Effects of alkylxanthines on contractility of diaphragm fibres isolated from normal and sensitized guinea-pigs.

This study investigates the effects of alkylxanthines on twitch tension generated by electrical stimulation (supramaximal pulses, 0.2 ms duration, 1 Hz) of diaphragm muscle fibres isolated from normal and actively-sensitized guinea-pigs. Caffeine, theophylline and theobromine increased, in a concentration-dependent manner (50-500 microM), twitch tension in normal and sensitized diaphragm. Caffeine (500 microM) enhanced contractility to a greater extent than theophylline or theobromine. Twitch potentiation by caffeine (500 microM) was significantly greater in sensitized diaphragm. Verapamil (0.1-100 microM) did not alter twitch contractions in the absence or presence of alkylxanthines in normal or sensitized strips. Dantrolene (0.01-100 microM) depressed, in a concentration-dependent fashion, twitch contractions of normal and sensitized diaphragm. The inhibitory concentration 50% (expressed as -log IC50) was 6.78 +/- 0.13 in normal tissues and 6.15 +/- 0.11 in sensitized tissues (n = 6 in each group; P < 0.05). Exposure to Ca(2+)-free, EGTA (0.1 mM)-containing medium, depressed twitch contraction of normal diaphragm to a lesser extent than that of sensitized diaphragm. Methylxanthines reversed depression of twitch contractions produced by exposure to dantrolene (5 microM) or a Ca(2+)-free medium. Adenosine (1-1000 microM) was without effect whereas enprofylline (50-500 microM) enhanced diaphragm contractility in normal tissues. This indicates that blockade of adenosine receptors is not involved in the inotropic effect of alkylxanthines in guinea-pig diaphragm. Results from this study suggest that alkylxanthines enhance diaphragm contractility in the guinea-pig by releasing intracellular Ca2+ and promoting extracellular Ca2+ entry through verapamil-insensitive pathways. An alteration of Ca2+ movements and stores may be present in the sensitized diaphragm.