Abstract
The aim of the present study was to elucidate the intracellular mechanisms that cause neuronal cell death following exposure to excitatory neurotransmitter‑induced neurotoxicity, neurotoxins and oxidative stress. Human SH‑SY5Y neuroblastoma cells were exposed to various stimuli, including glutamate, 6‑hydroxydopamine (6‑OHDA), and glucose oxidase, and cell viability was determined by MTT assay. Early apoptosis and necrosis were examined by Annexin V/propidium iodide double staining and flow cytometric analysis. Intracellular calcium ion concentration and mitochondrial membrane potential were assessed by Fluo‑3a and JC‑1 staining, respectively. In addition, protein expression of receptor‑interacting protein (RIP) kinase 1 and RIP kinase 3 were evaluated by western blotting. Glutamate, 6‑OHDA and glucose oxidase treatment decreased cell viability. Glutamate induced apoptosis and necrosis, whereas, 6‑OHDA induced cell necrosis and glucose oxidase induced apoptosis. Furthermore, glutamate, 6‑OHDA or glucose oxidase treatment significantly increased intracellular calcium concentrations (P<0.05). The effect of glutamate on mitochondrial membrane potential varied with high and low concentrations, whereas 6‑OHDA and glucose oxidase sig...Continue Reading
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