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Background and Aim: The prevalence of overweight and obesity is increasing dramatically due to unhealthy lifestyles. This study aimed to determine the effects of six weeks of total-body resistance exercise (TRX) combined with purslane supplementation on serum levels of brain-derived neurotrophic factor (BDNF), adropin, and nesfatin in overweight girls. Methods: In this semi-experimental study, 32 overweight female students aged 16 to 18 years (BMI > 25) from Ganaveh County, Bushehr Province, Iran, were randomly selected. The participants were divided into four groups: exercise, supplement, exercise + supplement, and placebo. The exercise groups performed TRX exercises consisting of 9 to 11 movements for six weeks, with three sessions per week, each lasting 60 minutes. Purslane supplementation was administered as two 500 mg capsules of purslane (Perpin Ala) taken daily after breakfast and lunch. Serum levels of BDNF, adropin, and nesfatin were measured using ELISA at the beginning and end of the six-week study. Results: The serum levels of BDNF in the exercise + supplement group showed significant differences compared to the exercise, supplement, and control groups (P < 0.05). Additionally, the serum levels of nesfatin were significantly different between the exercise + supplement group and the control group (P < 0.05). However, there were no significant differences in serum levels of adropin among the groups (P > 0.05). Conclusion: The findings suggest that a combination of TRX exercises and purslane supplementation over six weeks positively affects BDNF and nesfatin levels in young girls, potentially aiding in weight reduction. Further research is needed to explore this area more comprehensively.