Research Article

Inhibition by Anandamide of 6-Hydroxydopamine-Induced Cell Death in PC12 Cells

Figure 1

Inhibition of 6-OHDA-induced apoptosis by anandamide. (a) PC12 cells were treated with 25  M anandamide for 24 hours and then exposed to 100  M 6-OHDA for a further 24 hours. Cytocentrifuge preparations of control and treated cells were stained with DAPI to visualise nuclear changes. The apoptotic cells with condensed and fragmented nuclei are indicated by arrows. Cells were (a) untreated, (b) exposed to 6-OHDA, (c) treated with AEA only or (d) treated with AEA and then exposed to 6-OHDA. (b) The level of cell death due to 6-OHDA with and without prior anandamide treatment was quantified and expressed as a percentage of the total number of cells. The results shown are the average of two separate experiments ± range. (c) PC12 cells were treated with a range of concentrations of anandamide (0–50  M) for 24 hours prior to exposure to 100  M 6-OHDA for a further 24 hours. DEVDase activity was measured in whole cell extracts. Values represent the mean ± SEM of four independent determinations. (d) PC12 cells were exposed to 25  M anandamide for 24 hours followed by treatment with 100  M 6-OHDA for further 24 hours. Pro-caspase-3 processing was visualized by Western blotting. Actin was used as a loading control. The data are representative of two independent experiments.