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Division of Nephrology, University of Arkansas for Medical Sciences and John L. McClellan Memorial Veterans Hospital, Little Rock, Arkansas 72205
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 H2O2-induced
injury to LLC-PK1 renal epithelial cells. Exposure of
LLC-PK1 cells to 1 mM H2O2 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 H2O2-induced loss of
membrane integrity to LDH. In addition, the
H2O2-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 H2O2-induced permeability to small
molecules or H2O2-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.
oxidant stress; chloride channels; LLC-PK1 cell; necrosis
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