Toxicity to isolated hepatocytes caused by the intracellular calcium indicator, Quin 2.
To determine whether incubation for several hours with intracellular Ca++ indicators caused toxicity to freshly isolated hepatocytes from rats, cells were incubated under 95% O2-5% CO2 in medium containing 2 mM Ca++ and the acetoxymethyl (AM) esters of Quin 2, Indo 1, Fluo 3, 5,5'-Dimethyl BAPTA, 4,4'-Difluoro BAPTA or Fura 2 for up to 5 hr. Quin 2-AM and Indo 1-AM (2.5 microM) induced lipid peroxidation in the cells after 1 or 3 hr of treatment, respectively. Additional experiments with Quin 2-AM (25 microM) revealed that it also caused lactate dehydrogenase leakage, cell blebbing and vitamin E loss in cells, but did not affect reduced glutathione or intracellular Ca++ content. The ability of Quin 2-AM to cause toxicity was dependent on the amount of Quin 2 which was present in the cell. Ca++ appeared to be involved in the mechanism of Quin 2-AM toxicity, for modulation of the extracellular Ca++ concentration partially inhibited lipid peroxidation, vitamin E loss, cell blebbing and lactate dehydrogenase leakage.
Carpenter-Deyo
L
L
Department of Biochemistry and Biophysics, Oregon State University, Corvallis.
Duimstra
J R
JR
Hedstrom
O
O
Reed
D J
DJ
D000634:Aminoquinolines / Q000633:toxicity*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002614:Chelating Agents / Q000633:toxicity*; D005456:Fluorescent Dyes / Q000633:toxicity*; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D014810:Vitamin E / Q000378:metabolism
D000634:Aminoquinolines; D002614:Chelating Agents; D005456:Fluorescent Dyes; D014810:Vitamin E; C035738:Quin2-acetoxymethyl ester; D007770:L-Lactate Dehydrogenase; D002118:Calcium