The purpose of this study is to contribute to efforts to understand the pathophysiology of migraine and create an alternative perspective for prophylaxis and treatment of migraine attacks. Acupuncture treatment was applied to the 22 volunteer migraine patients. The acupuncture treatment consists of 5 sessions with 2 sessions per week. Blood samples have been collected before performing acupuncture, after the 1st session and after the 5th session of the acupuncture. And for the control group blood samples were collected only once. In our study the mean serum NO levels in healthy people were (3.58 ± 0.53) while patients with migraine group were (5.55 ± 0.70) respectively. Serum NO levels were 55% higher in migraine group compared to the control group. The NO concentration also decreased after 5th session (30%) (p < 0.05). After the 1st sessions NO level decreased 4.86% in the migraine group. After the 5th session NO level decreased 30.63% in the migraine group. After the 5th session NO level decreased 27.08% compared to 1st session in the migraine group. After the 1st session of acupuncture treatment, level of NO in migraine group were 4.86% and it is reduced but statistically not significant. After the 5th acupuncture treatment serum NO levels were reduced to (3.85 ± 0.62) 30.63% in migraine group and were statistically lower (p < 0.05). After the 5th session NO level decreased 27.08% compared to 1st session in the migraine group. In this study, acupuncture treatment is appeared to be effective by lowering the levels of serum NO and acupuncture has cumulative effects. Although there are earlier studies showing the effect of NO in migraine, this has been the 1st study in this field, which shows the effect of acupuncture on NO in migraine patients.