The Combined Effects Of Hyperbaric Oxygen And Mannitol To The Limb Ischemia And Reperfusion Of Different Phase

Objective These experiments were designed to investigate the occurrence and pathological changes of the limb ischemia - reperfusion injury and the combined effects of hyperbaric oxygen and mannitol to the local cytokine response , fibrinolysis response , apoptosis and ultramicro -structure and morphological changes of rat limb ischemia and re -perfusion of different phases.Methods 192 SD rats were randomly divided into various groups, underwent different phases of reperfusion after 8 hours of sustained ischemia, and treated with hyperbaric oxygen and / or mannitol. IL-1β, IL-6, IL-8, PAI-1 and tPA levels were measured in protein extracts for each experimental interval. The expression levels of cell apoptosis-related gene Bax and Bcl-2 was detected through the paraffin sections of rat gastrocnemius muscle. Electron microscope and light microscope were used to observe the ultramicrostructure and morphological changes of the skeletal muscle and capillary under the effects of different interventions. Results The Bax levels in all the intervention groups in 4h,24h,72h after reperfusion were significantly lower than the Control group. The Bcl-2 levels in MNT groups in 4h after reperfusion were significantly higher than the Control group, in HBO and MNT-HBO groups in 4h,24h after reperfusion were significantly higher. The IL-1βlevels in all the intervention groups in 4h,24h,72h after reperfusion were significantly lower than the Control group. The IL-6 levels in MNT groups in 24h,72h after reperfusion were significantly lower than the Control group,in HBO groups in24h,72h after reperfusion were significantly lower and in MNT-HBO groups in 4h,24h,72h after reperfusion were significantly lower. The IL-8 levels in MNT groups in 4h,24h,72h after reperfusion were significantly lower than the Control group,in HBO and MNT-HBO groups in 24h,72h after reperfusion were significantly lower. The PAI-1levels in all the intervention groups in 24h,72h after reperfusion were significantly lower than the Control group. The tPA levels in MNT groups in 4h,24h,72h after reperfusion were significantly higher than the Control group, in HBO and MNT-HBO groups in 24h,72h after re -perfusion were significantly higher. The combined effects of hyperbaric oxygen and mannitol that the apoptosis ,the local cytokine response and fibrinolysis response were inhibitted were significantly better than singlly-using hyperbaric oxygen or mannitol (P <0.05). The role of hyper -baric oxygen was significantly better than mannitol (P <0.05).In HBO groups and MNT groups,the cellular swelling of skeletal muscle and the local cytokine response were significantly lower, microvascular throm -bosis in capillary vessel,the collapsed sarcomere , shattered plasma membrane, swelled mitochondrial were all less than in Control groups. Also, The combined effects of hyperbaric oxygen and mannitol that the ultramicrostructure and morphological changes were inhibitted were significantly better than singlly-using hyperbaric oxygen or mannitol.Conclusions The combined effects of hyperbaric oxygen and man -nitol had a more significant role in the protection to the limb ischemia and reperfusion of different phase.