This study investigated the time course of changes in heat shock protein (HSP) 72, muscle weight and cross-sectional area of muscle fibers after injection of bupivacaine (BPVC) into the soleus muscle. Adult male Wistar rats (n=36) were anesthetized by pentobarvital sodium and 0.5 ml of BPVC was injected into the left soleus muscle, whereas the contralateral right soleus muscle was served as the control. Bilateral soleus muscles were dissected at 1, 2, or 4 days or 1, 2, or 4 weeks after the injection. Muscle fibers were disrupted at days 1-2 after BPVC injection and regenerated fibers that have centrally located nuclei in cytoplasm appeared 4 days after the injection. The cross-sectional area of regenerated fibers gradually increased during 4 days-4 weeks after the injection. The soleus muscle weight decreased until 1 week after the injection, then gradually increased during 1-4 weeks of recovery. The content of HSP 72 was drastically decreased in the soleus muscle 1 day after BPVC injection (p<0.01), then gradually increased during 2 days-4 weeks of recovery, and returned to the control level at 4 weeks of recovery. A similar time course of change was observed for the cross-sectional area of the regenerating fibers and HSP 72 expression. These results suggest that the BPVC injection induces disruption of muscle fibers and degradation of HSP 72, however, regeneration of muscle fibers indicated by increases in cross-sectional area occurs with increases in HSP 72 content during 4 weeks of recovery period.