Bupivacaine-induced Apoptosis in the Primary Cultured Cardiomyocytes via p38 MAPKs.
10.4097/kjae.2006.50.6.S48
- Author:
Hyun Jeong KIM
1
;
Se Ra SUNG
;
Kwang Suk SEO
;
Seung Woon LIM
;
Tae Gyoon YOON
Author Information
1. Department of Dental Anesthesiology, Seoul National University College of Dentistry, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
apoptosis;
bupivacaine;
calcium;
cardiomyocyte;
levobupivacaine;
mitogen-activated protein kinases
- MeSH:
Anesthetics, Local;
Animals;
Apoptosis*;
Blotting, Western;
Bupivacaine;
Calcium;
Caspase 3;
Cell Death;
Chelating Agents;
Chromatin;
Cytochromes c;
Cytoplasm;
DNA Fragmentation;
Electrophoresis;
Extracellular Signal-Regulated MAP Kinases;
Humans;
Mitogen-Activated Protein Kinases;
Myocytes, Cardiac*;
p38 Mitogen-Activated Protein Kinases*;
Phosphotransferases;
Protein Kinases;
Rats
- From:Korean Journal of Anesthesiology
2006;50(6):S48-S56
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: It is known that bupivacaine induce cell death in several immortalized cells. However, there is no report concerning bupivacaine-induced cell death in the primary cultured cardiomyocytes. We compared the direct cytotoxicity of local anesthetics in cardiomyocytes. Furthermore, the mechanisms of cell death were evaluated. METHODS: The myocardial cells of rat pups were cultured 3 days after seeding. The methyltetrazolium (MTT) assay was employed to quantify differences in cellular viability. To confirm apoptosis, Hoechst-propidium iodide staining, DNA fragmentation by electrophoresis and western blot analysis were performed. And to examine the mechanisms of cell death, intracellular calcium and expression levels of mitogen-activated protein kinases (MAPKs) family members were evaluated. RESULTS: Among the local anesthetics under 1 mM concentration for 18 h, only bupivacaine significantly decreased the MTT activity (P < 0.001). Bupivacaine induced cell death in a dose-responsive and time dependent manner. Cell death showed apoptotic characteristics, such as DNA fragmentation, chromatin condensation, decrease of precursor caspase-3 protein level, increased cleaved PARP, and cytochrome C release into the cytoplasm. Bupivacaine phosphorylated three major MAPKs, i.e. extracellular signal-regulated kinases (ERKs), p38 kinase and c-Jun N-terminal kinases (JNKs) stress-activated protein kinases. Administration of ERK inhibitor increase cell death, whereas inhibitors of p38 kinase and JNK decreased cell death (P < 0.05). In addition, the intracellular calcium level was approximately 4 times higher after the bupivacaine treatment (P < 0.001), which was inhibited by calcium chelators (P < 0.001). Calcium chelators inhibited expression of MAPKs. CONCLUSIONS: In bupivacaine-induced apoptosis in cardiomyocytes, intracellular calcium increase and MAPKs family plays important roles.
