RESUMEN
Protein kinase CK2 ("casein kinase II") is a protein serine/threonine kinase that plays critical roles in biological processes such as cell growth, cell cycle progression, and apoptosis. So far, we have identified that one catalytic isozyme of CK2, CK2α, is over-expressed in the kidney during the progression of glomerulonephritis (GN). Moreover, we have shown that in vivo inhibition of CK2 by administration of CK2 inhibitors was effective in the treatment of experimental GN. Hence the development of potent CK2 inhibitors should be considered in therapeutic strategies for GN. In the present study we identified compound 13, a pyrazine derivative, as a potent CK2 inhibitor. By performing enzyme kinetics analysis in vitro, we characterized the inhibition of compound 13 toward each CK2 catalytic isozyme. Furthermore, in vivo, we demonstrated that compound 13 is effective in attenuating proteinuria, decreasing the enhanced level of blood urea nitrogen and serum creatinine, and ameliorating glomerular crescent formation in an experimental GN rat model. On the other hand, cellular apoptosis was detected in the rat testis following administration of compound 13. This study provides clues for new strategies for developing applicable compounds into CK2-targeted GN treatments.
Asunto(s)
Quinasa de la Caseína II/antagonistas & inhibidores , Glomerulonefritis/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/uso terapéutico , Animales , Apoptosis , Nitrógeno de la Urea Sanguínea , Quinasa de la Caseína II/metabolismo , Creatinina/sangre , Emodina/farmacología , Emodina/uso terapéutico , Glomerulonefritis/metabolismo , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/metabolismo , Glomérulos Renales/patología , Masculino , Inhibidores de Proteínas Quinasas/farmacología , Proteinuria , Pirazinas/farmacología , Pirazinas/uso terapéutico , Ratas Endogámicas WKY , Testículo/efectos de los fármacos , Testículo/patologíaRESUMEN
We identified 1-oxa-4,9-diazaspiro[5.5]undecane-based trisubstituted ureas as highly potent soluble epoxide hydrolase (sEH) inhibitors and orally active agents for treating chronic kidney diseases. Compound 19 exhibited excellent sEH inhibitory activity and bioavailability. When administered orally at 30 mg/kg, 19 lowered serum creatinine in a rat model of anti-glomerular basement membrane glomerulonephritis but 2,8-diazaspiro[4.5]decane-based trisubstituted ureas did not. These results suggest that 19 is an orally active drug candidate for treating chronic kidney diseases.
Asunto(s)
Alcanos/química , Descubrimiento de Drogas/métodos , Epóxido Hidrolasas/antagonistas & inhibidores , Insuficiencia Renal Crónica/enzimología , Urea/análogos & derivados , Administración Oral , Alcanos/administración & dosificación , Animales , Epóxido Hidrolasas/metabolismo , Humanos , Ratas , Insuficiencia Renal Crónica/tratamiento farmacológico , Solubilidad , Relación Estructura-Actividad , Urea/administración & dosificaciónRESUMEN
Oxidative stress is considered to be a key mechanism of hepatocellular injury and disease progression in patients with nonalcoholic steatohepatitis (NASH). The transcription factor Nrf2 (nuclear factor-erythroid-2-related factor 2) plays a central role in stimulating expression of various antioxidant-associated genes in the cellular defense against oxidative stress. As the cytosolic repressor kelch-like ECH-associated protein 1 (Keap1) negatively regulates Nrf2, activation of Nrf2 facilitated by its release from Keap1 may represent a promising strategy in the treatment of NASH. To test this hypothesis, we used two chemically distinct types of Nrf2 activator. One is the thiol-reactive agent oltipraz (OPZ), a typical Nrf2 activator, and the other is a novel biaryl urea compound, termed NK-252 (1-(5-(furan-2-yl)-1,3,4-oxadiazol-2-yl)-3-(pyridin-2-ylmethyl)urea). NK-252 exhibits a greater Nrf2-activating potential than OPZ. Furthermore, in vitro binding studies revealed that NK-252 interacts with the domain containing the Nrf2-binding site of Keap1, whereas OPZ does not. This finding indicates that NK-252 is more potent than OPZ due to its unique mechanism of action. For in vivo animal model studies, we used rats on a choline-deficient L-amino acid-defined (CDAA) diet, which demonstrate pathologic findings similar to those seen in human NASH. The administration of OPZ or NK-252 significantly attenuated the progression of histologic abnormalities in rats on a CDAA diet, especially hepatic fibrosis. In conclusion, by using Nrf2 activators with independent mechanisms of action, we show that, in a rat model of NASH, the activation of Nrf2 is responsible for the antifibrotic effects of these drugs. This strategy of Nrf2 activation presents new opportunities for treatment of NASH patients with hepatic fibrosis.
