Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2013 Apr 1.

Published in final edited form as: Kidney Int. 2012 Jun 13;82(8):878–891. doi: 10.1038/ki.2012.224

A. Lactate dehydrogenase (LDH) release as a marker of hydrogen peroxide (H₂O₂)-induced necrosis in HK-2 cells (N=6 for each group, expressed as a percentage of total LDH released). Vehicle (0.00003% DMSO) or CCPA (1 μM) were applied 6 hr before H₂O₂ treatment, and a selective SK1 inhibitor (SKI-II, 10 μM) or a selective S1P₁R inhibitor (W146, 1 μM) was given 30 min before vehicle or CCPA. A₁AR activation significantly attenuated H₂O₂-induced necrosis in HK-2 cells and this protective effect was blocked by the inhibition of SK1 or S1P₁R signaling. Data are presented as means ± SEM. #P<0.05 vs. vehicle treatment. B. Representative poly(adenosine diphosphate-ribose) polymerase (PARP) and Caspase-3 fragmentation (N=4 for each group) as indices of HK-2 cell apoptosis. HK-2 cell apoptosis was induced by TNF-α (20 ng/mL) and cycloheximide (10 μg/mL), and vehicle (0.00003% DMSO) or CCPA (1 μM) were applied 30 min before induction of apoptosis. A selective SK1 inhibitor (SKI-II, 1 μM given 30 min before vehicle or CCPA treatment) reduced the anti-apoptotic effect of A₁AR activation in HK-2 cells. Representative of 4 independent experiments.

A. Lactate dehydrogenase (LDH) release as a marker of hydrogen peroxide (H₂O₂)-induced necrosis in HK-2 cells (N=6 for each group, expressed as a percentage of total LDH released). Vehicle (0.00003% DMSO) or CCPA (1 μM) were applied 6 hr before H₂O₂ treatment, and a selective SK1 inhibitor (SKI-II, 10 μM) or a selective S1P₁R inhibitor (W146, 1 μM) was given 30 min before vehicle or CCPA. A₁AR activation significantly attenuated H₂O₂-induced necrosis in HK-2 cells and this protective effect was blocked by the inhibition of SK1 or S1P₁R signaling. Data are presented as means ± SEM. #P<0.05 vs. vehicle treatment. B. Representative poly(adenosine diphosphate-ribose) polymerase (PARP) and Caspase-3 fragmentation (N=4 for each group) as indices of HK-2 cell apoptosis. HK-2 cell apoptosis was induced by TNF-α (20 ng/mL) and cycloheximide (10 μg/mL), and vehicle (0.00003% DMSO) or CCPA (1 μM) were applied 30 min before induction of apoptosis. A selective SK1 inhibitor (SKI-II, 1 μM given 30 min before vehicle or CCPA treatment) reduced the anti-apoptotic effect of A₁AR activation in HK-2 cells. Representative of 4 independent experiments.