Delayed-onset muscle soreness (DOMS) is a common but displeasing event induced by excessive muscle use or unaccustomed exercise and characterized by tenderness and movement-related pain in the exercised muscle. Thermal therapies, either icing or heating applied to muscles immediately after exercise, have been used as therapeutic interventions for DOMS. However, the mechanisms of their analgesic effects, and physiological and metabolic changes in the muscle during thermal therapy, remain unclear. In the present study, we investigated the effects of both thermal treatments on mechanical hyperalgesia of DOMS and physiological and muscle metabolite changes using the rat DOMS model induced by lengthening contraction (LC) to the gastrocnemius muscle. Heating treatment just after LC induced analgesic effects, while rats with icing treatment showed mechanical hyperalgesia similar to that of the LC group. Furthermore, increased physiological responses (e.g., muscle temperature and blood flow) following the LC were significantly kept high only in the rats with heating treatment. In addition, heating treatment increased metabolites involved in the improvement of blood flow and oxidative metabolisms in the exercised muscle. The results indicated that heating treatment just after LC has analgesic effects on DOMS, which might be mediated partly through the improvement of muscle oxidative metabolisms by changes in metabolites and elevated physiological responses.NEW & NOTEWORTHY Physiological effects of thermal therapy in the muscle and its mechanisms of analgesic effects remain unclear. The results indicated that heating, but not icing, treatment just after lengthening contractions induced analgesic effects in the rat muscle. Increases in hemodynamics, muscle temperature, and metabolites such as nicotinamide were more prominent in heating treatment, consistent with improvement of muscle oxidative metabolisms, which might reduce chemical factors to induce mechanical hyperalgesia.