfección , beta-Galactosidasa/administración & dosificación , beta-Galactosidasa/genéticaRESUMEN
The neural substrates and mechanisms mediating the antinociceptive effects of the endogenous bioactive lipid, N-palmitoylethanolamide (PEA), require further investigation. We investigated the effects of exogenous PEA administration into the anterior cingulate cortex (ACC), an important brain region linked with cognitive and affective modulation of pain, on formalin-evoked nociceptive behaviour in rats. Potential involvement of peroxisome proliferator-activated receptor isoforms (PPAR) α and γ or endocannabinoid-mediated entourage effects at cannabinoid1 (CB1) receptors or transient receptor potential subfamily V member 1 (TRPV1) in mediating the effects of PEA was also investigated. Intra-ACC administration of PEA significantly attenuated the first and early second phases of formalin-evoked nociceptive behaviour. This effect was attenuated by the CB1 receptor antagonist AM251, but not by the PPARα antagonist GW6471, the PPARγ antagonist GW9662, or the TRPV1 antagonist 5'-iodo resiniferatoxin. All antagonists, administered alone, significantly reduced formalin-evoked nociceptive behaviour, suggesting facilitatory/permissive roles for these receptors in the ACC in inflammatory pain. Post-mortem tissue analysis revealed a strong trend for increased levels of the endocannabinoid anandamide in the ACC of rats that received intra-ACC PEA. Expression of c-Fos, a marker of neuronal activity, was significantly reduced in the basolateral nucleus of the amygdala, but not in the central nucleus of the amygdala, the rostral ventromedial medulla or the dorsal horn of the spinal cord. In conclusion, these data indicate that PEA in the ACC can reduce inflammatory pain-related behaviour, possibly via AEA-induced activation of CB1 receptors and associated modulation of neuronal activity in the basolateral amygdala.
Asunto(s)
Etanolaminas/farmacología , Etanolaminas/uso terapéutico , Giro del Cíngulo/efectos de los fármacos , Dolor/tratamiento farmacológico , Ácidos Palmíticos/farmacología , Ácidos Palmíticos/uso terapéutico , Receptor Cannabinoide CB1/metabolismo , Amidas , Animales , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/uso terapéutico , Antagonistas de Receptores de Cannabinoides/farmacología , Antagonistas de Receptores de Cannabinoides/uso terapéutico , Estudios de Cohortes , Modelos Animales de Enfermedad , Diterpenos/uso terapéutico , Fijadores/toxicidad , Formaldehído/toxicidad , Giro del Cíngulo/fisiología , Locomoción/efectos de los fármacos , Masculino , Microdisección , Microinyecciones , PPAR gamma/administración & dosificación , Dolor/inducido químicamente , Dimensión del Dolor , Proteínas Proto-Oncogénicas c-fos/genética , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas , Ratas Sprague-Dawley , Receptor Cannabinoide CB1/genéticaAsunto(s)
Biomarcadores/sangre , Enfermedades Cardiovasculares/etiología , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Reproducción/efectos de los fármacos , Tiazolidinedionas/uso terapéutico , Enfermedades Cardiovasculares/epidemiología , Enfermedades Cardiovasculares/prevención & control , Femenino , Hormona Folículo Estimulante/sangre , Estudios de Seguimiento , Humanos , Incidencia , Hormona Luteinizante/sangre , PPAR gamma/administración & dosificación , PPAR gamma/uso terapéutico , Síndrome del Ovario Poliquístico/sangre , Síndrome del Ovario Poliquístico/complicaciones , Radioinmunoensayo , Factores de Riesgo , Rosiglitazona , Federación de Rusia/epidemiología , Testosterona/sangre , Tiazolidinedionas/administración & dosificación , Resultado del Tratamiento , Adulto JovenRESUMEN
Peroxisome proliferator activated receptor gamma (PPARγ) has been known for their anti-inflammatory effects. But the application of this molecule in implant-induced inflammation has not been clearly studied yet. Here, we determined in vivo anti-inflammatory and osteogenic effects of PPARγ coated dental implant in the rat mandible. We used chitosan gold nanoparticles (Ch-GNPs) as a non viral vector to carry PPARγ plasmid DNA. Ch-GNPs were conjugated with PPARγ plasmid DNA through a coacervation process. Conjugation was cast over titanium (Ti) implants (4.5 × 0.8 mm) by dipping, and implants were installed in rat mandibles. One, 2, 3, and 6 weeks post-implantation, mandibles were examined by microcomputed tomography (µCT), immunohistochemistry, hematoxylin & eosin, and tartrate resistance acid phosphatase (TRAP) staining. In vivo Ch-GNPs/PPARγcoated implants were associated with inhibition of implant induced inflammatory molecules interleukin-1ß and receptor activator of nuclear factor kappa-B ligand and enhanced expression of osteogenic molecules like bone morphogenetic protein 2 and 7 (BMP-2/-7) by up-regulating anti-oxidant molecules heme oxygenase-1. µCT demonstrated that PPARγ overexpression increased the density and volume of newly formed bone surrounding the implants compared to control (n = 4; p < 0.05). Also, PPARγ reduced the number of TRAP positive cells. These results support the view that PPARγ overexpression diminishes inflammation and enhances osteogenesis around the dental implants. Thus, implant coated with anti-inflammatory molecules could have a significant utilization for the preparation of new biomaterials and may serve as prosthetic materials in patients suffering from inflammatory bone disease.
