This study was undertaken to examine the alteration of masseter and plasma interleukin-6 after inducing occlusal interference and chronic stress. Male Wistar rats were submitted to chronic stress procedure, exposed to occlusal interference, or exposed to both mentioned procedures. Whole blood and masseter tissue were collected to determine interleukin-6 level, measured by means of ELISA. Masseter pain was evaluated using the orofacial formalin test. Masseter interleukin-6 level was significantly higher in animals submitted to combination of occlusal interference and chronic stress than in the control group (p<0.05). There was positive and significant correlation between pain response and masseter interleukin-6 level (r=0.5741; p<0.0003). No significant differences in plasma interleukin-6 level were found between groups (p>0.05), as well as no correlation with pain (p>0.05). Combination of occlusal interference and chronic stress leads to strong local reaction characterized by high levels of masseter interleukine-6. High concentrations of muscle interleukin-6 and its correlation with pain point to inflammatory background of masticatory muscle pain.