Asunto(s)
Hígado Graso/genética , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Aminoácidos/metabolismo , Animales , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Sitios de Unión/efectos de los fármacos , Sitios de Unión/genética , Línea Celular , Colina/metabolismo , Dieta , Dihidropiridinas/farmacología , Dioxinas/farmacología , Progresión de la Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Hígado Graso/metabolismo , Hígado Graso/patología , Fibrosis , Expresión Génica/efectos de los fármacos , Expresión Génica/genética , Humanos , Peróxido de Hidrógeno/farmacología , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch , Hígado/metabolismo , Masculino , NAD(P)H Deshidrogenasa (Quinona)/genética , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Enfermedad del Hígado Graso no Alcohólico , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Ratas , Transaminasas/sangre , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genéticaRESUMEN
We identified 2,8-diazaspiro[4.5]decane-based trisubstituted urea derivatives as highly potent soluble epoxide hydrolase (sEH) inhibitors and orally active agents for treating hypertension. Docking studies using human and murine sEH X-ray crystal structures revealed steric hindrance around the side chain of Phe406 of murine sEH. The trifluoromethyl moiety (11) was replaced with a trifluoromethoxy moiety (12) to prevent steric clash, and improved murine sEH inhibitory activity was observed. The oral administration of 12, 20, and 37 at a dose of 30mg/kg reduced blood pressure in spontaneously hypertensive rat, but had little effect on blood pressure in normotensive rat.
Asunto(s)
Antihipertensivos/química , Antihipertensivos/uso terapéutico , Epóxido Hidrolasas/antagonistas & inhibidores , Hipertensión/tratamiento farmacológico , Urea/análogos & derivados , Urea/uso terapéutico , Alcanos/química , Alcanos/farmacología , Alcanos/uso terapéutico , Animales , Antihipertensivos/farmacología , Presión Sanguínea/efectos de los fármacos , Epóxido Hidrolasas/química , Epóxido Hidrolasas/metabolismo , Humanos , Hipertensión/enzimología , Simulación del Acoplamiento Molecular , Ratas , Ratas Endogámicas SHR , Urea/farmacologíaRESUMEN
We report the discovery and structure-activity relationship of 2,6-disubstituted pyrazines, which are potent and selective CK2 inhibitors. Lead compound 1 was identified, and derivatives were prepared to develop potent inhibitory activity. As a result, we obtained compound 7, which was the smallest unit that retained potency. Then, introducing an aminoalkyl group at the 6-position of the indazole ring resulted in improved efficacy in both enzymatic and cell-based CK2 inhibition assays. Moreover, compound 13 showed selectivity against other kinases and in vivo efficacy in a rat nephritis model. These results show that 2,6-disubstituted pyrazines have potential as therapeutic agents for nephritis.
