ABSTRACT
Aerial parts of Medicago sativa L. have been used as food and its consumption has been associated with health benefits, one among the most important being menopausal symptoms control. This work was aimed to explore possible pharmacological effects of two new alfalfa-derived products that have recently emerged as daily beverage preparations. In exploring their potential estrogenic effects, they produced no relevant alteration in the uterus. However, lowering glucose levels until normal values without causing further hypoglycemic effect were observed, when rats were treated with 1.5 g/kg/day samples. In vivo acute toxicity was not found when the alfalfa products were tested up to 3 g/kg rat weight. Furthermore, in vitro studies were conducted to assess their possible toxic effects. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase tests were carried out on the Caco-2 cell model to determine cell viability and membrane integrity. A concentration-dependent effect was observed, with a significant decrease in cell viability after exposure to concentrations of alfalfa product up to 100 mg/mL (after 3 h of incubation) and 50 mg/mL (after 24 h of treatment). Although in vitro level, the decrease in cell viability at these still low doses may underlie some toxicity, making necessary additional studies before any recommendation of a sustained consumption of these products by humans.
ABSTRACT
Solute neutral amino acid transporter 5 (SNAT5/SN2) is a member of the System N family, expressed in glial cells in the adult brain, able to transport glutamine, histidine or glycine among other substrates. Its tight association with synapses and its electroneutral mode of operation that allows the bidirectional movement of substrates, supports the idea that this transporter participates in the function of the glutamine-glutamate cycle between neurons and glia. Moreover, SNAT5/SN2 might contribute to the regulation of glycine concentration in glutamatergic synapses and, therefore, to the functioning of the N-methyl-d-aspartate (NMDA) subtype of glutamate receptors. Ontogenic maturation of these synapses occurs postnatally through the coordinate expression of a large number of receptors, transporters, structural and regulatory proteins that ensure the correct operation of the excitatory pathways in the central nervous system. Since the temporal pattern of expression of SNAT5/SN2 is unknown, we analyzed it by immunoblot and immunohistochemical techniques. Results indicate that the expression of SNAT5/SN2 is triggered between the second and third postnatal week in the cerebral cortex, in parallel to the expression of the vesicular glutamate transporter vGLUT1 and the glial glutamate transporter GLT1/EAAT2. In the cerebellum, this process occurs about one week later than in the cerebral cortex. Immunohistochemical staining of cortical sections shows that from postnatal day 14 to adulthood the transporter was expressed exclusively in glial cells. Our results are consistent with the idea that SNAT5/SN2 expression is coordinated with that of other proteins necessary for the operation of glutamatergic synapses and reinforce the existence of a regulatory cross-talk between neurons and glia that orchestrates the building up of these synapses.
