Ferulic Acid exerts concentration-dependent anti-apoptotic and neuronal differentiation-inducing effects in PC12 and mouse neural stem cells

Ferulic Acid (FA) is a phenolic compound with anti-apoptotic and anti-oxidative properties. There are reports regarding its neuro-protective, neuro-proliferative and neuro-differentiative effects. However, effect of FA on neuronal differentiation and its effective neuro-protective and neuro-differentiative concentrations are unknown. Also the role of sirtuin molecules in neuroprotective effects of FA were not reported. We used PC12 and mouse neural stem cells (mNSCs) in our experiments. Intact and apoptotic (H2O2-exposed) cells were treated with different concentrations of FA, and then they were evaluated by MTT, quantitative real-time RT-PCR and immunostaining assays. FA treatment at low concentrations (50 mu g/ml) significantly reduced apoptosis in H2O2-treated PC12 cells. Real-time RT-PCR and western blot assays confirmed that FA induced this effect through stabilization and degradation of P53 by increasing the expression rate of SIRT1, SIRT7 and MDM2 but down-regulation of USP7. Beside this anti-apoptotic effect, treatments of PC12 cells and mNSCs with higher concentrations of FA (250-800 mu g/ml on PC12 cells and 100-500 mu g/ml on mNSCs) increased the rate of neuronal differentiation. Immunocytochemical staining for beta-tubulin III and Map2 verified the presence of mature neurons, and western blot assay showed that FA-treated PC12 cells had a stepwise rise of phosphorylated-ERK1/2 with increasing concentrations of FA. Our findings showed that FA at low concentrations has neuroprotective effect through up-regulation of SIRT1, SIRT7 and MDM2, and at higher concentrations can promote neural differentiation and neurite outgrowth.