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Delta-9-Tetrahydrocannabinol and other phytocannabinoids have been previouslydemonstrated to possess neuroprotective effects in murine mesencephalic cellculture models of Parkinson’s disease, in which increased levels ofsuperoxide radicals led to the loss of dopaminergic neurons. In these models,delta-9-tetrahydrocannabinol did not scavenge these radicals but displayedantioxidative capacity by increasing glutathione levels. Based on thesefindings, in the present study, we investigated whether the neuroprotectiveeffect of delta-9-tetrahydrocannabinol can also be detected in FeSO4-and H2O2-stressed cells. Mesencephalic cultures wereconcomitantly treated with FeSO4 (350 μM) orH2O2 (150 μM) anddelta-9-tetrahydrocannabinol (0.01, 0.1, 1, 10 μM) on the12thdays in vitro for 48 h. On the 14th DIV, dopaminergicneurons were stained immunocytochemically by tyrosine hydroxylase, andfluorescently using crystal violet, Hoechst 33342, and JC-1. FeSO4and H2O2 significantly reduced the number of dopaminergicneurons by 33 and 36%, respectively, and adversely affected themorphology of surviving neurons. Moreover, FeSO4, but notH2O2, significantly decreased the fluorescenceintensity of crystal violet and Hoechst 33342, and reduced the red/greenratio of JC-1. Co-treatment with delta-9-tetrahydrocannabinol at theconcentrations 0.01 and 0.1 μM significantly rescueddopaminergic neurons in FeSO4 and H2O2-treatedcultures by 16 and 30%, respectively. delta-9-Tetrahydrocannabinoltreatment also led to a higher fluorescence intensity of crystal violet andHoechst 33342, and increased the red/green fluorescence ratio of JC-1when concomitantly administered with FeSO4 but notH2O2. To conclude, delta-9-tetrahydrocannabinolrescues dopaminergic neurons against FeSO4- andH2O2-induced neurotoxicity. Using fluorescence dyes,this effect seems to be mediated partially by restoring mitochondrial integrityand decreasing cell death, particularly in FeSO4-treatedcultures.

planta medica international open

Neuroprotective Effects of Delta-9-Tetrahydrocannabinol against FeSO4- and H2O2-Induced Cell Damage on Dopaminergic Neurons in Primary Mesencephalic Cell Culture