Effects of Lewisite on cell membrane integrity and energy metabolism in human keratinocytes and SCL II cells.
Lewisite is a highly toxic arsenic compound which can cause skin damage. In the present study effects of Lewisite on cell membrane integrity and energy metabolism as well as antidotal effects of DL-2,3-dimercaptopropanesulfonate (DMPS), and meso-2,3-dimercaptosuccinic acid (m-DMSA) were investigated in a keratinocyte derived cell line (SCL II) and primary human keratinocytes (HK). Cells were incubated in Lewisite (60 microM) containing medium for 5 min. During the following 6 h lactate dehydrogenase (LDH) activity in the supernatant, intracellular ATP content, tetrazolium reduction, glucose consumption and lactate formation were measured. Glucose consumption and lactate production were decreased in both cell lines after Lewisite exposure. In SCL II cells an increase of LDH activity in the supernatant, a decrease of ATP content, and an impaired ability to reduce tetrazolium was found 3 h after Lewisite exposure. In HK cultures tetrazolium reduction was significantly decreased already after 2 h, whereas LDH increase in the supernatant and ATP content decrease occurred only at 6 h after Lewisite exposure. When DMPS or m-DMSA was added directly after Lewisite exposure to SCL II cells, glucose consumption and lactate formation were restored and LDH leakage was prevented. SCL II cells might be more prone to membrane damage whereas in keratinocytes mitochondrial impairment seems to be the predominant effect of Lewisite.
Kehe
K
K
Institute of Pharmacology and Toxicology, FAF Medical Academy, Neuherbergstr. 11, D-80937 Munich, Germany. kai.kehe@lrz.uni-muenchen.de
Flohé
S
S
Krebs
G
G
Kreppel
H
H
Reichl
F X
FX
Liebl
B
B
Szinicz
L
L
D000255:Adenosine Triphosphate / Q000032:analysis; D001152:Arsenicals / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents / Q000494:pharmacology; D004734:Energy Metabolism / Q000187:drug effects*; D005562:Formazans / Q000032:analysis; D005947:Glucose / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D019344:Lactic Acid / Q000032:analysis; D004113:Succimer / Q000494:pharmacology; D013778:Tetrazolium Salts / Q000032:analysis; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014494:Unithiol / Q000494:pharmacology
D001152:Arsenicals; D002614:Chelating Agents; D005562:Formazans; D013778:Tetrazolium Salts; C035965:lewisite; C079782:MTT formazan; D019344:Lactic Acid; D014494:Unithiol; D000255:Adenosine Triphosphate; D004113:Succimer; D007770:L-Lactate Dehydrogenase; D005947:Glucose