BACKGROUND:Reducing the ischemia-reperfusion injury in the system of simulated in vivo physiological environment for amputated limbs, can extend the preservation time and improve the replantation success rate of amputated limb. OBJECTIVE:To investigate the effect of Mailuoning on the ischemia-reperfusion injury in the system of simulated in vivo physiological environment for amputated limbs. METHODS:The left hind legs harvested from 18 healthy adult male Bama mini-pigs were randomly divided into three groups:cold storage group, blood perfusion group, Mailuoning combined blood perfusion group, with six pigs in each group. After the left hind legs of each pig in al groups were amputated and stored at room temperature for 3 hours, the amputated legs were placed at 4 ℃, perfused with blood, or perfused with Mailuoning combined blood. After 6 hours of perfusion, the morphology of amputated limbs was observed under transmission electron microscope. The mRNA levels of apoptosis proteins Caspase3 and inflammatory factor interleukin-1βwere detected by real time fluorescent quantitative RT-PCR.RESULTS AND CONCLUSION:Transmission electron microscopy results showed that, the muscle fibers in blood perfusion group and Mailuoning combined blood perfusion group arranged more orderly and were more complete than cold storage group, the swel ing of mitochondria was lighter. In addition, the condition in Mailuoning combined blood perfusion group was better than in blood perfusion group. RT-PCR results showed that the expression of Caspase3 and interleukin-1βwas increased in al groups;at the same time, the expression level in blood perfusion group and Mailuoning combined blood perfusion group was significantly lower than those in cold storage group. Mailuoning combined blood perfusion group had s lower expression than blood perfusion group. In the system of simulated in vivo physiological environment for amputated limbs, Mailuoning can significantly reduce the ischemia/reperfusion injury of skeletal muscle cells and prolong the preservation time of amputated limbs.