RESUMO
Chitosan and Agaricus blazei Murill (ABM) extracts possess antitumor activities. The aim of the present study was to investigate whether chitosan, ABM extract or the two in combination were effective against tumors in tumorbearing mice. The mice were subcutaneously injected with SK-Hep 1 cells and were then were divided into the following six groups: Group 1, control group; group 2, chitosan 5 mg/kg/day; group 3, chitosan 20 mg/kg/day; group 4, ABM (246 mg/kg/day) and chitosan (5 mg/kg/day) combined; group 5, ABM (984 mg/kg/day) and chitosan (20 mg/kg/day) combined; and group 6, ABM (984 mg/kg/day). The mice were treated with the different concentrations of chitosan, ABM or combinations of the two for 6 weeks. The levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and vascular endothelial growth factor (VEGF), and tissue histopathological features were examined in the surviving animals. Based on the results of the investigation, the treatments performed in groups 2, 3 and 4 were identified as being capable of reducing the weights of the tumors, however, group 4, which was treated with chitosan (5 mg/kg/day) in combination with ABM (246 mg/kg/day) was able to reduce the levels of GOT and VEGF. As a result, treatment with chitosan in combination with ABM may offer potential in cancer therapy and requires further investigation.
Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Imunodeficiência Combinada Severa/tratamento farmacológico , Agaricus/química , Alanina Transaminase/biossíntese , Animais , Aspartato Aminotransferases/biossíntese , Carcinoma Hepatocelular/patologia , Quitosana/administração & dosagem , Quitosana/química , Modelos Animais de Doenças , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Hepáticas/patologia , Camundongos , Camundongos SCID , Oligossacarídeos/administração & dosagem , Oligossacarídeos/química , Extratos Vegetais/química , Imunodeficiência Combinada Severa/patologia , Fator A de Crescimento do Endotélio Vascular/biossínteseRESUMO
BACKGROUND: Despite growing claims of functional health benefits in folkloric medicine, the safety of chronic/elevated intakes of onion and garlic cannot be assumed. Therefore, this study assesses oral administration of varied doses of onion and garlic on some biomarkers of hepatic and renal functions in rats. METHODS: Animals were divided into five groups: control group received vehicle and extract-treated groups received varied doses of onion or garlic extract (0.5 mL and 1.0 mL/100 g bwt/day) for 6 weeks. RESULTS: Both doses of onion caused marked (p<0.05) increase in hepatic and renal levels of glutathione (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) and marked (p<0.05) decrease in malondialdehyde (MDA). Treatment with low dose of garlic elicited similar trend except in hepatic CAT, renal SOD and GST levels. A high dose of garlic only caused marked (p<0.05) increase in hepatic GST, renal GST, and SOD. Both doses of onion and low dose of garlic significantly (p<0.05) enhanced renal Na+/K+-ATPase activity. Only a high dose of onion caused significant (p<0.05) increase in hepatic aspartate transaminase (AST), alkaline phosphatase (ALP), and decrease in plasma AST activities. CONCLUSIONS: These findings suggest antioxidant enhancing capability for both doses of onion and low dose of garlic, while high dose of garlic elicited pro-oxidant conditions.
Assuntos
Alho , Rim/enzimologia , Fígado/enzimologia , Cebolas , Extratos Vegetais/farmacologia , Animais , Antioxidantes/metabolismo , Aspartato Aminotransferases/biossíntese , Biomarcadores , Catalase/biossíntese , Relação Dose-Resposta a Droga , Glutationa/biossíntese , Glutationa Transferase/biossíntese , Masculino , Malondialdeído/metabolismo , Ratos , Ratos Wistar , ATPase Trocadora de Sódio-Potássio/biossínteseRESUMO
Schwann cells have been identified as targets for glucocorticoids. Besides genes implicated in the myelination process, the target genes of glucocorticoids have not been identified in these cells. For that purpose, we performed microarray analysis on MSC80 (mouse Schwann cells) treated with a synthetic glucocorticoid, dexamethasone. These cells express a functional glucocorticoid receptor (GR), but none of the other steroid receptors. This allowed us to identify genes specifically regulated by GR in the absence of the mineralocorticoid receptor. Among the 5000 genes analyzed, 12 were at least two-fold upregulated and 91 genes were at least two-fold down-regulated upon treatment with dexamethasone. Because of their potential role in Schwann cell homeostasis, we selected, for further analysis, the upregulated genes encoding glutamine synthetase (GS) and cytosolic aspartate aminotransferase (cAspAT). These genes play a crucial role in the glutamate cycle which was shown to be vital in neuron-astrocyte cross-talk in the central nervous system. Their activation was confirmed by semi-quantitative and real-time PCR. A detailed analysis of cAspAT promoter activity revealed that the mechanism of regulation by GR in Schwann cells differs from that in hepatoma cells, suggesting a cell-specific regulation. The transactivation potency of the two Glucocorticoid Responsive Units (GRU) present in the cAspAT promoter seems to be dependent on the levels of the GR in MSC80 cells. Furthermore, we show that an increase in GR levels under certain circumstances could considerably potentiate the effects of glucocorticoids on the cAspAT promoter via synergistic activation of both GRU, To the opposite, an enhancement in GR levels did not further potentiate Dex-activation of the GS promoter, showing a differential mechanism of action of GR in the context of both promoters.
