Cerebrolysin protects PC12 cells from CoCl2-induced hypoxia employing GSK3β signaling

Cerebrolysin (EVER Neuro Pharma, Austria) is a peptidergic drug indicated for clinical use in stroke, traumatic brain injury and dementias. The therapeutic effect of Cerebrolysin is thought to ensure from its neurotrophic activity, which shares some properties with naturally occurring neurotrophic factors. However, the exact mechanism(s) of action of Cerebrolysin is yet to be fully deciphered. This study aimed to investigate the neuroprotective effect of Cerebrolysin in a widely used in vitro model of hypoxia-induced neuronal cytotoxicity, namely CoCl2-treatment of PC12 cells. CoCl2 cytotoxicity was indicated by a reduced cell-diameter, cell shrinkage, increased pro-apoptotic caspase-activities and a decreased metabolic activity. Cerebrolysin maintained the cell-diameter of CoCl2-treated naïve PC12 cells, decreased the activation of caspase 3/7 in naïve PC12 cells and restored the cells’ metabolic activity in naïve and differentiated PC12 cells. Cerebrolysin treatment also decreased the levels of superoxides observed after exposure to CoCl2. Investigating the mechanism of action, we could demonstrate that Cerebrolysin application to CoCl2-stressed PC12 cells increased the phosphorylation of GSK3ß, resulting in the inhibition of GSK3ß. This might become clinically relevant for Alzheimer’s disease, since GSK3ß activity has been linked to the production of amyloid beta. Taken together, Cerebrolysin was found to have neuroprotective effects in CoCl2-induced cytotoxicity in PC12 cells.