The effect of forced treadmill running on the level of some cytokines in skeletal muscles of mice with a model of type II diabetes mellitus was studied. The mouse model was developed using a 12-week high-fat diet; physical loading in the form of forced treadmill running was applied for 4 weeks. The concentration of myokines in m. gastrocnemius was determined by an enzyme-linked immunosorbent assay (ELISA). Diabetes formation in mice was accompanied by changes in the skeletal muscle concentration of two pro-inflammatory cytokines: an increase in IL-6 and a decrease in IL-15. Forced treadmill running loads had differential effects on the level of myokines in healthy and sick mice. In healthy animals, there was a decrease in IL-6 and IL-15 concentrations and an increase in the leukemia inhibitory factor (LIF) concentration in skeletal muscles after 4 weeks of regular forced running. At the same time, in diabetic mice, IL-6 and IL-15 concentrations increased, while the LIF concentration decreased after forced running loads. The NAP3 concentration in skeletal muscles was found to be insensitive to both the development of type II diabetes mellitus and regular forced treadmill running.