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Objective: To study the pretreatment effect of adenosine on NF-κB nuclear activity in ischemia/reperfusion myocardium in rats. Methods: Eighteen healthy male S-D rats were randomly divided into three groups. Group I was the control group. The other two groups were subjected to 30 minutes of ischemia, followed by 2 hours of reperfusion. In group III, adenosine was given 40 μg*kg-1*min-1 30 minutes before coronary artery occlusion. The NF-κB in nuclear was extracted and measured with western blot analysis. TNF-α levels in myocardium were measured with enzyme-linked immunosorbent assay (ELISA) system. All the data were recorded with mean±SEM, differences at the 95% confidence level were considered significant. Results: NF-κB activity in the nucleus significantly increased after ischemia/reperfusion and TNF-α levels changed. Adenosine significantly inhibited NF-κB activity in nucleus, and concomitantly decreased the level of TNF-α in myocardium. Conclusions: Adenosine modulation of NF-κB activation may be the cellular molecular mechanism of decreasing of TNF-a. The cardioprotective action of adenosine may be involved in the differential modulation of NF-κB activation during myocardial ischemia/reperfusion.