Asunto(s)
Quinasa de la Caseína II/antagonistas & inhibidores , Indazoles/química , Inhibidores de Proteínas Quinasas/química , Pirazinas/química , Animales , Sitios de Unión , Quinasa de la Caseína II/metabolismo , Simulación por Computador , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Humanos , Indazoles/síntesis química , Indazoles/uso terapéutico , Inyecciones Intraperitoneales , Nefritis/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/uso terapéutico , Estructura Terciaria de Proteína , Pirazinas/síntesis química , Pirazinas/uso terapéutico , Ratas , Relación Estructura-ActividadRESUMEN
Diabetes, a disease in which the body does not produce or use insulin properly, is a serious global health problem. Gut polypeptides secreted in response to food intake, such as glucagon-like peptide-1 (GLP-1), are potent incretin hormones that enhance the glucose-dependent secretion of insulin from pancreatic beta cells. Free fatty acids (FFAs) provide an important energy source and also act as signaling molecules in various cellular processes, including the secretion of gut incretin peptides. Here we show that a G-protein-coupled receptor, GPR120, which is abundantly expressed in intestine, functions as a receptor for unsaturated long-chain FFAs. Furthermore, we show that the stimulation of GPR120 by FFAs promotes the secretion of GLP-1 in vitro and in vivo, and increases circulating insulin. Because GLP-1 is the most potent insulinotropic incretin, our results indicate that GPR120-mediated GLP-1 secretion induced by dietary FFAs is important in the treatment of diabetes.
Asunto(s)
Ácidos Grasos no Esterificados/metabolismo , Glucagón/metabolismo , Mucosa Intestinal/metabolismo , Fragmentos de Péptidos/metabolismo , Precursores de Proteínas/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animales , Péptido 1 Similar al Glucagón , Humanos , Ratones , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Receptores Acoplados a Proteínas G/genéticaRESUMEN
As we had found previously that thapsigargin, an endomembrane Ca2+-ATPase inhibitor, induces production of intracellular platelet-activating factor (PAF) [Br. J. Pharmacol. 116 (1995) 2141], we decided to investigate the possible roles of intracellular PAF in nuclear factor (NF)-kappaB activation of thapsigargin-stimulated rat peritoneal macrophages. When rat peritoneal macrophages were stimulated with thapsigargin, the level of inhibitory protein of NF-kappaB-alpha (IkappaB-alpha) was decreased and the nuclear translocation of NF-kappaB was increased. The thapsigargin-induced activation of NF-kappaB was inhibited by the PAF synthesis inhibitor SK&F 98625 and the PAF antagonist E6123. Structurally unrelated PAF antagonists such as E5880 and L-652,731 also inhibited the thapsigargin-induced activation of NF-kappaB. Lipopolysaccharide (LPS)-induced activation of NF-kappaB was also suppressed by these drugs. In a culture of rat peritoneal macrophages, exogenously added PAF did not induce degradation of IkappaB-alpha. These findings suggest that the intracellular PAF produced by the stimulation with thapsigargin or LPS is involved in activation of the NF-kappaB pathway.
Asunto(s)
Macrófagos Peritoneales/metabolismo , FN-kappa B/metabolismo , Factor de Activación Plaquetaria/fisiología , Animales , Azepinas/farmacología , Secuencia de Bases , Células Cultivadas , Cartilla de ADN , Hidrólisis , Imidazoles/farmacología , Lipopolisacáridos/farmacología , Masculino , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Compuestos Organofosforados/farmacología , Fosforilación , Piperidinas/farmacología , Factor de Activación Plaquetaria/antagonistas & inhibidores , Compuestos de Piridinio/farmacología , Ratas , Ratas Sprague-Dawley , Tapsigargina/farmacología , Triazoles/farmacologíaRESUMEN
Incubation of rat peritoneal macrophages in the presence of thapsigargin increased production of prostaglandin E2, intracellular platelet-activating factor (PAF) and interleukin-6. However, no PAF was detected in the conditioned medium. In the presence of SK&F 98625 (diethyl 7-(3,4,5-triphenyl-2-oxo-2,3-dihydroimidazol-1-yl)heptane phosphonate), a CoA-independent transacylase inhibitor, the thapsigargin-induced increases in the interleukin-6 mRNA level and interleukin-6 production were suppressed in a concentration-dependent manner. This inhibitor also suppressed the production of prostaglandin E2 and intracellular PAF. The PAF receptor antagonists such as E6123 ((S)-(+)-6-(2-chlorophenyl)-3-cyclopropanecarbonyl-8,11-dimethyl-2,3,4,5-tetrahydro-8H-pyrido[4',3':4,5]thieno[3,2-f][1,2,4]triazolo [4,3-a][1,4]diazepine) and L-652,731 (2,5-bis(3,4,5-trimethylphenyl)tetrahydrofuran) partially inhibited the thapsigargin-induced increase in the levels of interleukin-6 mRNA and interleukin-6 protein. The SK&F 98625-induced suppression of interleukin-6 mRNA accumulation and interleukin-6 production was partially restored by addition of exogenous prostaglandin E2. However, exogenous PAF failed to reverse the suppression suggesting that the intracellular PAF does not act in an autocrine mechanism. These findings suggested that the concurrently produced prostaglandin E2 and intracellular PAF participate in the thapsigargin-induced increase in the interleukin-6 mRNA level and interleukin-6 production by rat peritoneal macrophages.
