benzyloxycarbonylvalyl-alanyl-aspartyl-fluoromethyl-ketone and 2-3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline

5-fluorouracil (5-FU) is a widely used anticancer drug. One of the adverse effects of this drug is selective cerebral white matter injury, but to the authors' knowledge its mechanism has not been well documented. The current study was performed to investigate the mechanism of cerebral white matter injury caused by 5-FU and to develop the intervention to attenuate its injury in vitro.. Mixed oligodendrocyte/astrocyte cells were dissociated from specimens taken from approximately 2-day-old postnatal mouse cortex and cultured for 3-4 weeks. The culture cells were exposed to 5-FU, cycloheximide, emetine, Z-VAD-fmk, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)-quinoxaline (NBQX), Trolox, and epigallocatechin gallate. Oligodendrocyte cell death was assessed by counting the number of viable galactocerebroside-positive cells per x 100 field.. Mixed oligodendrocyte/astrocyte culture cells that were exposed to 5-FU (at doses of 10 microM, 30 microM, and 100 microM) for 24 hours ensued concentration-dependent oligodendrocyte death. The majority of oligodendrocytes, but few astrocytes, were injured by 100 microM 5-FU. Trolox, a vitamin E analog antioxidant, as well as cycloheximide (a protein synthesis inhibitor) and Z-VAD-fmk (a caspase inhibitor), significantly attenuated the 5-FU-induced oligodendrocyte death. However, NBQX, an alpha-amino-2,3-dihydro-5-methyl-3-oxo-4-isoxazolepropionic acid (AMPA) receptor antagonist, did not appear to effect the 5-FU-induced oligodendrocyte death.. The findings of the current study suggested that 5-FU led to oligodendrocyte death rather than astrocyte death by way of the apoptotic process, whereas antioxidants may prevent the 5-FU-induced oligodendrocyte cell death in vitro.

Topics: Amino Acid Chloromethyl Ketones; Animals; Antimetabolites, Antineoplastic; Antioxidants; Caspase Inhibitors; Cell Culture Techniques; Cell Death; Chromans; Cycloheximide; Cysteine Proteinase Inhibitors; Excitatory Amino Acid Antagonists; Fluorouracil; Mice; Oligodendroglia; Protein Synthesis Inhibitors; Quinoxalines; Receptors, AMPA

The neurotoxic action of the abuse drugs methamphetamine (METH) and 3,4-methylenedioxymethamphetamine (MDMA) on cerebellar granule neurones (CGNs) culture was examined. Treatment for 48 h with METH or MDMA (1-5 mM) induced a higher decrease in viability than 24 h treatment. z.VAD.fmk (100 microM) but not MK-801 nor NBQX recovered control viability values. In both cases, cell death was characterised as apoptotic rather than necrotic by morphology cell observation. Apoptosis measured by flow cytometry indicated an increase in the hypodiploid population after 48 h treatment with METH and MDMA. Apoptosis was reverted by the presence of z.VAD.fmk (100 microM) but not by 10 microM MK-801 or NBQX. Similar results were obtained by analysing nuclear chromatine condensation. These results ruled out excitotoxic participation in amphetamine derivative-induced neurotoxicity in CGNs. Participation of radical oxygen species (ROS) was evaluated using alpha-tocopherol (1-15 microM) and cytometric studies. The co-treatment with 4 mM METH or MDMA for 48 h partially reverted neurotoxic action and apoptotic features, indicating ROS implication in CGNs death by amphetamine derivatives. Alteration of mitochondrial function induced cytochrome C (Cyt C) release after 48-h treatment with METH and MDMA (4 mM). There was also indication of caspase-3-like activation, measured by immunoanalysis and biochemically. Finally, neurodegenerative action caused by amphetamine derivatives may be prevented by using caspase inhibitors.

Topics: alpha-Tocopherol; Amino Acid Chloromethyl Ketones; Animals; Apoptosis; Caspase 3; Caspases; Cerebellum; Cytochromes c; Dizocilpine Maleate; Drug Interactions; Enzyme Activation; Flow Cytometry; Methamphetamine; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Mitochondria; N-Methyl-3,4-methylenedioxyamphetamine; Neurons; Neuroprotective Agents; Quinoxalines; Rats; Reactive Oxygen Species