Morphological changes induced by opioid receptor agonist treatment of B50 neuronal cells cultured in hypoxia

Abstract

Introduction: Hypoxia has been implicated in nerve cell deaths that occur in a variety of neurological disorders. Opioid receptor agonists have been shown to have some positive benefits on the nervous system. The aim of the present work was to investigate the effects of hypoxia and opioid receptor agonists' treatment on the morphology of B50 neuronal cell lines cultured in hypoxia. Materials and Methods: The B50 cells were cultured under normoxic conditions (21%O2; 5% CO2) as the control group and under hypoxic conditions (5%O2; 5% CO2) as the experimental group. Three opioid receptor agonists namely DAMGO (_) DSLET () and ICI-199,441 () were administered to the cells for 48 hours as treatment against hypoxia after 48 hours of culture at doses of 10 _M, 50 _M and 100 _M respectively. Neuronal morphology, viability, proliferation and differentiation were assessed using same field morphological assessment. In addition lactate dehydrogenase (LDH) leakage, cellular proliferation and DbcAMP induced differentiation were also assessed. _ opioid receptor mRNA was assessed using RT-PCR. Results: The results showed groups of dead and degenerating B50 neuronal cells and some significant changes (P<0.05) in cellular proliferation, viability cellular differentiation. The levels of LDH leakage showed normal B50 cells (100%), hypoxic cells (587%), and treated cells with 100 _M DAMGO (_) (143%), 50 _M DSLET () (140%) and 50 _M ICI-199,441 () (109%). The changes in morphology, LDH release, neuronal viability, proliferation and differentiation were shown to be dose-dependent between treated hypoxic B50 neurons in culture. Conclusion: The results indicate that opioid agonists have some potential benefits in the treatment of hypoxia-induced changes in neuronal B50 cells in culture.