[EFFECT OF SIMULATED IN VIVO PHYSIOLOGICAL ENVIRONMENT SEVERED LIMB FOSTERING SYSTEM APPLYING REMOTE ISCHEMIC CONDITIONING PERFUSION ON PRESERVING SEVERED LIMB].

OBJECTIVE: To investigate the effect of simulated in vivo physiological environment severed limb fostering system applying remote ischemic conditioning (RIC) perfusion on preserving severed limb.

METHODS: Eighteen adult Bama mini pigs (24-30 kg in weight) were randomly divided into 3 groups (n = 6). No ischemic treatment was given in group A as normal control group; the right lower limbs were completely amputated and preserved at room temperature for 3 hours to make ischemic models in groups B and C, and then the severed limbs were put into the simulated in vivo physiological environment severed limb fostering system. Continuous blood perfusion was performed in group B, and continuous blood perfusion was performed after RIC perfusion in group C. After 8 hours of perfusion, the skeletal muscle samples were harvested for the morphology observation by transmission electron microscopy. The protein levels of B-cell lymphoma-2 (Bcl-2) and Caspase-3 were detected by Western blot. The content of cytochrome C in both mitochondria and cytoplasm was determined by ELISA.

RESULTS: Transmission electron microscopy results illustrated that the muscle fibers arranged more orderly and the mitochondria swelling was slighter in group C than group B. Western blot analysis showed that the protein levels of Bcl-2 and Caspase-3 were significantly higher in groups B and C than group A (P < 0.05); the protein level of Bcl-2 significantly increased and the protein level of Caspase-3 significantly decreased in group C when compared with those in group B (P < 0.05). ELISA detection implicated that mitochondrial cytochrome C significantly reduced and cytosolic cytochrome C significantly increased in group B when compared with those in groups A and C (P < 0.05), but no significant difference was found between group A and group C (P > 0.05).

CONCLUSION: The ischemia/reperfusion-induced injury to skeletal muscle could be considerably inhibited by RIG perfusion. The simulated in vivo physiological environment severed limb fostering system applying RIC perfusion can significantly prolong the severed limb preserving time.