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Objective(s):This study examined whether conjugated linoleic acid (CLA) supplementation affects insulin sensitivity and peroxisome proliferator-activated receptor gamma (PPAR-γ) and glucose transporter type 4 (GLUT-4) protein expressions in the skeletal muscles of rats during endurance exercise. Materials and Methods:Sprague-Dawley male rats were randomly divided into HS (high-fat diet (HFD) sedentary group, n = 8), CS (1.0% CLA supplemented HFD sedentary group, n = 8), and CE (1.0% CLA supplemented HFD exercise group, n = 8). The rats in the CE swam for 60 min a day, 5 days a week for 8 weeks. Results:The serum glucose and insulin contents and homeostasis model assessment of insulin resistance (HOMA-IR) value of the CS and CE were significantly decreased compared to those of the HS. The PPAR-γ protein expressions in the soleus muscle (SOM) and extensor digitorum longus muscle (EDL) were significantly higher in the CE than in the HS. In addition, the PPAR-γ protein expression in the SOM of the CS was significantly higher than that in the HS. On the other hand, the GLUT-4 protein expression of the SOM in the CE was significantly higher compared to that in the HS. However, there was no significant difference in GLUT-4 protein expression in the EDL among the groups. Conclusion:CLA supplementation with/without endurance exercise has role in improvement of insulin sensitivity. Moreover, when CLA supplementation was accompanied by endurance exercise, the PPAR-γ protein expression in SOM and EDL and the GLUT-4 protein expression in SOM were enhanced compared with the control group.