脉络宁对体外模拟体内生理环境寄养断肢系统缺血/再灌注损伤模型的作用

Effect of Mailuoning on an ischemia/reperfusion injury model in the system of simulated in vivo physiological environment for amputated limbs

背景:减少体外模拟体内生理环境寄养断肢系统的缺血再灌注损伤,可以延长断肢的保存时间,提高断肢的再植成功率。目的:观察脉络宁对体外模拟体内生理环境寄养断肢系统缺血再灌注损伤的影响。方法:健康成年雄性巴马小型猪18只,随机分成冷藏组、血液灌注组和脉络宁联合血液灌注组,每组6只。将每只猪的左后肢离断室温保存3 h后,冷藏组置于4℃冷藏,血液灌注组用血液灌注、脉络宁联合血液灌注组用脉络宁联合血液灌注。灌注6 h后,采用透射电镜分别观察各组离断肢体骨骼肌形态学变化;实时定量RT-PCR检测离断肢体骨骼肌Caspase3和白细胞介素1βmRNA表达。结果与结论:透射电镜结果显示,血液灌注组、脉络宁联合血液灌注组均较冷藏组肌纤维整齐、肌节完整、线粒体肿胀程度轻,且脉络宁联合血液灌注组情况优于血液灌注组;实时定量RT-PCR检测结果显示,3组的caspase3和白细胞介素1β的表达均高于正常,同时血液灌注组和脉络宁联合血液灌注组的caspase3和白细胞介素1β的表达水平明显低于冷藏组,且脉络宁联合血液灌注组表达水平低于血液灌注组。结果提示在体外模拟体内生理环境寄养断肢系统中,脉络宁可显著减轻断肢骨骼肌细胞的缺血再灌注损伤,延长断肢的保存时间。

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.