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Olive virgin olive oil contain many phenolics effective against aging and neurodegeneration, particularly oleocanthal (OC) has been demonstrated to possess neuroprotective and anti-inflammatory activities. The aim of this work is to investigate the neuroprotective effect of OC in neuron-like SH-SY5Y cells before and after oxidative stress induced by H2O2. Using 2DE/MS the protein maps for different conditions have been obtained and analyzed. PCR analyses were performed to validate proteomic results. Thirty-three spots resulted differentially expressed after H2O2 insult, sixteen after treatment with OC followed by H2O2. Spots of interest were excised and identified by LC/MS/MS. OC significantly reverted the down-regulation induced by H2O2 of proteasome and ubiquitin proteins moreover it increased the expression of HSP90 and Protein DJ1. Moreover, OC was able to counteract oxidative stress induced by H2O2 in SH-SY5Y as measured by MTT viability assay and to increase reduced-GSH level. Our findings suggest that OC may have beneficial health effect in counteracting neurodegeneration.