Oxidative modulation of marcaine and lekoptin in H9C2 rat myoblasts

doi:doi:10.1038/aps.2008.27

Acta Pharmacologica Sinica (2009) 30: 184–192; doi: 10.1038/aps.2008.27

Oxidative modulation of marcaine and lekoptin in H9C2 rat myoblasts

Julita Kulbacka¹, Julita Bar³, Agnieszka Chwilkowska¹, Malgorzata Dumanska², Malgorzata Drag-Zalesinska², Teresa Wysocka², Kamilla Stach¹, Iwona Bednarz¹, Mateusz Lugowski¹, Anna Marcinkowska¹, Andrzej Gamian¹ and Jolanta Saczko¹

¹Department of Medical Biochemistry, Wroclaw Medical University, 50-368 Wroclaw, Poland
²Department of Histology and Embryology, Wroclaw Medical University, 50-368 Wroclaw, Poland
³Department of Clinical Immunology, Medical University, 50-368 Wroclaw, Poland

Correspondence: Dr Julita Kulbacka, PhD., E-mail julitab@bioch.am.wroc.pl

Received 24 September 2008; Accepted 18 December 2008.

Top

Abstract

Aim:

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 (GST-) 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% plusminus 1.775% of DNA destruction) was observed. The highest levels of iNOS and GST- expression were observed in cells treated with marcaine and marcaine plus lekoptin. The characteristic nuclear GST- expression was observed in cells treated with both drugs.