Abstract Background University students are faced with several stress factors affecting their mental health. Therefore, the first year at university is a period that calls for careful attention and research. Aim The aim of this study is to evaluate the effect of laughter yoga on mental symptoms and cortisol levels in nursing students. Methods This study is a randomized controlled study employing a pre‐/post‐test design with a control group. A total of 75 healthy university students were assigned to the intervention group and control group. The Brief Symptom Inventory was applied to both groups before Session 1 and after Session 8. Saliva samples were taken from the students to measure their cortisol levels before and after each session. Results Evaluation of the mean scores obtained from the Brief Symptom Inventory before and after the intervention showed a significant decrease in the scores between groups ( P < 0.05). In three out of the eight sessions, there was a significant decrease in the intervention group compared with the control group regarding the mean values of pre‐test and post‐test salivary cortisol levels ( P < 0.05). Conclusion Laughter yoga can provide an effective means to help first‐year nursing students cope with stress and reduce mental symptoms.