Pre-Conditioning with CDP-Choline Attenuates Oxidative Stress-Induced Cardiac Myocyte Death in a Hypoxia/Reperfusion Model

Figure 3

CDP-choline attenuates hypoxia/reperfusion-induced apoptosis. Cardiac myocyte cultures were subjected to coverslip hypoxia/reperfusion alone (Hypoxia/reperfusion), incubated with CDP-choline (50 mg/mL) prior to reperfusion (CDP-choline preconditioning), or incubated with CDP-choline (50 mg/mL) 1 hr after reperfusion (CDP-choline postconditioning). Cell cultures were left untreated as a control (Negative control). After treatment, the cells were stained with hematoxylin-eosin; cell morphology was assessed by light microscopy (40X) (upper panel). Apoptosis and necrosis were evaluated by flow cytometry analysis of cells labeled with Annexin-V-FLUOS (Annexin V) and propidium iodide (PI) (lower panel). The proportion of labeled cells is depicted in the lower panel. The proportion of viable cells, with low Annexin and low PI staining, is shown in the left, lower quadrant of each dot plot. Apoptotic cells, with high Annexin and low PI staining, are depicted in the right, lower quadrant. Necrotic cells, with high Annexin and high PI staining, are showed in the right, upper quadrant. Results shown are representative figures from three independent assays.