Amburana cearensis seed extracts protect PC-12 cells against toxicity induced by glutamate

ABSTRACT Amburana cearensis (Allemão) A.C. Sm., Fabaceae, has been widely studied for its medicinal activities. Many neurodegenerative disorders are caused by oxidative stress, mitochondrial dysfunction, excitotoxicity induced by glutamate and ultimately cell death. This study describes the chemical profile of the ethanolic, hexane, dichloromethane, and ethyl acetate extracts obtained from seeds of A. cearensis. The objective of this study was to investigate the chemical profile of extracts obtained from seeds of A. cearensis, as well as their cytotoxicity and neuroprotective effects in cultures of neural PC12 cells. Metabolite profile was performed by GC–MS. PC12 cells were treated with increasing concentrations of the extracts (0.01–2000 µg/ml) and the cell viability was analyzed after 24 and 72 h using an MTT test. For the excitotoxicity assay, PC12 cells were pre-treated with glutamate (1 mM) for 6 h and treated with increasing concentrations (0.1–1000 µg/ml) of the extracts. The chromatographic analysis of the extracts detected various compounds with antioxidant properties, with the majority of peaks corresponding to the isoflavone coumarin. Only the hexane extract showed toxicity after 72 h exposure at the highest concentration (1000 µg/ml). By contrast, all extracts increased the cellular viability of PC12 cells against the toxicity caused by glutamate. Therefore, the extracts from the seeds of A. cearensis showed no toxicity and have neuroprotective potential against neuronal damage induced by glutamate, which may be related to their antioxidant properties.

Autophagy protects against neural cell death induced by piperidine alkaloids present in Prosopis juliflora (Mesquite)

ABSTRACT Prosopis juliflora is a shrub that has been used to feed animals and humans. However, a synergistic action of piperidine alkaloids has been suggested to be responsible for neurotoxic damage observed in animals. We investigated the involvement of programmed cell death (PCD) and autophagy on the mechanism of cell death induced by a total extract (TAE) of alkaloids and fraction (F32) from P. juliflora leaves composed majoritary of juliprosopine in a model of neuron/glial cell co-culture. We saw that TAE (30 µg/mL) and F32 (7.5 µg/mL) induced reduction in ATP levels and changes in mitochondrial membrane potential at 12 h exposure. Moreover, TAE and F32 induced caspase-9 activation, nuclear condensation and neuronal death at 16 h exposure. After 4 h, they induced autophagy characterized by decreases of P62 protein level, increase of LC3II expression and increase in number of GFP-LC3 cells. Interestingly, we demonstrated that inhibition of autophagy by bafilomycin and vinblastine increased the cell death induced by TAE and autophagy induced by serum deprivation and rapamycin reduced cell death induced by F32 at 24 h. These results indicate that the mechanism neural cell death induced by these alkaloids involves PCD via caspase-9 activation and autophagy, which seems to be an important protective mechanism.

Toxicological and toxicogenetics evaluation of plant used in cattle food and in popular medicine: indigofera suffruticosa mill

A planta Indigofera suffruticosa Mill é uma leguminosa que ocorre em pastagens. É utilizada como forrageira pelo gado e como planta medicinal por populaçöes carentes. Neste trabalho, foi avaliada a capacidade dos extratos e hexânico de folhas na induçäo de aberraçöes cromossômicas em células de medula óssea de camundongos. Grupos de animais foram tratados intraperitonealmente com 3 diferentes concentraçöes do extrato aquoso, 0,312, 0,625 e 1,250 g/Kg p.c., e 3 diferentes concentraçöes do extrato hexânico, 0,625, 1,250 e 2,50 g/Kg p.c.. Os animais foram sacrificados 24 horas após o tratamento. Näo se observou diferença significativa na freqüência de aberraçöes cromossômicas entre os grupos tratados com o extrato aquoso (3,0; 3,33; 5,0%) e o grupo controle negativo (4,6%). Para o extrato hexânico, um discreto aumento (5,7%) foi observado apenas para a menor concentraçäo, quando comparado com o controle negativo (4,8%). Outros estudos säo necessários, ainda, para avaliaçäo de um possível efeito genotóxico desta planta