体外模拟体内生理环境寄养断肢系统采用远端缺血处理灌注方法对保存断肢的作用研究  

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

作　　者：李腾飞[1] 王江宁[2] 高磊[2] 尹叶锋[2] 

机构地区：[1]首都医科大学附属北京朝阳医院,北京100020 [2]首都医科大学附属北京世纪坛医院修复重建外科

出　　处：《中国修复重建外科杂志》2016年第1期91-94,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家自然科学基金资助项目(81473502)~~

摘　　要：目的研究体外模拟体内生理环境寄养断肢系统采用远端缺血处理（remote ischemic conditioning,RIC）灌注方法对保存断肢的作用。方法选取健康成年雄性巴马小型猪18只,体质量24~30 kg,随机分为3组（n=6）。A组为正常对照组,未经任何缺血处理;B、C组采用右后肢完全离断并置于室温保存3 h建立骨骼肌缺血模型,之后将断肢接入体外模拟体内生理环境寄养断肢系统,B组进行持续血液灌注,C组行RIC灌注后再持续血液灌注。灌注8 h后取各组骨骼肌标本,行透射电镜形态学观察;采用Western blot检测组织中B淋巴细胞瘤2（B-cell lymphoma-2,Bcl-2）及半胱氨酸蛋白酶3（Caspase-3）蛋白水平,ELISA检测线粒体及细胞基质内细胞色素C含量。结果透射电镜观察示,与B组相比,C组骨骼肌纤维排列整齐紧凑,线粒体肿胀程度轻;Western blot检测示,B、C组Bcl-2及Caspase-3蛋白水平均较A组明显升高;与B组比较,C组Bcl-2蛋白水平显著升高,而Caspase-3蛋白水平显著降低;组间比较差异均有统计学意义（P〈0.05）。ELISA检测示,与A、C组比较,B组线粒体内细胞色素C含量显著降低,但细胞基质内含量明显升高,比较差异均有统计学意义（P〈0.05）;A、C组间比较差异均无统计学意义（P〉0.05）。结论 RIC灌注方法可以显著抑制缺血再灌注引起的骨骼肌损伤,体外模拟体内生理环境寄养断肢系统采用RIC灌注可能起到延长断肢体外寄养时间的作用。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 RIC perfusion. The simulated in vivo physiological environment severed li

关 键 词：模拟体内生理环境寄养断肢系统 远端缺血处理 缺血再灌注损伤 猪 

分 类 号：R658.1[医药卫生—外科学]