热缺血损伤猪肝脏在门静脉恒压和恒流模式下常温机械灌注的对比研究

肝脏 ›› 2020, Vol. 25 ›› Issue (8): 853-856.

热缺血损伤猪肝脏在门静脉恒压和恒流模式下常温机械灌注的对比研究

何锡然, 谭晓宇, 张琳, 邝伟键, 刘华敏, 陈素平, 郭家钘, 梁铭炬, 周伟津, 霍枫

528311 佛山广东顺德工业设计研究院(广东顺德创新设计研究院)生物医学部(何锡然,邝伟键,刘华敏,梁铭炬);中国人民解放军南部战区总医院肝胆外科(谭晓宇,霍枫);广东丁沃生医疗器械有限公司研发部(张琳,陈素平,郭家钘,周伟津)

收稿日期:2020-03-26 出版日期:2020-08-31 发布日期:2020-09-04
通讯作者: 霍枫,Email:gzhuofeng@163.com
基金资助:
军队实验动物专项科研项目[SYDW(2017)12];国家青年科学基金项目(81800556);佛山顺德区重大科技计划项目“离体肝脏机械灌注系统研发项目”(顺府办函〔2019〕42号)。

Comparative study of preservative effect of normothermic machine perfusion on porcine liver with warm ischemia injury between portal vein pressure- and flow-controlled mode

HE Xi-ran¹, TAN Xiao-yu², ZHANG Lin³, KUANG Wei-jian¹, LIU Hua-min¹, CHEN Su-ping³, GUO Jia-xing³, LIANG Ming-ju¹, ZHOU Wei-jin³, HUO Feng²

1. Department of Biomedical Sciences, Guangdong Shunde Industry Design Institute (Guangdong Shunde Innovative Design Institute), Foshan 528311;
2. Department of Hepatobiliary Surgery, General Hospital of Southern Theater Command of PLA, Guangzhou 510010;
3. Department of Research and Development, Guangdong Devocean Medical Instrument Co., Ltd, Foshan 528311

Received:2020-03-26 Online:2020-08-31 Published:2020-09-04
Contact: Huo Feng, Email: gzhuofeng@163.com

摘要/Abstract

摘要： 目的利用离体肝脏常温灌注系统初步探索门静脉恒压控制模式与门静脉恒流控制模式对热缺血损伤肝脏的保存效果差异。方法使用雌性巴马小型猪获取血液和供肝,所有供肝的热缺血时间为60 min,一组采用门静脉恒压控制模式(n=6),一组采用门静脉恒流控制模式(n=6),利用常温机械灌注设备灌注6 h,观察肝脏的灌注参数、血气生化指标以及组织病理学变化。结果灌注前,两组的冷缺血时间差异无统计学意义(P>0.05)。猪肝脏经过6 h的常温机械灌注,恒压组的肝动脉流量为(0.13±0.08) mL/min/g(肝重)比恒流组的(0.25±0.09) mL/min/g(肝重)更低(P<0.05)。两组比较,灌注液的pH、乳酸水平、葡萄糖水平差异无统计学意义(P>0.05);尿素氮水平除了在第2小时,两组比较差异无统计学意义(P>0.05)。对于肝细胞酶学和总胆红素水平,两组在灌注结束时亦差异无统计学意义(P>0.05)。恒流组的胆汁生成量为(54.47±15.63) mL,较恒压组的(6.98±3.20) mL更多(P<0.05)。两组在病理学评分的比较差异无统计学意义(P>0.05)。结论门静脉恒流模式更有利于肝动脉的供血,从而促进肝细胞进行胆汁合成,提示门静脉流量控制模式可能是更合适的灌注方式。但是这仍需要通过动物肝移植实验以及临床肝移植的数据进一步验证。

关键词: 常温机械灌注, 小型猪, 肝脏, 热缺血, 恒压控制, 恒流控制

Abstract: Objective To compare the protective effect of portal vein pressure-controlled and flow-controlled mode in porcine livers with warm ischemia by normothermic machine perfusion (NMP) system ex vivo. Methods Female Bama miniature swine were used as both blood and liver donors. The livers with 60-minute warm ischemia time were divided into 2 groups, one group was perfused in a portal vein constant pressure-controlled mode (n = 6), the other was perfused in a portal vein constant flow-controlled mode (n = 6), each group was perfused for 6 hours with NMP equipment. The perfusion parameters, blood gas parameters, biochemical indicators, and histopathology of livers were analyzed. Results There was no statistical difference in cold ischemic time between the 2 groups before perfusion (P > 0.05). After 6 hours of NMP, the hepatic artery flow in the constant pressure-controlled group [(0.13 ± 0.08) mL/min/g (liver weight)] was lower than that in the constant flow-controlled group [(0.25 ± 0.09) mL/min/g (liver weight)] (P<0.05). Between the 2 groups, there was no difference in perfusate pH, lactate level, glucose level, and blood urea nitrogen level (except for the 2nd hour) (P>0.05). There was no statistical difference in hepatocellular enzymes or total bilirubin level between the 2 groups at the end of perfusion either (P>0.05). The volume of bile produced in the constant flow-controlled group [(54.47 ± 15.63) mL] was more than that in the constant pressure-controlled group [(6.98 ± 3.20) mL] (P<0.05). In hepatic histopathological score, there were no differences between the 2 groups (P>0.05). Conclusion The constant flow-controlled mode was more beneficial to the blood supply of hepatic artery, thus promoted the bile synthesis in liver. It is suggested that the portal vein flow-controlled mode may be a better perfusion method, which requires further animal liver transplantation experiments and clinical liver transplantation trials.

