常温机械灌注改善猪心脏死亡供体捐献供肝质量和胆汁分泌功能的研究

Nomorthermic machine perfusion improves the quality and bile secretion of donation after cardiac death liver

目的 探讨体外常温机械灌注对心脏死亡供体捐献（DCD）离体供肝活性保护和质量改善的作用.方法 15头小型猪随机均分为活体组、热缺血组和冷保存组.活体组直接获取供肝,冷保存组与热缺血组猪诱导心脏死亡90 min后获取.获取后,冷保存组供肝4℃保存4h,热缺血组与活体组使用常温机械灌注4h,各组供肝均全血机械复流1h.随后留取复流液及新生胆汁检测肝功能,切取肝及胆总管组织行苏木素-伊红（HE）染色.取活体组及热缺血组各2例供肝行18F标记脱氧葡萄糖（18F-FDG）正电子发射计算机断层显像（PET-CT）扫描,比较灌注前后平均标准摄取值（SUVmean）值.结果 热缺血组与冷保存组间的谷丙转氨酶（ALT）、乳酸脱氢酶（LDH）、γ-谷氨酰转肽酶（GGT）结果差异无统计学意义（P＞0.05）.热缺血组新生胆汁[（2.67±1.05） ml]多于冷保存组[（0.87±0.34） ml,P＜0.05].热缺血组新生胆汁中总胆红素浓度[（466.15±156.11） μmol/L]高于冷保存组[（160.79±36.69） μmol/L,P＜0.05].两组肝细胞均出现广泛水肿及肝细胞变性,胆道上皮完整性严重破坏,但冷保存组损伤更为严重.正电子发射计算机断层显像（PET-CT）结果显示热缺血组机械灌注后SUVmean值较前升高.结论 体外常温机械灌注可明显改善DCD供肝糖代谢和胆汁分泌功能,但对于缺血再灌注损伤无明显减轻作用.

Objective To investigate the protection and quality improvement of the ex vivo no-morthermic machine reperfusion for donation after cardiac death （DCD） liver.Methods Fifteen pigs were randomly divided into 3 groups：living donor group,warm isehemia group,and cold preservation group.There was no ischemia injury in living donor group,and in cold preservation group and warm ischemia group cardiac death was induced 90 min before procurement.Livers from cold preservation group were stored at 4℃ for 4 h,and nomorthermic machine perfusion was used in warm ischemia group and living donor group for 4 h with perfusate.Each group was machine reperfused for 1 h with autologous blood before blood samples were collected.Liver tissue and common bile duct samples were fixed for hematoxylin and eosin （HE） staining.F-18-fluoro-deoxy-D-glucose positron emission tomography （18 F-FDG PET） scan was performed on randomly selected 2 cases from both living donor group and warm ischemia group to compare the SUVmean before and after perfusion.Results Significant difference was found in bile output [（2.67 ± 1.05） vs.（0.87 ± 0.34） ml] and total bilirubin concentration [（466.15 ± 156.11） vs.（160.79 ±36.69） μmol/L] between the two groups （P 〈0.05）.No significant difference was found in the alanine aminotransferase （ALT）,lactate dehydrogenase （LDH） and GGT between two groups （P 〉 0.05）.Cold preservation group showed more severe sinusoidal space swelling,sinusoidal congestion and damage of the biliary epitheliums of common bile duct than warm ischemia group.The increased SUVmean was found after nomorthermic machine reperfusion.Conclusion According to our study,the normothermic machine perfusion improves the DCD liver function of synthesis and secretion of bile; however,it shows no significant protection against ischemia-reperfusion injury.