Assuntos
Aspartato Aminotransferases/genética , Citosol/enzimologia , Glucocorticoides/metabolismo , Glutamato-Amônia Ligase/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Animais , Aspartato Aminotransferases/biossíntese , Aspartato Aminotransferases/metabolismo , Astrócitos/metabolismo , Carcinoma Hepatocelular/metabolismo , Linhagem Celular , Cloranfenicol O-Acetiltransferase/metabolismo , DNA Complementar/metabolismo , Dexametasona/química , Dexametasona/farmacologia , Regulação para Baixo , Glutamato-Amônia Ligase/biossíntese , Glutamato-Amônia Ligase/metabolismo , Luciferases/metabolismo , Camundongos , Modelos Genéticos , Neurônios/metabolismo , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , RNA/metabolismo , Receptores de Glucocorticoides/metabolismo , Receptores de Mineralocorticoides/metabolismo , Elementos de Resposta , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células de Schwann/citologia , Ativação Transcricional , Transfecção , Regulação para CimaRESUMO
The effect of Himoliv (HV) was evaluated in carbon tetrachloride or paracetamol induced hepatotoxicity in rats. Liver necrosis was produced by administering single dose of either carbon tetrachloride (CCl4, 1 ml/kg, 50% v/v with olive oil, s.c.) or paracetamol (PC, 1 g/kg, p.o.). The liver damage was evidenced by elevated levels of serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and serum alkaline phosphatase (ALP) and hepatic thiobarbituric acid reacting substances (TBARS) and superoxide dismutase (SOD). HV pretreatment (0.5 and 1.0 ml/kg, p.o.) significantly (P < 0.001) reduced CCl4 or PC-induced elevations of the levels of SGOT, SGPT, ALP and TBARS, while the reduced concentration of SOD due to CCl4 or PC was reversed. Silymarin (25 mg/ kg, p.o.), a known hepatoprotective drug showed similar results.
Assuntos
Hepatopatias/tratamento farmacológico , Hepatopatias/enzimologia , Preparações de Plantas/uso terapêutico , Plantas Medicinais/química , Acetaminofen/antagonistas & inibidores , Acetaminofen/toxicidade , Alanina Transaminase/antagonistas & inibidores , Alanina Transaminase/biossíntese , Fosfatase Alcalina/antagonistas & inibidores , Fosfatase Alcalina/biossíntese , Animais , Aspartato Aminotransferases/antagonistas & inibidores , Aspartato Aminotransferases/biossíntese , Tetracloreto de Carbono/antagonistas & inibidores , Tetracloreto de Carbono/toxicidade , Doença Hepática Induzida por Substâncias e Drogas , Feminino , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Preparações de Plantas/farmacologia , Ratos , Ratos WistarRESUMO
Tea constituents that had a preventive effect on D-galactosamine-induced liver injury in rats were partially purified by column chromatography from a n-butanol-soluble fraction of green tea. The fraction containing glycosidic flavonoids was found to suppress the D-galactosamine-induced increase of plasma alanine aminotransferase and aspartate aminotransferase activities. These results indicate that glycosidic flavonoids contribute, at least in part, to the liver injury-preventive effect of green tea.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Flavonoides/uso terapêutico , Galactosamina , Chá/química , Alanina Transaminase/biossíntese , Animais , Aspartato Aminotransferases/biossíntese , Doença Hepática Induzida por Substâncias e Drogas/enzimologia , Cromatografia Líquida de Alta Pressão , Dieta , Flavonoides/química , Flavonoides/isolamento & purificação , Fígado/enzimologia , Masculino , Ratos , Ratos WistarRESUMO
To evaluate the benefit of green tea in mitigating hazards caused by repeated exposure of 2-nitropropane (2NP), we examined the effects of the tea on toxic indices, oxidative DNA damage and cell proliferation in the liver of 2NP-treated rats. Male Fischer 344 rats were administered, by gastric intubation, a total of six doses of 60 mg/kg 2NP(L), or alternatively two doses of 90 mg/kg and then four doses of 120 mg/kg 2NP(H) during 2 weeks. Green tea infusion was given to the rats as drinking water 1 week before the 2NP treatments and throughout the experiment. Significant elevation of hepatotoxic indices was evident in the 2NP(H)-treated group, such as an increase of serum glutamic-oxaloacetic transaminase (GOT) activity and of hepatic lipid peroxidation, together with a decrease in hepatic glycogen and serum triglyceride, and degenerative changes in the hepatocytes. A dose-related increase was observed in oxidative DNA damage and cell proliferation in the liver. Green tea effectively inhibited all of above changes induced by 2NP treatment, suggesting that tea intake may be effective for preventing the hepatic injuries after chronic exposure to 2NP.
