Figure 2 | Cell Death & Disease

Figure 2

From: Cannabidiol protects oligodendrocyte progenitor cells from inflammation-induced apoptosis by attenuating endoplasmic reticulum stress

Figure 2

CBD protects against inflammatory damage by diminishing apoptosis and decreases the number of TUNEL+ OPCs through a mechanism that does not involve CB1, CB2, TRPV1 or PPARγ receptors. (a) LPS/IFNγ-induced cytotoxicity in OPCs was attenuated by CBD, a dose of 1 μM proving to be the most effective). Administration of CB1, CB2, TRPV1 or PPARγ antagonists (SR1, AM639, CPZ: 1 μM and GW9662: 50 nM) 30 min before the stimulus had no effect, indicating that none of these receptors are implicated in the protective effects of CBD. The data represent the mean±S.E.M. of n=3 independent cultures analyzed in triplicate, and the statistical analysis was performed using Kruskal–Wallis ANOVA followed by Mann–Whitney U test: ***P⩽0.001 versus untreated cells, #P⩽0.05 and ###P⩽0.001 versus cells exposed to LPS/IFNγ alone. (b) Cleaved caspase3 western blot shows that LPS/IFNγ treatment induces the activation of the apoptotic pathway, whereas the co-treatment with CBD reduces this induction. OPCs were incubated with LPS/IFNγ in presence or absence of CBD (1 μM). Total protein extracts were prepared 24 h later and cleaved caspase 3 (19 KDa) was assessed in western blots probed with specific antibodies. The data represent the mean±S.E.M. optical density normalized to tubulin from three independent cultures analyzed in triplicate, and statistical analysis was performed using one-way ANOVA followed by the Bonferroni post-hoc test: **P⩽0.01 compared with non-treated cells, #P⩽0.05 compared with LPS/IFNγ group. (c and d) A2B5 and TUNEL staining show that LPS/IFNγ decreased OPC number by inducing apoptosis, an effect that was reversed by CBD as previously confirmed by caspase 3 measurement. OPCs were exposed for 24 h to the cytotoxic stimulus, in the presence or absence of CBD (1 μM), and 6000 OPCs cells were counted. Data represent the mean±S.E.M., and the statistical significance was determined using the Kruskal–Wallis ANOVA followed by Mann–Whitney U test: ***P⩽0.001 versus untreated cells, and ###P⩽0.001 versus cells exposed to LPS/IFNγ alone

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