Cannabinoid receptor agonists (CB1) have been shown to mediate several central nervous system (CNS) effects and actions, while the action of the G-protein coupled receptors like CB1 are involved in the activation of G-proteins necessary for several neural activities (Breivogel et al 2004; Croxford 2003; Smith 2002). The aim of this study was to investigate the role of Cannabinoid receptor agonists during neuronal hypoxia using cortical B50 cells in culture. The Cortical B50 cells were cultured in normal incubator (21%O2; 5% CO2)as the control group and hypoxic incubator (5%O2; 5% CO2)as the experimental group and three Cannabinoid agonists namely Win55,212-2 mesylate, Anandamide and 2-Arachidonylglycerol were selected and administered to the cells for 48 hours as treatment against hypoxia after 48 hours of culture at concentration of 10nM, 50nM and 100nM.Neuronal viability, proliferation, differentiation and second messenger activity were assessed using lactate dehydrogenase (LDH) leakage, cellular proliferation assay, second messenger (cAMP) assay, dbcAMP induced differentiation respectively and the levels of G-protein coupled receptors (CB1), mRNAs were assessed using RT-PCR. The results showed some significant changes (P<0.05) in the level of LDH leakage in normal cultured B50 cells (100%),hyoxic cells (440%) and treated cells with 100nM Anad (69%) and 100nM Arachi (103%). These changes in LDH release, neuronal viability, proliferation and differentiation was shown to be dose dependent between the normal and hypoxic B50 neurons and between treated and untreated B50 neurons in culture,while the RT-PCR mRNA levels showed no appreciable change. These results indicate that Cannabinoid agonists have some positive therapeutic effects in the treatment of hypoxia in neuronal B50 cells in culture.