| Title | Bepridil up-regulates cardiac Na+ channels as a long-term effect by blunting proteasome signals through inhibition of calmodulin activity. | | Author(s) | Kang L, Zheng MQ, Morishima M, Wang Y, Kaku T, Ono K | | Institution | Department of Pathophysiology, Oita University School of Medicine, Oita 879-5593, Japan. | | Source | Br J Pharmacol 2009 Jun; 157(3):404-14. | | MeSH | Animals
Animals, Newborn
Anti-Arrhythmia Agents
Bepridil
Calmodulin
Cells, Cultured
Dose-Response Relationship, Drug
Enzyme Activators
Humans
Muscle Proteins
Myocardium
Myocytes, Cardiac
Patch-Clamp Techniques
Proteasome Endopeptidase Complex
Protein Subunits
Rats
Rats, Wistar
Sodium Channels
Time Factors
Up-Regulation
| | Abstract | BACKGROUND AND PURPOSE: Bepridil is an anti-arrhythmic agent with anti-electrical remodelling effects that target many cardiac ion channels, including the voltage-gated Na+ channel. However, long-term effects of bepridil on the Na+ channel remain unclear. We explored the long-term effect of bepridil on the Na+ channel in isolated neonatal rat cardiomyocytes and in a heterologous expression system of human Na(v)1.5 channel. EXPERIMENTAL APPROACH: Na+ currents were recorded by whole-cell voltage-clamp technique. Na+ channel message and protein were evaluated by real-time RT-PCR and Western blot analysis. KEY
RESULTS: Treatment of cardiomyocytes with 10 micromol.L(-1) bepridil for 24 h augmented Na+ channel current (I(Na)) in a dose- and time-dependent manner. This long-term effect of bepridil was mimicked or masked by application of W-7, a calmodulin inhibitor, but not KN93 [2-[N-(2-hydroxyethyl)-N-(4-methoxy benzenesulphonyl)]-amino-N-(4-chlorocinnamyl)-N-methylbenzylamine], a Ca2+/calmodulin-dependent kinase inhibitor. During inhibition of protein synthesis by cycloheximide, the I(Na) increase due to bepridil was larger than the increase without cycloheximide. Bepridil and W-7 significantly slowed the time course of Na(v)1.5 protein degradation in neonatal cardiomyocytes, although the mRNA levels of Na(v)1.5 were not modified. Bepridil and W-7 did not increase I(Na) any further in the presence of the proteasome inhibitor MG132 [N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(1S)-1-formyl-3-methylbutyl]-L-leucinamide]. Bepridil, W-7 and MG132 but not KN93 significantly decreased 20S proteasome activity in a concentration-dependent manner.
CONCLUSIONS AND
IMPLICATIONS: We conclude that long-term exposure of cardiomyocytes to bepridil at therapeutic concentrations inhibits calmodulin action, which decreased degradation of the Na(v)1.5 alpha-subunit, which in turn increased Na+ current. | | Language | eng | | Pub Type(s) | Journal Article
Research Support, Non-U.S. Gov't
| | PubMed ID | 19371335 |
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