This study is designed to investigate the effect of ischemia post-conditioning on IR-induced skeletal muscle injury in limbs of experimental rabbits. Rabbits are randomized to one of the following three groups: sham control, ischemic reperfusion, ischemic postconditioning. The lipid peroxidation level, antioxidant enzymes activities, skeletal muscle mammalian target of rapamycin (mTOR), Bax, Bcl-2 proteins expression and Bcl-2/Bax, and HIF-1α/β-actin mRNA, interleukin-6 (IL-6)/β-actin mRNA and caveolin-3/β-actin mRNA expression were tested in the current study. The results suggested that ischemic postconditioning might decrease lipid peroxidation level, lactic dehydrogenase (LDH), creatine kinase (CK) activities, Bcl-2 proteins expression and Bcl-2/Bax, HIF-1α/β-actin mRNA expression and increase skeletal muscle antioxidant enzymes activities, Bax protein expression and IL-6/β-actin mRNA and caveolin-3/β-actin mRNA expression. These indicated that protective effect of ischemic postconditioning against IR-induced skeletal muscle injury involve into a complex molecular mechanism. Our research results may offer a theoretical guidance for therapy of related clinical diseases.