Asunto(s)
Materiales Biocompatibles Revestidos/uso terapéutico , Implantación Dental Endoósea/instrumentación , Implantes Dentales , Materiales Dentales/uso terapéutico , Osteogénesis por Distracción/instrumentación , PPAR gamma/farmacología , Animales , Proteínas Morfogenéticas Óseas/biosíntesis , Tornillos Óseos , Quitosano , ADN Recombinante/administración & dosificación , Portadores de Fármacos , Reacción a Cuerpo Extraño/metabolismo , Reacción a Cuerpo Extraño/patología , Reacción a Cuerpo Extraño/prevención & control , Oro , Hemo Oxigenasa (Desciclizante)/biosíntesis , Inflamación/prevención & control , Interleucina-1beta/análisis , Ensayo de Materiales , Nanopartículas del Metal , FN-kappa B/análisis , Oseointegración/fisiología , Osteoclastos/metabolismo , PPAR gamma/administración & dosificación , Ratas , Ratas Sprague-Dawley , TitanioRESUMEN
Lutein is a carotenoid pigment present in fruits and vegetables that has anti-inflammatory and antitumor properties. In this study, we examined the effect of lutein on proliferation and survival-associated genes in prostate cancer (PC-3) cells. We found that in vitro culture of PC-3 cells with lutein induced mild decrease in proliferation that improved in combination treatment with peroxisome proliferator-activated receptor gamma (PPARγ) agonists and other chemotherapeutic agents. Flow cytometry analyses showed that lutein improved drug-induced cell cycle arrest and apoptosis in prostate cancer. Gene array and quantitative reverse transcription-polymerase chain reaction analyses showed that lutein altered the expression of growth and apoptosis-associated biomarker genes in PC-3 cells. These findings highlight that lutein modulates the expression of growth and survival-associated genes in prostate cancer cells.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Proliferación Celular/efectos de los fármacos , Luteína/farmacología , Proteínas Oncogénicas/efectos de los fármacos , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/genética , Animales , Apoptosis/efectos de los fármacos , Biomarcadores de Tumor/genética , Carotenoides/farmacología , Bovinos , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Citometría de Flujo , Frutas/química , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Luteína/administración & dosificación , Masculino , PPAR gamma/administración & dosificación , PPAR gamma/agonistas , Neoplasias de la Próstata/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Verduras/químicaRESUMEN
The Jagged-Notch signalling, plays a crucial role in cell differentiation. Angiogenesis, is regulated by VEGF, bFGF as well as by the free fatty acid metabolites , which are regulators of transcription factors such as peroxisome proliferation activating receptors (PPARs). The study analyzed the signalling pathways involved in the regulation of Jagged-1/Notch-4 expression in endothelial cells (HUVECs) in response to VEGF, bFGF and PPAR-gamma exogenous activator - ciglitazone. HUVECs were incubated with investigated substances for 24 hours, with or without the presence of the MAP-kinases inhibitors were used. Jagged-1 and Notch-4 gene expression was determined using quantitative Real-Time PCR. The Jagged-1/Notch-4 protein expression was compared by flow cytometry, when the phosphorylation-dependent activation of kinases was estimated by Western-blot method. The opposite effect of VEGF, bFGF, or ciglitazone on the Jagged-1/Notch-4 expression on HUVEC was connected with the different activation of MAPKs. Ciglitazone, activated p38 MAPK pathway and simultaneously inhibited phosphorylation of p42/44 MAPK. The pro-angiogenic: bFGF and VEGF, also activated the p38 MAPK, but they did not attenuate the p42/44 MAPK phosphorylation. Maintaining of the Jagged/Notch interactions by VEGF, when down-regulation by bFGF and ciglitazone, seems to be dependent on the different effect on p38 MAPK and p42/44 MAPK pathway regulation.
