Effect of aucubin on neural precursor cell survival during neuronal differentiation

Abstract

ABSTRACTPurpose of the study: Aucubin (ACB) is an iridoid glycoside with various biological activities. Previously, it is reported that ACB reduces cell survival and proliferation in many human tumors, whereas it facilitates cell survival and neuroprotection in damaged neuronal cells and disease models. However, its effects on cell survival in the non-proliferating or differentiated neurons are not known.Materials and methods: We examined whether ACB facilitated cell survival in differentiating neural precursor cells, HiB5, compared with the proliferating HiB5 cells at various concentrations.Results: The cell viabilities were evaluated by measuring MTT values, cell numbers, amounts of neurotransmittersD1 and protein amounts of neuronal markers. Here, we showed that ACB promotes cell survival in differentiated neurons (10–200 μg/mL), but reduces it in proliferating NPCs (200–400 μg/mL). Protein amounts of neurofilament proteins, NF-H, NF-M, PSD-95 in post-synaptic density, GAP-43 in growing neurites and NeuN in differentiated neurons were upregulated by addition of ACB, indicating that cell survival increased in differentiated neurons, shown by immunoblot analysis. Especially, when PDGF was added into N2 media to facilitate neuronal differentiation of HiB5 cells, the viability of differentiated HiB5 cells was significantly elevated following the increase of ACB concentration. Furthermore, ACB promoted cell survival of specific neuron types, such as GABAergic neurons and glutamatergic neurons. When differentiated neurons were immunostained with markers for specific neurons, neuronal subtypes producing GABA and GAD 65/67 were immunostained more than subtypes producing glutamate and vGluT1.Conclusion: These results indicate that ACB improves neuronal cell survival in differentiated cells, suggesting it may be a therapeutic compound for neurodegenerative disorders.