Asunto(s)
Dinoprostona/fisiología , Interleucina-6/biosíntesis , Factor de Activación Plaquetaria/fisiología , Receptores de Superficie Celular , Receptores Acoplados a Proteínas G , Tapsigargina/farmacología , Acetiltransferasas/antagonistas & inhibidores , S-Acetiltransferasa de la Proteína Transportadora de Grupos Acilo , Animales , Azepinas/farmacología , Células Cultivadas , Dinoprostona/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Inhibidores Enzimáticos/farmacología , Furanos/farmacología , Imidazoles/farmacología , Indometacina/farmacología , Interleucina-6/antagonistas & inhibidores , Interleucina-6/genética , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Masculino , Compuestos Organofosforados/farmacología , Factor de Activación Plaquetaria/antagonistas & inhibidores , Factor de Activación Plaquetaria/biosíntesis , Factor de Activación Plaquetaria/farmacología , Glicoproteínas de Membrana Plaquetaria/antagonistas & inhibidores , Glicoproteínas de Membrana Plaquetaria/fisiología , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Ratas , Ratas Sprague-Dawley , Tapsigargina/antagonistas & inhibidores , Triazoles/farmacologíaRESUMEN
Effects of beraprost sodium, a chemically stable prostacyclin analogue, on renal dysfunction in an experimental rat model of glomerulonephritis were investigated. Beraprost sodium (30, 100 and 300 microg/kg) was orally given twice daily from the late stage of nephritis in which renal dysfunction was already developed. Beraprost sodium treatment inhibited the increase in urinary protein, serum creatinine and blood urea nitrogen, and the decrease in creatinine clearance. The elevation of serum creatinine was also inhibited by predonisolone (1 mg/kg). However, captopril (25, 50 and 100 mg/kg) and dipyridamole (20 and 60 mg/kg) failed to inhibit the elevation of serum creatinine. In the beraprost sodium-treated nephritic rats, the increase in mRNA levels for monocyte chemoattractant protein-1 (MCP-1) and collagen in the kidney was inhibited. These results suggest that beraprost sodium ameliorates developed renal dysfunction and is possibly an effective agent for the treatment of human glomerulonephritis.
