Figure 3.

Tumor necrosis factor-α plus cholecystokinin induce apoptosis and necrosis in pancreatic acini. A-E: Isolated pancreatic acini were stimulated with either 10 ng/mL tumor necrosis factor-α (TNF-α) for 2 h, or 50 pmol/L cholecystokinin (CCK) for 1 h, or 10 ng/mL TNF-α for 1 h followed by 10 ng/mL TNF-α plus 50 pmol/L CCK for 1 h, or vehicle-control for 2 h. A: Lactate dehydrogenase (LDH) was measured by the DGKC optimized kinetic method using the respective assay kit. Results are expressed as a percentage of total cellular LDH determined by permeabilizing cells with Triton X-100, and correspond to the mean ± SE from four independent experiments, with samples performed in triplicate. ^aP < 0.05 vs vehicle-control; B: Caspases 2, 3, 8, and 9 activity was measured by a colorimetric/fluorometric method using the respective assay kits. Results are expressed as fold of control and correspond to the mean ± SE from four independent experiments, with samples performed in triplicate. ^aP < 0.05, ^bP < 0.01 vs vehicle-control; C-E: ATP and ADP levels were measured by a bioluminescent method using the respective assay kits. ATP loss was calculated as a percentage of the decrease in values corresponding to vehicle-control. Results correspond to the mean ± SE from four independent experiments, with samples performed in triplicate. ^aP < 0.05 vs vehicle-control.