Oxidative modulation of marcaine and lekoptin in H9C2 rat myoblasts.
OBJECTIVE
The cytotoxicity of marcaine was estimated in combination with a calcium channel blocker. In addition, the influence of marcaine and marcaine plus lekoptin on a model system using the H9C2 cardiac cell line was investigated.


METHODS
Cells were incubated for five hours with marcaine, lekoptin, or with both drugs simultaneously. Apoptotic cells were detected using the TUNEL assay and the alkaline comet assay. Mitochondrial cell function after drug uptake was examined using the MTT assay. The concentration of MDA (malondialdehyde) -- the final product of fatty-acid peroxidation, was quantified spectrophotometrically. The expression of glutathione S-transferase pi (GST-pi) was detected by immunofluorescence (IF) and Western blotting (WB) and inducible nitric oxide synthase (iNOS) was assessed by immunocytochemical staining (ABC).


RESULTS
Incubation with marcaine resulted in the highest number of apoptotic cells. After incubation with both marcaine and lekoptin, moderate damage to cells (54.2%+/-1.775% of DNA destruction) was observed. The highest levels of iNOS and GST-pi expression were observed in cells treated with marcaine and marcaine plus lekoptin. The characteristic nuclear GST-pi expression was observed in cells treated with both drugs.


CONCLUSIONS
Lekoptin stimulated cells to proliferate. Marcaine caused membrane damage and ultimately cell death.
Kulbacka
Julita
J
Department of Medical Biochemistry, Wroclaw Medical University, Wroclaw, Poland. julitab@bioch.am.wroc.pl
Bar
Julita
J
Chwilkowska
Agnieszka
A
Dumanska
Malgorzata
M
Drag-Zalesinska
Malgorzata
M
Wysocka
Teresa
T
Stach
Kamilla
K
Bednarz
Iwona
I
Lugowski
Mateusz
M
Marcinkowska
Anna
A
Gamian
Andrzej
A
Saczko
Jolanta
J
D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002045:Bupivacaine / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D002460:Cell Line; D019394:Ki-67 Antigen / Q000378:metabolism; D015227:Lipid Peroxidation; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D014700:Verapamil / Q000494:pharmacology*
D000779:Anesthetics, Local; D002121:Calcium Channel Blockers; D019394:Ki-67 Antigen; D014700:Verapamil; D052247:Nitric Oxide Synthase Type II; D002045:Bupivacaine