Asunto(s)
Proteínas de Unión al Calcio/genética , Regulación de la Expresión Génica , Péptidos y Proteínas de Señalización Intercelular/genética , Proteínas de la Membrana/genética , Neovascularización Fisiológica/fisiología , Proteínas Proto-Oncogénicas/genética , Receptores Notch/genética , Western Blotting , Células Cultivadas , Factor 2 de Crecimiento de Fibroblastos/administración & dosificación , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Citometría de Flujo , Humanos , Proteína Jagged-1 , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , PPAR gamma/administración & dosificación , PPAR gamma/metabolismo , Receptor Notch4 , Proteínas Serrate-Jagged , Transducción de Señal , Tiazolidinedionas/farmacología , Venas Umbilicales/metabolismo , Factor A de Crecimiento Endotelial Vascular/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
PURPOSE: To determine whether rosiglitazone-enriched diet offer protection in a classical model of liver ischemia-reperfusion injury in rats. METHODS: Two days before the experiment, rats were divided into 2 groups: Control Group (n=13) rats fed with standard diet; Rosi Group (n=13): rats fed with a powdered standard diet supplemented with rosiglitazone. The animals were submitted to liver ischemia-reperfusion by clamping the pedicle of median and left anterolateral lobes. After 1 hour of partial hepatic ischemia, the clamp was removed for reperfusion. After 2 or 24 hours (Control and Rosi Groups), blood was collected for enzymes and cytokines analysis. Ischemic and non-ischemic liver were collected for malondialdehyde analysis and histological assessment. Lungs were removed for tissue myeloperoxidase quantification. RESULTS: There were no statistical differences between groups for all analysed parameters. CONCLUSION: In this model, rosiglitazone-enriched diet did not protect liver against ischemia-reperfusion injury.
Asunto(s)
Suplementos Dietéticos , Hígado/irrigación sanguínea , PPAR gamma/administración & dosificación , Daño por Reperfusión/prevención & control , Tiazolidinedionas/administración & dosificación , Animales , Aspartato Aminotransferasas/sangre , Citocinas/sangre , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Hígado/efectos de los fármacos , Hígado/enzimología , Masculino , Ratas , Ratas Wistar , Daño por Reperfusión/patología , RosiglitazonaRESUMEN
PURPOSE: To determine whether rosiglitazone-enriched diet offer protection in a classical model of liver ischemia-reperfusion injury in rats. METHODS: Two days before the experiment, rats were divided into 2 groups: Control Group (n=13) rats fed with standard diet; Rosi Group (n=13): rats fed with a powdered standard diet supplemented with rosiglitazone. The animals were submitted to liver ischemia-reperfusion by clamping the pedicle of median and left anterolateral lobes. After 1 hour of partial hepatic ischemia, the clamp was removed for reperfusion. After 2 or 24 hours (Control and Rosi Groups), blood was collected for enzymes and cytokines analysis. Ischemic and non-ischemic liver were collected for malondialdehyde analysis and histological assessment. Lungs were removed for tissue myeloperoxidase quantification. RESULTS: There were no statistical differences between groups for all analysed parameters. CONCLUSION: In this model, rosiglitazone-enriched diet did not protect liver against ischemia-reperfusion injury.
OBJETIVO: Determinar se a dieta enriquecida com rosiglitazona oferece proteção em um modelo clássico de lesão de isquemia e reperfusão hepática em ratos. MÉTODOS: Dois dias antes do experimento, os ratos foram divididos em 2 grupos: Grupo Controle (n=13): ratos alimentados com dieta padrão; Grupo Rosi (n=13): ratos alimentados com dieta em pó padrão enriquecida com rosiglitazona. Os animais foram submetidos à isquemia e reperfusão hepática por clampeamento do pedículo dos lobos médio e anterolateral esquerdo. Após 1 hora de isquemia, o clampe foi removido para a reperfusão. Após 2 ou 24 horas (Grupos Controle e Rosi), o sangue foi coletado para análise de enzimas e citocinas. Os fígados isquêmico e não isquêmico foram coletados para análise de malondialdeído e avaliação histológica. Pulmões foram removidos para quantificação da mieloperoxidase tecidual. RESULTADOS: Não houve diferenças estatísticas entre grupos em todos os parâmetros analisados. CONCLUSÃO: Nesse modelo, a dieta enriquecida com rosiglitazona não protegeu contra a lesão de isquemia e reperfusão hepática.