Abstract
Purpose
This study examined the direct myocardial depressant effect of halothane and determined whether an L-type Ca2+ channel agonist and antagonists altered the myocardial depression induced by halothane in cultured rat ventricular myocytes.
Methods
Ventricular myocytes were obtained from neonatal rats by enzymatic digestion with collagenase and then cultured for 6 to 7 days. The myocytes were stabilized in a serum-free medium, and the spontaneous beating rate and amplitude were measured. To assess the halothane-induced conformational changes in L-type Ca2+ channel, receptor binding study was performed using a dihydropyridine derivative, [3H] PN 200-110, in cardiac membrane preparation.
Results
Halothane (1%, 2%, 3%, 4%) decreased the beating rate and amplitude in a concentration-dependent manner (P < 0.05). The myocardial depressant effects of halothane were potentiated by nifedipine or verapamil (P < 0.05). Bay K 8644, an L-type Ca2+ channel agonist, completely prevented the halothane-induced depression in amplitude (P < 0.05), but affected the beating rate less. Adding halothane (2%) decreased (P < 0.05) the maximum binding site density for [3H] PN 200-110 (from 198.6 ± 23.7 fmol·mg−1 protein to 115.3 ± 21.6 fmol·mg−1 protein) but did not affect binding affinity (from 0.461 ± 0.077 nM to 0.307 ± 0.055 nM).
Conclusion
The reduction of Ca2+ current via sarcolemmal L-type Ca2+ channel, probably due to conformational changes in dihydropyridine binding sites, plays an important role in halothane-induced myocardial depression in living heart cells.
Résumé
Objectif
Létude actuelle a examiné l’effet dépresseur myocardique direct de l’halothane et déterminé si un agoniste ou un antagoniste des canaux calciques de type L modifie la dépression myocardique induite par l’halothane sur des myocytes ventriculaires de rats, en culture.
Méthodes
Les myocytes ventriculaires ont été obtenus à partir de rats nouveau-nés, par digestion enzymatique avec collagénase, et puis mis en culture pendant 6 ou 7 jours. Les myocytes ont été stabilisés dans un milieu sans sérum et le rythme et l’amplitude des battements spontanés ont été mesurés. Pour évaluer les changements de conformation induits par l’halothane dans le canal calcique de type L, l’étude de la liaison aux récepteurs a été réalisée en utilisant un dérivé dihydropyridine, [3H] PN 200-110, d’une préparation de membranes cardiaques.
Résultats
L’halothane (1 %, 2 %, 3 %, 4%) a diminué le rythme des battements et leur amplitude en fonction de la concentration (P < 0,05). Les effets dépresseurs myocardiques de l’halothane ont été accentués par la nifédipine ou le vérapamil. Le Bay K 8644, un agoniste des canaux calciques de type L, a complètement empêché la dépression d’amplitude induite par l’halothane (P < 0,05), mais a eu moins d’effet sur le rythme des battements. L’accroissement de l’halothane (2 %) a diminué (P < 0,05) la densité maximale au site de liaison pour le [3H] PN 200-110 (de 198,6 ± 23,7 fmol·mg−1 de protéine à 115,3 ± 21,6 fmol·mg−1 de protéine), mais n’a pas affecté l’affinité de la liaison (de 0,461 ± 0,077 nM à 0,307 ± 0,055 nM).
Conclusion
La réduction de l’entrée de calcium dans le canal calcique sarcolemmique de type L joue un rôle important dans la dépression myocardique induite par l’halothane dans des cellules cardiaques en culture à cause, probablement, des changements de conformation des sites de liaison de la dihydropyridine.
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Supported by Grants-in-Aid for scientific research (No. 06771227, 06671545 and 06670121) from the Ministry of Education, Science and Culture, Japan.
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Kanaya, N., Matsumoto, M., Kawana, S. et al. Ca2+ channel modulation alters halothane-induced depression of ventricular myocytes. Can J Anaesth 45, 584–591 (1998). https://doi.org/10.1007/BF03012714
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DOI: https://doi.org/10.1007/BF03012714