Key words: Normothermic machine perfusion, Miniature swine, Liver, Warm ischemia, Constant pressure-controlled, Constant flow-controlled

何锡然, 谭晓宇, 张琳, 邝伟键, 刘华敏, 陈素平, 郭家钘, 梁铭炬, 周伟津, 霍枫. 热缺血损伤猪肝脏在门静脉恒压和恒流模式下常温机械灌注的对比研究[J]. 肝脏, 2020, 25(8): 853-856.

HE Xi-ran, TAN Xiao-yu, ZHANG Lin, KUANG Wei-jian, LIU Hua-min, CHEN Su-ping, GUO Jia-xing, LIANG Ming-ju, ZHOU Wei-jin, HUO Feng. Comparative study of preservative effect of normothermic machine perfusion on porcine liver with warm ischemia injury between portal vein pressure- and flow-controlled mode[J]. Chinese Hepatolgy, 2020, 25(8): 853-856.

参考文献

[1] 王兴强,刘懿禾,于立新, 等.肝移植治疗慢加急性肝衰竭3级临床疗效初步分析.中华肝胆外科杂志,2019,25:325-328.
[2] Jiang W, Li J, Guo Q, et al. Liver transplantation for hepatocellular carcinoma. Drug Discov Ther, 2015, 9: 331-334.
[3] Wang H, Jiang W, Zhou Z, et al. Liver transplantation in mainland China: the overview of CLTR 2011 annual scientific report. Hepatobil Surg Nutr, 2013, 2: 188-197.
[4] Laing RW, Mergental H, Mirza DF, et al. Normothermic ex-situ liver preservation: the new gold standard. Curr Opin Organ Transplant, 2017, 22: 274-280.
[5] Bral M, Galalopez B, Bigam DL, et al. Preliminary single-center canadian experience of human normothermic Ex vivo liver perfusion: results of a clinical trial. Am J Transplant, 2017, 17: 1071-1080.
[6] Nasralla D, Coussios C, Mergental H, et al. A randomized trial of normothermic preservation in liver transplantation. Nature, 2018, 557: 50-56.
[7] Ravikumar R, Jassem W, Mergental H, et al. Liver transplantation after ex vivo normothermic machine preservation: a phase 1 (First-in-Man) clinical trial. Am J Transplant, 2016, 16: 1779-1787.
[8] Mergental H, Perera M T, Laing R W, et al. Transplantation of declined liver allografts following normothermic ex-situ evaluation. Am J Transplant, 2016, 16: 3235-3245.
[9] Karangwa S A, Dutkowski P, Fontes P, et al. Machine perfusion of donor livers for transplantation: a proposal for standardized nomenclature and reporting guidelines. Am J Transplant, 2016, 16: 2932-2942.
[10] Marecki H, Bozorgzadeh A, Porte R J, et al. Liver ex situ machine perfusion preservation: a review of the methodology and results of large animal studies and clinical trials. Liver Transpl, 2017, 23: 679-695.
[11] Suzuki S, Toledopereyra LH, Rodriguez F, et al. Neutrophil infiltration as an important factor in liver ischemia and reperfusion injury. Modulating effects of FK506 and cyclosporine. Transplantation, 1993, 55: 1265-1271.
[12] Liu Q, Nassar A, Farias K, et al. Sanguineous normothermic machine perfusion improves hemodynamics and biliary epithelial regeneration in donation after cardiac death porcine livers. Liver Transpl, 2014, 20: 987-999.
[13] Liu Q, Vekemans K, Iania L, et al. Assessing warm ischemic injury of pig livers at hypothermic machine perfusion. J Surg Res, 2014, 186:379-389.
[14] Bian S, Zhu Z, Sun L, et al. Normothermic machine perfusion versus cold storage of liver in pig model: a meta-analysis. Ann Transplant, 2018: 197-206.
[15] Watson C J, Kosmoliaptsis V, Randle L V, et al. Preimplant normothermic liver perfusion of a suboptimal liver donated after circulatory death. Am J Transplant, 2016, 16: 353-357.
[16] Eshmuminov D, Leoni F, Schneider M A, et al. Perfusion settings and additives in liver normothermic machine perfusion with red blood cells as oxygen carrier. A systematic review of human and porcine perfusion protocols. Transpl Int, 2018, 31: 956-969.
[17] 李国强.中国活体肝移植小肝综合征临床诊治指南(2016版).中华移植杂志(电子版),2017,11:70-74.
[18] Vekemans K, Liu Q, Brassil J, et al. Influence of flow and addition of oxygen during porcine liver hypothermic machine perfusion. Transplant Proc, 2007, 39:2647-2651.
[19] Hart NA, Van Plaats AD, Leuvenink HG, et al. Determination of an adequate perfusion pressure for continuous dual vessel hypothermic machine perfusion of the rat liver. Transpl Int, 2007, 20: 343-352.