Effects of chloride channel inhibitors on H(2)O(2)-induced renal epithelial cell injury.
Oxidative stress contributes to renal epithelial cell injury in certain settings. Chloride influx has also been proposed as an important component of acute renal epithelial cell injury. The present studies examined the role of Cl(-) in H(2)O(2)-induced injury to LLC-PK(1) renal epithelial cells. Exposure of LLC-PK(1) cells to 1 mM H(2)O(2) resulted in the following: depletion of intracellular ATP content; DNA damage; lipid peroxidation; and a loss of membrane integrity to both small molecules, e.g., trypan blue, and macromolecules, e.g., lactate dehydrogenase (LDH), and cell death. Substitution of Cl(-) by isethionate or the inclusion of certain Cl(-) channel blockers, e.g., diphenylamine-2-carboxylate (DPC), 5-nitro-2-(3-phenylpropylamino). benzoate (NPPB), and niflumic acid, prevented the H(2)O(2)-induced loss of membrane integrity to LDH. In addition, the H(2)O(2)-induced loss of membrane integrity was prevented by raising the osmolality of the extracellular solutions, by depletion of cell ATP, and by inhibitors of volume-sensitive Cl(-) channels. However, these maneuvers did not prevent the H(2)O(2)-induced permeability to small molecules or H(2)O(2)-induced ATP depletion, DNA damage, lipid peroxidation, or cell death. These results support the view that volume-sensitive Cl(-) channels play a role in the progressive loss of cell membrane integrity during injury.
Meng
X
X
Division of Nephrology, University of Arkansas for Medical Sciences and John L. McClellan Memorial Veterans Hospital, Little Rock, Arkansas 72205, USA.
Reeves
W B
WB
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018118:Chloride Channels / Q000037:antagonists & inhibitors*; D002712:Chlorides / Q000378:metabolism; D004249:DNA Damage; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018374:LLC-PK1 Cells / Q000187:drug effects*; D015227:Lipid Peroxidation / Q000187:drug effects; D009579:Nitrobenzoates / Q000494:pharmacology; D018384:Oxidative Stress; D013552:Swine; D014343:Trypan Blue; D062367:ortho-Aminobenzoates / Q000494:pharmacology
D018118:Chloride Channels; D002712:Chlorides; D009579:Nitrobenzoates; D062367:ortho-Aminobenzoates; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D000255:Adenosine Triphosphate; C001209:fenamic acid; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D014343:Trypan Blue