Left regional cardiac perfusion in vitro with platelet-activating factor, norepinephrine and K+ reveals that ischaemic arrhythmias are caused by independent effects of endogenous "mediators" facilitated by interactions, and moderated by paradoxical antagonism

Abstract

Various putative drug targets for suppression of ischaemia-induced ventricular fibrillation (VF) have been proposed, but therapeutic success in the suppression of sudden cardiac death (SCD) has been disappointing. Platelet-activating factor (PAF) is a known component of the ischaemic milieu. We examined its arrhythmogenic activity, and its interaction with two other putative mediators, norepinephrine and K(+), using an ischaemia-free in vitro heart bioassay, and a specific PAF antagonist (BN-50739). PAF (0.1-100 nmol) was administered selectively to the left coronary bed of rat isolated hearts using a specially designed catheter. In some hearts, PAF was administered to the left coronary bed during concomitant regional perfusion with norepinephrine and/or K(+). In separate studies, PAF accumulation in the perfused cardiac tissue was evaluated using (3)H-PAF. PAF evoked ventricular arrhythmias concentration-dependently (P<0.05). It also widened QT interval and reduced coronary flow selectively in the PAF-exposed left coronary bed (both P<0.05). Two exposures of hearts to PAF were necessary to evoke the QT and rhythm effects. The PAF-induced arrhythmias and coronary vasoconstriction were partially suppressed by the PAF antagonist BN-50739 (10 microm), although BN-50739 itself widened QT interval. K(+) (8 and 15 mm) unexpectedly antagonised the arrhythmogenic effects of PAF without itself eliciting arrhythmias (P<0.05). Norepinephrine (0.1 microm) had little or no effect on the actions of PAF, while failing to evoke arrhythmias itself. Nevertheless, the combination of 15 mm K(+) and 0.1 microm norepinephrine evoked arrhythmias of a severity similar to arrhythmias evoked by PAF alone, without adding to or diminishing the arrhythmogenic effects of PAF. (3)H-PAF accumulated in the cardiac tissue, with 43+/-5% still present 5 min after bolus administration, accounting for the need for two exposures of the heart to PAF for evocation of arrhythmias. Thus, PAF, by activating specific receptors in the ventricle, can be expected to contribute to arrhythmogenesis during ischaemia. However, its interaction with other components of the ischaemic milieu is complex, and selective block of its actions (or its accumulation) in the ischaemic milieu is alone unlikely to reduce VF/SCD.

Figure 1

First experimental protocol used to investigate the actions of exogenous PAF in the absence of ischaemia. The left coronary bed was perfused with Krebs, then exposed to PAF or vehicle by bolus injection into the perfusion line at the concentrations and intervals indicated.

Figure 2

Second protocol in which the left coronary bed was exposed to PAF, then PAF or vehicle during continuous perfusion with K⁺ and/or norepinephrine.

Figure 3

Third protocol, in which the left coronary bed was perfused with BN-50739 or its vehicle (DMSO) during exposure to bolus injections of PAF.

Figure 4

Change in coronary flow in the left and right coronary beds during exposure to bolus injection of PAF (without prior exposure) into the left coronary perfusion line. ^*P<0.05 versus values for the right coronary bed.

Figure 5

QT interval in the left coronary bed during a second exposure to PAF by bolus injection into the left coronary perfusion line. ^*P<0.05 versus 0 nmol PAF.