Asunto(s)
Epoprostenol/análogos & derivados , Epoprostenol/uso terapéutico , Glomerulonefritis/tratamiento farmacológico , Riñón/fisiopatología , Inhibidores de Agregación Plaquetaria/uso terapéutico , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Animales , Antiinflamatorios/uso terapéutico , Presión Sanguínea/efectos de los fármacos , Nitrógeno de la Urea Sanguínea , Peso Corporal/efectos de los fármacos , Captopril/uso terapéutico , Quimiocina CCL2/biosíntesis , Quimiocina CCL2/metabolismo , Creatinina/orina , Dipiridamol/uso terapéutico , Glomerulonefritis/fisiopatología , Riñón/efectos de los fármacos , Riñón/metabolismo , Pruebas de Función Renal , Glomérulos Renales/inmunología , Masculino , Prednisolona/uso terapéutico , Proteinuria/inducido químicamente , ARN Mensajero/biosíntesis , Conejos , Ratas , Ratas Endogámicas WKY , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The chloroform and the ethyl acetate fractions from the roots of Acanthopanax chiisanensis exhibited a significant inhibition of prostaglandin E2 (PGE2) production in rat peritoneal macrophages stimulated by the protein kinase C activator, 12-O-tetradecanoylphorbol 13-acetate (TPA). Hyperin was isolated as an active principle from the ethyl acetate fraction. It suppressed not only PGE2 production but also nitric oxide (NO) production in vitro in a concentration dependent manner, their IC50, being 24.3 and 32.9 microM, respectively. Hyperin also caused a significant inhibition of increase in acetic acid-induced vascular permeability in mice in vivo.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Eleutherococcus/química , Macrófagos Peritoneales/efectos de los fármacos , Quercetina/análogos & derivados , Quercetina/farmacología , Animales , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/aislamiento & purificación , Western Blotting , Permeabilidad Capilar/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Inyecciones Intraperitoneales , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/metabolismo , Masculino , Ratones , Óxido Nítrico Sintasa/metabolismo , Nitritos/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , Raíces de Plantas/química , Quercetina/química , Quercetina/aislamiento & purificación , Ratas , Ratas Sprague-DawleyRESUMEN
Beraprost sodium, a stable prostacyclin analog, was showed to improve survival rates in two different rat models, anti-glomerular basement membrane (GBM) glomerulonephritis (GN) and 5/6 nephrectomized (Nx) chronic kidney disease (CKD) rats. In the anti-GBM rat, beraprost sodium (0.2 and 0.6 mg/kg/day) improved survival rate (hazard ratio for beraprost sodium 0.6 mg/kg/day group, 0.10; 95% confidence interval, 0.01 to 0.68). Subsequently, in the 5/6 Nx CKD rat, beraprost sodium (0.6 mg/kg/day) improved survival rate (hazard ratio for beraprost sodium, 0.46; 95% confidence interval, 0.23 to 0.92), serum creatinine doubling time and the slope of the reciprocal of serum creatinine. In the anti-GBM GN rats, beraprost sodium suppressed the serum accumulation of representative uremic toxins such as indoxyl sulfate. Furthermore, beraprost sodium inhibited human aortic endothelial cell (HAEC) injury induced by indoxyl sulfate, indicating that beraprost sodium might have a protective effect against cardiovascular damage due to CKD. These results show that beraprost sodium can improve the survival rates in two rat models of anti-GBM GN and 5/6 Nx CKD rats by protecting endothelial cells and thereby ameliorating decreased renal function. Therefore, clinical studies are needed in patients with chronic kidney failure to determine whether beraprost sodium will become a useful medication in CKD.
Asunto(s)
Epoprostenol/análogos & derivados , Membrana Basal Glomerular/efectos de los fármacos , Glomerulonefritis/tratamiento farmacológico , Nefrectomía , Insuficiencia Renal Crónica/tratamiento farmacológico , Insuficiencia Renal Crónica/cirugía , Animales , Aorta/citología , AMP Cíclico/metabolismo , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Epoprostenol/farmacología , Epoprostenol/uso terapéutico , Glomerulonefritis/sangre , Humanos , Indicán/sangre , Masculino , Ratas , Insuficiencia Renal Crónica/sangre , Análisis de SupervivenciaAsunto(s)
Antineoplásicos Hormonales/farmacología , Apoptosis , Dexametasona/farmacología , Glucocorticoides/farmacología , Leucemia de Células B/enzimología , Serina Endopeptidasas/fisiología , Antineoplásicos Hormonales/antagonistas & inhibidores , Cumarinas/farmacología , Dexametasona/antagonistas & inhibidores , Perfilación de la Expresión Génica , Genes bcl-2 , Glucocorticoides/antagonistas & inhibidores , Granzimas , Humanos , Isocumarinas , Leucemia de Células B/genética , Leucemia de Células B/patología , Modelos Biológicos , ARN Mensajero/biosíntesis , Serina Endopeptidasas/genética , Inhibidores de Serina Proteinasa/farmacología , Transfección , Células Tumorales CultivadasRESUMEN
Glomerulonephritis (GN) is a progressive inflammation that may be caused by a variety of underlying disorders. It is the primary cause of chronic renal failure and end-stage renal disease, which require dialysis and transplantation worldwide. Immunosuppressive therapy has been used to treat GN clinically, but this treatment has had insufficient therapeutic effects. Here, we show that protein kinase CK2 is a key molecule in the progression of GN. cDNA microarray analysis identified CK2alpha, the catalytic subunit of CK2, as a GN-related, differentially expressed gene. Overexpression of CK2alpha was noted in the proliferative glomerular lesions in rat GN models and in renal biopsy specimens from lupus nephritis or IgA nephropathy patients. Administration of either antisense oligodeoxynucleotide against CK2alpha or low molecular weight CK2-specific inhibitors effectively prevented the progression of renal pathology in the rat GN models. The resolution of GN by CK2 inhibition may result from its suppression of extracellular signal-regulated kinase-mediated cell proliferation, and its suppression of inflammatory and fibrotic processes that are enhanced in GN. Our results show that CK2 plays a critical role in the progression of immunogenic renal injury, and therefore, CK2 is a potential target for GN therapy.
