PT - JOURNAL ARTICLE AU - Rocchetti, Marcella AU - Besana, Alessandra AU - Mostacciuolo, Gaspare AU - Micheletti, Rosella AU - Ferrari, Patrizia AU - Sarkozi, Sandor AU - Szegedi, Csaba AU - Jona, Istvan AU - Zaza, Antonio TI - Modulation of Sarcoplasmic Reticulum Function by Na<sup>+</sup>/K<sup>+</sup> Pump Inhibitors with Different Toxicity: Digoxin and PST2744 [(<em>E</em>,<em>Z</em>)-3-((2-Aminoethoxy)imino)androstane-6,17-dione Hydrochloride] AID - 10.1124/jpet.104.077933 DP - 2005 Apr 01 TA - Journal of Pharmacology and Experimental Therapeutics PG - 207--215 VI - 313 IP - 1 4099 - http://jpet.aspetjournals.org/content/313/1/207.short 4100 - http://jpet.aspetjournals.org/content/313/1/207.full SO - J Pharmacol Exp Ther2005 Apr 01; 313 AB - Objective: To gain some insight on the lesser arrhythmogenic properties of PST2744 [(E,Z)-3-((2-aminoethoxy)imino)androstane-6,17-dione hydrochloride] compared with digoxin, we compared modulation of intracellular Ca2+ dynamics by the two agents. Methods: SERCA (sarcoplasmic reticulum Ca2+-ATPase) activity and Ca2+ leak rate were measured in sarcoplasmic reticulum (SR) vesicles from guinea pig ventricles. Membrane current, intracellular Ca2+, and twitch amplitude were evaluated in guinea pig ventricular myocytes with or without blockade of the Na+/Ca2+ exchanger. Results: In SR vesicles, PST2744 (30–300 nM), but not digoxin, increased SERCA activity; digoxin only (≥0.1 nM) increased SR Ca2+ leak. In myocytes with blocked Na+/Ca2+ exchanger, Ca2+ reloading of caffeine-depleted SR was enhanced by PST2744 and slightly inhibited by digoxin. In myocytes with functioning Na+/Ca2+ exchanger, both agents increased diastolic Ca2+, SR Ca2+ content, the gain of Ca2+-induced Ca2+ release, the rate of cytosolic Ca2+ decay, twitch amplitude, and relaxation rate. Consistent with the observations in SR vesicles, the effects on SR Ca2+ content and Ca2+ decay rate were significantly larger for PST2744 than for digoxin. Conclusions: In isolated SR vesicles, PST2744 and digoxin directly affected SR function in opposite ways; this could be reproduced in myocytes during Na+/Ca2+ exchanger blockade. Under physiological conditions (functioning Na+/Ca2+ exchanger), the two agents affected Ca2+ dynamics in the same direction, as expected by their Na+/K+ pump inhibition; however, differential SR modulation was still expressed by quantitative differences. Thus, the more favorable inotropy-to-toxicity ratio previously described for PST2744 appears to be associated with direct SERCA stimulation and/or lack of enhancement of Ca2+ leak. The American Society for Pharmacology and Experimental Therapeutics