Sex and hormonal influences on stress responses in 55 individuals with burnout symptoms were examined using psychological assessments, cortisol sampling, and fMRI. Men showed stronger cortisol reactivity than women, with subgroup analyses highlighting additional effects of hormonal status in women. Neuroimaging revealed stress-related limbic and task-positive activation patterns, with sex-specific associations between cortisol responses and hippocampal engagement, while post-stress functional connectivity of the amygdala and hippocampus increased broadly but was not moderated by sex or cortisol.