Assuntos
Carcinógenos/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Dano ao DNA/efeitos dos fármacos , Neoplasias Hepáticas Experimentais/prevenção & controle , Fígado/efeitos dos fármacos , Nitroparafinas/toxicidade , Fitoterapia , Propano/análogos & derivados , Chá/uso terapêutico , Animais , Aspartato Aminotransferases/biossíntese , Divisão Celular/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Propano/toxicidade , Ratos , Ratos Endogâmicos F344RESUMO
Panicum miliaceum has at least three isozymes of aspartate aminotransferase (AspAT); the cytosolic and mitochondrial isozymes (cAspAT and mAspAT) are major components and the third is a minor isozyme. Fractionation of leaf subcellular components showed that the minor isozyme was localized in plastids (pAspAT). We purified the three isozymes from green leaves of P. miliaceum. Both cAspAT and pAspAT consisted of triple subforms having the same molecular size but different isoelectric points. No substantial difference in enzymatic properties was observed among these isozymes besides the pH profiles. We isolated a full-length cDNA clone for pAspAT. This clone contains an open reading frame that encodes 457 amino acids. The amino-terminal region of the pAspAT precursor shares common features of plastid transit peptides. The amino acid sequence of P. miliaceum pAspAT shows higher similarity with other plant pAspATs than P. miliaceum cAspAT and mAspAT. The mRNA levels of the three isozymes were high in leaves compared with roots and mesocotyls. The three isozymes showed different expression patterns against environmental stimuli such as light and nitrate. The activities and protein levels of cAspAT and mAspAT increased during greening in accordance with those of phosphoenolpyruvate carboxylase and NAD-malic enzyme involved in the C4 pathway, primarily as a consequence of the increase in the levels of their mRNAs. By contrast, pAspAT was constitutively expressed during greening. The activity and protein levels of cAspAT and mAspAT selectively increased during recovery from an nitrogen deficit, primarily as a consequence of increase in the levels of their mRNAs while those of pAspAT remained unchanged.
Assuntos
Aspartato Aminotransferases/metabolismo , Isoenzimas/metabolismo , Plantas/enzimologia , Sequência de Aminoácidos , Aspartato Aminotransferases/biossíntese , Aspartato Aminotransferases/química , Sequência de Bases , Cromatografia , Cromatografia em Gel , Cromatografia por Troca Iônica , Clonagem Molecular , DNA Complementar , Durapatita , Expressão Gênica , Isoenzimas/biossíntese , Isoenzimas/química , Cinética , Dados de Sequência Molecular , Peso Molecular , Folhas de Planta , Reação em Cadeia da PolimeraseRESUMO
The effect of pyridoxal depletion and supplementation on the intracellular level of mitochondrial and cytosolic aspartate aminotransferase in cultured chicken embryo fibroblasts was examined. No apoenzyme was detected in cells grown in the presence of pyridoxal, and the specific activity of total enzyme did not vary profoundly from primary to quaternary cultures. Under pyridoxal depletion, up to 40% apoenzyme was found in tertiary cultures which was entirely due to the mitochondrial isoenzyme. Cytosolic apoenzyme was never detected. Total aspartate aminotransferase relative to total protein was increased 2-fold in secondary cultures; only the mitochondrial isoenzyme contributed to the increased specific activity. The cytosolic isoenzyme decreased steadily and was below the limit of detection in quaternary cultures. The changes are attributed to an increased and decreased synthesis of mitochondrial and cytosolic isoenzyme, respectively. No induction of either isoenzyme was observed after incubating the cells with different hormones and substrates. In secondary cultures, no degradation of mitochondrial isoenzyme could be detected under pyridoxal deficiency or supplementation during 4.4 days, an interpassage duration. The cytosolic aspartate aminotransferase was degraded initially with an apparent half-life of approximately 0.9 day under both sets of conditions. The pronounced stability of mitochondrial aspartate aminotransferase, even though one-third of it was present as apoenzyme, excludes the formation of the apoform to be the rate-limiting step in its degradation. The present results show that pyridoxal affects the synthesis of mitochondrial and cytosolic aspartate aminotransferase, but differently.