The PUFA-derived lipid mediator response shifts from pro-inflammatory to inflammation resolution over time and may be modified by regular moderate exercise. This pre-post-test study aimed to compare the expression of PTGES2 (COX2) and ALOX15 in leucocytes and the plasma 5- and 15-HETE, 18-HEPE and 17-HDHA responses after unaccustomed resistance exercise between 18–35-year-old male recreational runners (n=18) and less-active controls (n=15). One repetition maximum (1RM) was determined for squats, 45° leg presses and leg extensions. Subsequently three sets of 8–10 repetitions were performed at 80% 1RM and blood collected over 72 hours. PTGES2 and ALOX15 expression changed over time in runners (P=0.016, P=0.007) but not controls (P=0.631, P=0.539). 5- and 15-HETE changed over time in runners (P<0.001, P=0.022), but not controls (P=0.457, P=0.985). 18-HEPE changed in runners and controls (P<0.001, P=0.024), 17-HDHA changed borderline in runners (P=0.076). In conclusion, pro-inflammatory and inflammation-resolving lipid mediators may respond sooner and more robust in recreational runners than less-active controls after strenuous resistance exercise.