Objective We evaluated cardiovascular autonomic reflexes and serum fibroblast growth factor 21 (FGF21), a surrogate marker of mitochondrial function, in a cohort of overweight and normal-weight adults (n = 42). Methods Indices of autonomic function were monitored during supine rest, autonomic reflex tests and submaximal clinical exercise test, with heart rate variables and blood pressure measured with an automatic system. Results Markers of sympathetic dominance were accentuated in overweight adults, including elevated resting low-frequency to the high-frequency ratio for heart rate variability (203 ± 227 vs. 96 ± 42; p = .01), and handgrip diastolic blood pressure (36 ± 15 mmHg vs. 25 ± 12 mmHg; p = .01). A weak non-significant trend has been found for a negative correlation between blood pressure responses to isometric handgrip test and FGF21 in the overweight group (r = −0.37; p = .09). Conclusions Excess body weight appears to trigger sympathetic overactivity in overweight adults, yet autonomic dysregulation might not be associated with notable changes in serum FGF21.