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Am J Physiol Heart Circ Physiol (July 1, 2004). doi:10.1152/ajpheart.01143.2003
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Submitted on December 4, 2003
Accepted on June 1, 2004

Ischemic preconditioning and morphine attenuate myocardial apoptosis and infarction following ischemia/reperfusion in Rabbits- Role of {delta}-opioid receptor

Shinji Okubo1*, Yujirou Tanabe1, Kenji Takeda1, Michihiko Kitayama1, Seiyu Kanemitsu1, Rakesh C. Kukreja2, and Noboru Takekoshi1

1 Departmentof Cardiology, Kanazawa Medical University, Kanazawa, Uchinada, Japan
2 Division of Cardiology, Department of Medicine, Virginia Commonwealth University Medical Center, Richmond, Virginia, USA

* To whom correspondence should be addressed. E-mail: shinkubo{at}kanazawa-med.ac.jp.

We examined whether ischemic preconditioning (IPC) attenuates ischemia/reperfusion injury in part by decreasing apoptosis and if the {delta}-opioid receptor (DOR) plays a pivotal role in the regulation of apoptosis. Rabbits were subjected to 30-min coronary artery occlusion (CAO) and 180-minutes of reperfusion. IPC was elicited with 4 cycles of 5-min ischemia and 10-min reperfusion before CAO. Morphine (0.3 mg/kg iv) was given 15 min before CAO. Naloxone (Nal, 10 mg/kg, iv) and naltrindole (NTI, 10 mg/kg), the respective non-selective and selective {delta}-opioid receptor antagonists were given 10 min prior to either morphine or IPC. Infarct size (% risk area) was reduced from 46±3.8 in control to 11.6 ± 1.0 in IPC and 19.5 ± 3.8 in morphine group (mean±SEM, P<0.001 vs control). Naloxone blocked protective effects of IPC and morphine as shown by increase in infarct size to 38.6±7.2 and 44.5±1.8 respectively. Similarly, NTI blocked IPC and morphine-induced protection. Percentage of apoptotic cells (revealed by TUNEL assay) decreased in IPC (3.6±1.9) and morphine groups (5.2±1.2) as compared to control group (12.4±1.6, P<0.001). NTI pretreatment increased apoptotic cells 11.2±2.2% in IPC and 12.1±0.8% in morphine groups. Nal failed to block inhibition of apoptosis in the IPC group (% of cells 5.7±1.3 vs 3.6±1.9 in IPC alone, p>0.05). These results were also confirmed by nucleosomal DNA laddering pattern. We conclude that IPC reduces lethal injury in part by decreasing apoptosis after ischemia/reperfusion and activation of the DOR may play a crucial role in IPC or morphine-induced myocardial protection.




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