持续低温灌注转运系统（AirdriveTM）影响犬肾保存的实验研究

摘要

目的：采用荷兰 INDES 公司生产的 AirdriveTM持续机器灌注仪,探讨其对犬肾低温缺血损伤的保护作用。方法使用健康草犬1只,全身麻醉后获取双侧肾脏,2只肾脏分别用普通低温保存和持续机器灌注低温保存。普通低温保存的肾脏浸泡在冰水混合的器官保存液中,并置于4℃的低温房间内存放。持续机器灌注保存的肾脏则置于 AirdriveTM持续机器灌注仪中,该仪器置于室温下存放。于不同时间点获取肾组织分别进行组织病理学、电镜及线粒体活性检查,比较两组的不同。结果1)组织切片：0~24 h 肾组织的结构基本相似。但48 h 后,普通低温保存的肾脏出现较明显的改变,其中肾小管上皮细胞的改变比肾小球更为明显。2)电镜：48 h 后,普通低温保存的肾脏出现较明显的形态学改变,肾小管及肾小球的基膜厚度不均一,细胞内褶明显减少且紊乱。线粒体基质的电子密度明显减低,嵴数量减少且排列紊乱。3)线粒体活性：随着低温保存时间延长,持续机器灌注组和普通低温保存组肾皮质线粒体功能均明显降低。保存12~48 h,普通低温保存组Ⅲ态呼吸呼吸耗氧速率下降明显且显著低于持续机器灌注组(P<0.05)；48 h 普通低温保存组较持续机器灌注组Ⅳ态呼吸耗氧显著增加(P<0.05),余时间点2组间差异无统计学意义；48 h 持续机器灌注组较普通低温保存组皮质呼吸控制率(respiratory control ratio,RCR)、P/ O 显著增高,余时间点2组间比较差异无统计学意义。结论随着保存时间的延长,持续低温灌注转运系统(Airdrive)对犬肾低温保存的效果明显优于普通低温保存。随着使用边缘器官的增多,持续机器灌注的应用必将得到进一步的推广以及具备保护供体器官的能力。%Objective To evaluate the protective effect of continuous hypothermic machine perfusion transport system(AirdriveTM ) on cold ischemic injury of canine kidney. Methods Two kidneys of a healthy dog were taken out after general anesthesia. One kidney was stored using common cold preservation(CCP group) by immersing in the organ preservation solution, mixed with water and ice, and kept in a cold room at 4 ℃ . The other kidney was stored using continuous machine perfusion preservation(CMP group) and was placed into the AirdriveTM continuous machine perfusion device at room temperature. The renal tissues were examined by histopathology, electron microscopy, and mitochondrial activity was checked at different time points. Results Histologic sections showed that the structures of renal tissues of the 2 kidneys were similar during first 24 hours. After 48 hours, CCP group showed more pronounced changes, as the renal tubular epithelial cells were more obvious than those in the glomeruli. After 48 hours, CCP group showed more obvious morphological changes. The tubular and glomerular basement membranes had uneven thickness. The cell infoldings were obviously reduced and disordered. The electron density of the mitochondrial matrix was significantly decreased. The cristae were reduced and disarranged. Oxygen consumption rate of state III and IV respiration in CCP group decreased after 12 ~ 48 hours and increased at 48 hours, respectively, when compared to continuous machine perfusion(CMP) group(P<0. 05). At 48 hours, cortex respiratory control ratio(RCR) and phosphorus oxygen ratio(P/ O) were significantly higher in the CMP group, compared to the CCP group. However, there was no significant difference between two kidneys at any other time points. Conclusion With prolonged storage time, the continuous hypothermic machine perfusion transport system showed better effect than that of common cold preservation on canine kidney. With increased use of marginal organs, the CMP will be further popularized clinically, and it has the ability to achieve the purpose of protecting the donor organs.