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Background: Drug addiction is one of the social damages that researchers have tried to find suitable strategies for prevention and treatment of it. Meanwhile, short-term physical activity is acceptable parameter for treatment and prevention of addiction. On the other hand, prefrontal cortex, for a long time as mediating, is involved in many of the complex behavioral responses, especially those associated with addiction and drug abuse. In this paper, the effects of short-term physical activity (treadmill running) on withdrawal symptom with or without mPFC area have been evaluated. Methods: This experimental study was done on Wistar rats weighing 250 ± 300 g separated to four groups: control, shame, test 1 and test 2; one of the test groups had surgery and lesion on mPFC area. For studying the effects of short-term physical activity, we used treadmill with adjustable speed of 17 m/minute and a 15-degrees slope; after the injection of three doses of morphine, 10, 20, 40 mg in 9 days and running on the treadmill, in the tenth day, the symptoms of addiction were evaluated. Findings: The morphine injection and running on the treadmill without mPFC region destruction tended to decreasing morphine tendency. There was a significant difference between the rats, with or without mPFC area, received morphine but did not run on the treadmill in morphine tendency (P < 0.05). Conclusion: It seems that short-term physical activity (treadmill running) decreases the tendency of using morphine with mPFC area in comparison with other groups, especially with group operated in their mPFC area.