肢体缺血再灌注损伤的新型动物模型制作

目的]用充气止血带制作肢体缺血再灌注损伤的新型动物模型,研究其对周围神经和骨骼肌损伤的影响.方法]选择健康新西兰大白兔6个月龄,30只,体重(3.5 ±0.3) kg,雌雄不限,在家兔左侧后肢环扎充气止血带,于不同时间点松开,造成左侧后肢缺血再灌注损伤的模型.随机分为3组,每组10只.A组:对照组,B组:缺血2h,C组:缺血4h.对照组不扎充气止血带,第1、2、3、4、5、6h检测肢体的神经电生理学指标,B组、C组于再灌注(松开止血带,血供恢复后)的1、2、3、4、5、6h检测肢体的神经电生理学指标,A组于第6h观察骨骼肌的形态,B、C组于再灌注(松开止血带,血供恢复后)的第6h观察骨骼肌的形态,每组于术后第5d评估左侧后肢的行走功能.结果]随着缺血后再灌注时间的延长,B、C和A组相比较,周围神经的潜伏期延长、波幅降低,传导速度降低,三组之间的潜伏期、波幅、传导速度差异均有统计学意义(P＜0.05),光镜观察骨骼肌可见(B、C组):横纹紊乱、肌纤维断裂、间质血管扩张充血、大量中性粒细胞浸润.结论]经过缺血期和再灌注损伤的交互作用后,肢体的功能性损伤进一步加重,出现了不可逆的病损.该模型制作对动物的损伤较小,更贴近临床.

New animal model making of limb ischemia reperfusion

Objective]Pneumatic tourniquet was used to make limb ischemia reperfusion of animal model to investigate the injury of peripheral nerve and the skeletal muscle.Method]Thirty health 6-month-old New Zealand white rabbits(weighing 3.5±0.3 kg,male or female) were choosen,the left hind limb in rabbits was circled with inflatable tourniquet release at different times,resulting in the left hind limb ischemia-reperfusion injury model.Animals were randomly divided into 3 groups of 10 each,group A: a control group B: ischemia for 2 hours,group C: ischemia for 4 hours.Except group A all the groups were circled with inflatable tourniquet.Each group were measured with indicators of limb electrophysiology reperfusion after 1 hour,2 hours,3 hours,4 hours,5 hours,6 hours.The muscle of group A was observed by morphology after 6 hours.The muscle of group B and C were observed by morphology 6 hours after reperfusion.The walking function of left hind limb of every rabbits was evaluated 5 days after surgery.Result] With the longing of ischemia-reperfusion time,the incubation period of the peripheral nerves extended,amplitude lowered and conduction velocity decreased in groups B,C compered with group A.The incubation,amplitude and conduction velocity among three groups had statistically significant difference(P<0.05).Skeletal muscle under microscopy(group B and C) showed stripes disorders,muscle fiber rupture,interstitial vascular dilatation and congestion,and a large number of neutrophils infiltration.Conclusion] After the interaction of ischemia and reperfusion,the physical damage cound further increase the functionality,or even lead to irreversible lesions.The animal model making has less damage to animals,and is similar to clinical practice.