Background and aim: The aim of this study was to investigate the effect of aerobic exercise and plant hydro-alcoholic extract on serotonin levels and headache indices in patients with chronic tension-type headache (CTTH). Materials and methods: In this practical study, 18 adult female patients CTTH were randomly divided into two equal groups: exercise and herbal hydro-alcoholic extract. Aerobic training was performed three sessions a week, each session lasting 75 minutes for eight hours. Herbal extract was taken in capsule form. The dose of each capsule was 280 mg, which was taken three times a day. Serotonin levels, headache indices (severity, duration, and frequency of headache), anxiety, depression, use of synthetic drugs, and aerobic capacity were evaluated at baseline and at the end of the study. Paired-Sample and independent t-test was used to compare intra-group and intergroup changes and a significance level of 0.5 was used. Results: The intra-group results showed a significant increase in serotonin, aerobic capacity (exercise group only), and a significant decrease in anxiety, depression, intensity, frequency and duration of headaches (only in the exercise group) and the use of synthetic drugs. In the comparison of two groups, there was a significant difference between headache frequency, use of synthetic drugs, anxiety and aerobic capacity. Conclusion: Aerobic exercise and hydro-alcoholic extract of valerian and lemon balm plants can be a suitable treatment supplement for women suffering from CTTH but the effect of two plants was better in improving headache.