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Title[redir]:
Hepatology
Description:
ovement of chelatable iron in oxidative stress-induced killing to cultured hepatocytes from C3Heb mice and Sprague-Dawley rats. Mitochondrial polarization and chelatable iron were visualized by confocal microscopy of tetramethylrhodamine methylester (TMRM) and quenching of calcein, respectively. Cell viability and hydroperoxide formation (a measure of lipid peroxidation) were measured fluorometrically using propidium iodide and chloromethyl dihydrodichlorofluorescein, respectively. After collapse of lysosomal/endosomal acidic pH gradients with bafilomycin (50 nM), an inhibitor of the vacuolar proton-pumping adenosine triphosphatase, cytosolic calcein fluorescence became quenched. Deferoxamine mesylate and starch-deferoxamine (1 mM) prevented bafilomycin-induced calcein quenching, indicating that bafilomycin induced release of chelatable iron from lysosomes/endosomes. Bafilomycin also quenched calcein fluorescence in mitochondria, which was blocked by 20 μM Ru360, an inhibitor of the mitochondrial calcium uniporter, consistent with mitochondrial iron uptake by the uniporter. Bafilomycin alone was not sufficient to induce mitochondrial depolarization and cell killing, but in the presence of low-dosetert-butylhydroperoxide (25 μM), bafilomycin enhanced hydroperoxide generation, leading to mitochondrial depolarization and subsequent cell death.Conclusion:Taken together, the results are consistent with the conclusion that bafilomycin induces release of chelatable iron from lysosomes/endosomes, which is taken up by mitochondria. Oxidative stress and chelatable iron thus act as two “hits” synergistically promoting toxic radical formation, mitochondrial dysfunction, and cell death. This pathway of intracellular iron translocation is a potential therapeutic target against oxidative stress–mediated hepatotoxicity. (Hepatology 2008.) Abbreviations: cmDCF, chloromethyldichlorofluorescein; cmH2DCF-DA, chloromethyldihydrodichlorofluorescein diacetate; HEPES, 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid; MPT, mitochondrial permeability transition; O2•−, superoxide; OH•, hydroxyl radical; PI, propidium iodide; ROS, reactive oxygen species; t-BuOOH, tert-butylhydroperoxide; TMRM, tetramethylrhodamine methylester....
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Keywords {🔍}
iron, liver, mitochondrial, bafilomycin, aasld, chelatable, hepatology, log, journal, oxidative, calcein, cell, access, subscribe, issue, alerts, abstract, mitochondria, diseases, register, member, logo, browse, submit, article, translocation, lysosomes, stressinduced, hepatocellular, injury, buy, hepatocytes, role, intracellular, killing, mice, tetramethylrhodamine, methylester, tmrm, quenching, hydroperoxide, formation, propidium, iodide, inhibitor, fluorescence, quenched, release, lysosomesendosomes, uniporter,
Topics {✒️}
acetaminophen–induced hepatotoxicity back oxidative stress–mediated hepatotoxicity oxidative stress-induced killing aasld members log cholestatic liver open access policy individual subscribers log bafilomycin induced release lysosomal iron participates institutional users access sprague-dawley rats cytosolic calcein fluorescence potential therapeutic target 1-piperazine ethanesulfonic acid reactive oxygen species bile acid homeostasis artificial intelligence training intracellular iron translocation bafilomycin induces release mitochondrial permeability transition low-dosetert-butylhydroperoxide liver diseases full text access induce mitochondrial depolarization mitochondrial iron uptake quenched calcein fluorescence oxidative stress subsequent cell death mitochondrial calcium uniporter hydroperoxide formation journal tables register subscribe hepatology 2008 service request 800-638-3030 hydroxyl radical cell killing mitochondrial depolarization journal cell death website subscribe mitochondrial polarization mitochondrial dysfunction intracellular movement tert-butylhydroperoxide heme iron chelatable iron cell viability key event jae-sung3 precise mechanisms
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