Asunto(s)
Quinasa de la Caseína II/antagonistas & inhibidores , Glomerulonefritis/tratamiento farmacológico , Oligodesoxirribonucleótidos Antisentido/farmacología , Subunidades de Proteína/antagonistas & inhibidores , Análisis de Varianza , Animales , Apigenina/administración & dosificación , Apigenina/farmacología , Apigenina/uso terapéutico , Nitrógeno de la Urea Sanguínea , Western Blotting , Quinasa de la Caseína II/metabolismo , Creatinina/sangre , Emodina/administración & dosificación , Emodina/farmacología , Emodina/uso terapéutico , Sueros Inmunes/administración & dosificación , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Riñón/patología , Masculino , Oligodesoxirribonucleótidos Antisentido/administración & dosificación , Oligodesoxirribonucleótidos Antisentido/uso terapéutico , Análisis de Secuencia por Matrices de Oligonucleótidos , Prednisolona , Subunidades de Proteína/metabolismo , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Organismos Libres de Patógenos EspecíficosRESUMEN
We analyzed gene expression in rat anti-Thy1 antibody-induced glomerulonephritis by using the cDNA microarray method. Ninety-seven genes that differed by more than 1.5-fold intensity in comparison with the controls were selected. Cluster analysis showed that the expression of genes associated with inflammation reached maximum levels at 24 h, while genes involved in the development of fibrosis increased at 7 days after injection. Microarray analysis of animal disease models may be a powerful approach for understanding the gene expression programs that underlie these disorders.
Asunto(s)
Perfilación de la Expresión Génica/métodos , Glomerulonefritis Membranoproliferativa/genética , Animales , Femenino , Perfilación de la Expresión Génica/estadística & datos numéricos , Glomerulonefritis Membranoproliferativa/patología , Análisis por Micromatrices/métodos , Análisis por Micromatrices/estadística & datos numéricos , Ratas , Ratas WistarRESUMEN
BACKGROUND/AIMS: When a loss of hepatic mass occurs, the expression of a large number of genes is either induced or altered, accompanying hepatocyte proliferation. In the present study, we made an in-house cDNA microarray containing 4608 elements (Liver chip), and analyzed extensively gene expression profiles of the regenerating liver after 70% partial hepatectomy (PHx) in rats. METHODS: RNAs were prepared from three rat livers at each time point (taken at 0, 6, 12, 18, 24, 48, 72 h, and 1 week after PHx). Using the liver chip, we performed large-scale analysis of gene expression during liver regeneration. Elements either up- or down-regulated more than twofold at one or more time points were selected. RESULTS: Among the 4608, 382 were identified. Using cluster analysis, we found great similarity between gene-expression profiles at 12 and 18 h after PHx as well as between 48 and 72 h after PHx. We also found that there are at least six distinct temporal patterns of gene expression in the regenerating rat liver after PHx. CONCLUSIONS: These results indicated that microarray analysis is a powerful approach for monitoring molecular events in the regenerating liver.