摘要
目的：探讨用 UW液灌注离体断肢以延长其活性的效果。方法：选择雄性健康成年 Wistar大鼠20只,体重300～400 g,手术离断双下肢。实验分两部分。实验一：随机选择10只大鼠,再随机分为灌注组（4℃下采用 UW液自股动脉灌注离断肢体直至灌出液清亮为止）和对照组（不予灌注直接冰箱保存）,每组5只。两组均置于4℃冰箱内低温保存24 h,肢体离断后每间隔6 h取离断肢体肌肉组织行病理学观察。实验二：另10只大鼠随机分为连续灌注组（4℃下以12 ml/h的速度持续灌注 UW液24 h）和间断灌注组（4℃下于肢体离断后4、8、12、16、20和24 h以12 ml/h的速度分别灌注 UW液20 min）,每组5只。离断肢体在4℃冰箱内保存灌注24 h,收集离断后4、8、12、16、20和24 h 的灌注流出液,检测碱性磷酸酶（ALP）、丙氨酸转氨酶（ALT）、葡萄糖（GLU）含量,同时在离体12 h、24 h取肌肉组织标本,行 HE 染色观察组织结构变化,透射电镜观察其细胞超微结构的变化。结果：实验一：在各相同时间点,灌注组较对照组肌纤维水肿程度轻,断裂少,细胞较少发生横纹溶解。实验二：连续灌注组与间断灌注组比较,肌纤维均轻度水肿,细胞出现横纹溶解,线粒体肿胀,发生破裂及空泡化现象,两组病理及超微结构改变程度相似,各时间点两组之间 ALP、ALT、GLU 差异无显著性（P＞0.05）。结论：使用 UW液对离断肢体进行灌注后低温保存效果更好。UW液24 h 间断灌注与24 h 连续灌注两种方法对离断肢体的保护效果相当。
Objective：To investigate the effect of University of Wisconsin （UW）solution in prolonging the vital-ity of amputated limbs. Methods：Both hindlegs were amputated in 20 adult male and healthy Wistar rats weighing of 300-400 g. In part one of the experiment,10 Wistar rats were randomly divided into 2 groups （5 rats in each group）.The femoral artery of each of the amputated limbs was perfused with UW solution （4 ℃）until the liquid escaping from the femoral vein became clear. The amputated limbs of the control group were devoid of any special treatment. Limbs in both groups were preserved in the refrigerator at 4 ℃. Samples of tissue were taken for path-ological examination every 6 hours after the amputation. In part two of the experiment,10 Wistar rats were ran-domly divided into 2 groups （5 rats in each group）.In the continuous perfusion group the amputated limb was per-fused with UW solution continuously for 24 hours at 4 ℃ with the speed of 12 ml/h.In the intermittent perfusion group the amputated limb was perfused with UW solution periodically with 12 ml/h for 20 minutes every 4 hours for 24 hours after amputation. Limbs in both groups were preserved in the refrigerator at 4 ℃. The irrigation fluid from each perfusion was collected to determine the contents of alkaline phosphatase （ALP）,alanine aminotrans-ferase （ALT）and glucose,and muscles were sectioned to observe the changes in muscles harvested every 1 2 hours under light microscope and electron microscope after the limbs had been amputated. Results：In the experiment one,the degree of edema,fragmentation,dissolution of striped muscles were less marked in perfusion group than that of the control group at the same time point. In the experiment two,amputated limbs perfused with UW solu-tion continuously showed the same changes in structure and microstructure as the group treated with UW solution discontinuously,including fiber edema,fragmentation and horizontal dissolution in muscle.There were also swell-ing and fracture of mitochondria. There was no significant difference in contents of ALP,ALT and GLU between two groups （P〉0.05 ）. Conclusions：Perfusion with UW solution and cryopreservation has better protective effects on dismembered limbs compared with that undergoing cryogenic preservation only.The protective effects were same between continuous and periodic perfusion with UW solution on dismembered limbs for 24 hours.
出处
《感染．炎症．修复》
2014年第2期84-87,F0003,共5页
Infection Inflammation Repair
基金
国家自然科学基金资助项目(81272123)
关键词
离断肢体
器官保存
灌注
UW液
Amputated limb
Organ preservation
Perfusion
UW solution
