RESUMO
AIM: To evaluate the role of intestinal microflora in the effects of multi-herbal medicine on gene expression in the gut and liver. METHODS: The multi-herbal medicine Juzentaihoto (JTX) was administered to five germ-free mice and regular mice for 2 wk. Among the results of the comprehensive gene chip analysis of the intestine and liver, we featured heat shock proteins (HSPs) 70 and 105 because their gene expression changed only in the presence of microflora. Real-time RT-PCR was performed to confirm the expression levels of these HSP genes. To determine whether JTX acts directly on the HSP genes, sodium arsenite (SA) was used to induce the heat shock proteins directly. To examine the change of the intestinal microflora with administration of JTX, the terminal restriction fragment polymorphism (T-RFLP) method was used. To identify the changed bacteria, DNA sequencing was performed. RESULTS: Heat shock protein gene expression, documented by gene chip and real-time RT-PCR, changed with the administration of JTX in the regular mice but not in the germ-free mice. JTX did not suppress the direct induction of the HSPs by SA. T-RFLP suggested that JTX decreased unculturable bacteria and increased Lactobacillus johnsoni. These data suggested that JTX changed the intestinal microflora which, in turn, changed HSP gene expression. CONCLUSION: Intestinal microflora affects multi-herbal product JTX on the gene expression in the gut and liver.
Assuntos
Antineoplásicos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Proteínas de Choque Térmico/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/microbiologia , Fígado/metabolismo , Animais , Anti-Infecciosos/farmacologia , Arsenitos/farmacologia , Ciprofloxacina/farmacologia , Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP110/genética , Proteínas de Choque Térmico HSP110/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/genética , Lactobacillus/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos ICR , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Compostos de Sódio/farmacologiaRESUMO
One of the hypotheses regarding the pathogenesis of familial ALS (FALS) is a copper-mediated oxidative toxicity derived from the mutant Cu, Zn-superoxide dismutase (SOD1). We have previously demonstrated the efficacy of the combined treatment with a copper chelator (trientine) and an antioxidant (ascorbate) on the disease expression of the FALS-linked mutated SOD1 transgenic mice. Here, we investigated the efficacy of trientine or ascorbate alone on FALS mice when administered before or after the onset of the disease. The mice with a high dose of trientine or ascorbate administered before the onset survived significantly longer than the control. In the combined treatment with a high dose of trientine and ascorbate initiated before the onset, survival lengthened and the motor function of the mice remained more significantly than the control. None of the treatments affected the mean age of the onset, and none of the agents administered after the onset prolonged survival. These findings suggest that better outcomes may be expected by the administration of these agents at the preonset stage of the disease, and the combination of the agents acting on different sites might be useful in preserving the motor performance in FALS.
Assuntos
Esclerose Lateral Amiotrófica/tratamento farmacológico , Antioxidantes/uso terapêutico , Ácido Ascórbico/uso terapêutico , Quelantes/uso terapêutico , Trientina/uso terapêutico , Esclerose Lateral Amiotrófica/genética , Animais , Modelos Animais de Doenças , Progressão da Doença , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Quimioterapia Combinada , Humanos , Masculino , Camundongos , Camundongos Transgênicos , Superóxido Dismutase/genética , Superóxido Dismutase-1 , Taxa de Sobrevida , Resultado do TratamentoRESUMO
PURPOSE: Our previous studies showed that tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 are induced after hemorrhagic shock and that their induction is attenuated by hyperbaric oxygen treatment. In this study, we tried to identify the cell types in the liver that are responsible for the increase in TNF-alpha mRNA and IL-6 mRNA after hemorrhage using in situ reverse transcription-polymerase chain reaction (in situ RT-PCR). METHODS: Chronically cannulated rats were subjected to hemorrhage, maintaining a mean arterial blood pressure of 40 mmHg for 1 h. They were resuscitated with the collected blood and saline, and the livers were removed at designated times, then fixed and frozen. RESULTS: Standard in situ hybridization could not detect the mRNAs of TNF-alpha and IL-6 in the livers; however, in situ RT-PCR detected TNF-alpha mRNA and IL-6 mRNA mainly in the vascular endothelial cells and perivascular nonparenchymal cells of the bled rats. Sinusoidal cells were positive for TNF-alpha mRNA, but negative for IL-6 mRNA. No signal was found in normal rats, or when the experimental protocol was modified to: omit the RT step; precede the RT step with RNA digestion; or use an irrelevant probe. CONCLUSION: These results show that vascular endothelial cells and perivascular nonparenchymal cells are activated after massive hemorrhage to produce inflammatory cytokines.