Apoptosis at a distance: Remote activation of caspase-3 occurs early after myocardial infarction

Objective: After an acute myocardial infarction, the viable myocardium remote from the infarct zone is subjected to ventricular remodeling. Besides hypertrophy, processes of apoptosis may contribute to these remodeling processes. Reports on apoptosis in this area have been doubted because they were mainly based on in-situ nick-end DNA labeling (TUNEL) measurements, with questionable specifity. Moreover, the time course of initiation of these processes has not been characterized. Therefore the goals of this study were to (1) reliably determine if in the remote area of the infarcted heart apoptosis may be initiated using highly specific biochemical markers and (2) evaluate the time course of such an activation. Methods: A well-defined model, regional myocardial infarction induced by ligation of the left anterior coronary artery in rats in vivo, was used. Heart and lung wet weights, the left ventricular end-diastolic pressure (LVEDP), and the serum level of the atrial natriuretic propeptide (proANP) were determined from 1 day up to 4 weeks as indicators of developing heart failure. In transmural biopsies from the non-ischemic left ventricular wall of the infarcted heart, the activation of caspase-3, the bcl-2/bax ratio (Western blot analysis), and the DNA laddering (LM-PCR) were determined. Results: Although heart- and lung weights did not increase before 1 week after infarction, proANP levels were elevated already 1 day after myocardial infarction suggesting early sub-clinical heart failure. The activity of caspase-3 increased significantly to 160 +/- 20% compared to sham operated controls as early as 1 day after ligation and remained elevated over the entire time course. In parallel, the bcl-2/bax ratio shifted toward the pro-apoptotic bax. Moreover, these clear and specific biochemical indicators of apoptosis in the remote area of the infarcted heart were paralleled by the fragmentation of genomic DNA. Conclusion: These data demonstrate that apoptotic markers are activated in the surviving zone of the heart remote from the infarct area as early as 1 day after myocardial infarction with persistence for up to 4 weeks. This activation coincides with early markers of heart failure. The exact regulation of this apoptotic process remains to be elucidated.