<p>Prostaglandin E-major urinary metabolite (PGE-MUM) may be a useful biomarker for assessing muscle inflammation caused by physical exercise. To explore this, both male and female volunteer university students (<i>n</i> = 18) performed gradual loading treadmill tests. Peripheral blood and urine samples were obtained before (T1) and immediately after physical exercise (T2), and 2&#xa0;h later (T3) for measurement of PGE-MUM. PGE-MUM∙Cr (corrected by urinary creatinine) showed a significant stepwise increase from T1 to T3 in females (<i>p</i> = 0.0171) and the same tendency without significance was observed in males (<i>p</i> = 0.1533). Serum myoglobin showed the same trend, increasing from T1 to T3 in both males and females (<i>p</i> = 0.1690, 0.0443 respectively); Of note, females had a significant correlation between myoglobin and PGE-MUM∙Cr, (<i>p</i> = 0.0456), suggesting the involvement of muscle damage, but not between PGE-MUM∙Cr and peripheral leukocytosis after physical exercise. PGE-MUM∙Cr had no independent association with serum myoglobin after accounting for total muscle weight and maximal oxygen uptake/BW(VO₂max/BW). Observable gender differences, with males showed significantly higher values of PGE-MUM∙Cr, myoglobin and creatinine in serum, and VO₂max/BW than females, suggest a possible relation of PGE-MUM∙Cr with muscle weight and inflammation.</p>