Development of the supercooling preservation method for transplant liver graft and analysis of the mechanism of cold preservation injury

移植肝过冷保存方法的研制及冷保存损伤机制分析

• 批准号: 12671159
• 负责人: YAGI Takahito
• 金额: $ 0.58万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671159/
• 关键词: transplant organ storage; supercooling preservation; cold preservation injury; anti-freezing protein; ascorbic acid; subzero nonfreezing preservation; anti-nucleating protein; ascorbic acid 2-glucoside; liver transplantation; 肝移植; 臓器保存; 抗氷晶核蛋白

Supercooling is likely to achieve superior preserability of graft organs. In order to improve the preservation procedure of liver grafls, we attempted the subzero nonfreezing (SZNF) storage methods. In consideration of applying the SZNF storage to clinical use, we employed anti-nucleating protein (ANP), isolated from bacterium^1, as anti-freezing agent (AFA) for the prevention of concussive organ freezing.Recent studies showed that the sinusoidal endothelial cell (SEC) damages are the primary targets in cold preservation/ reperfusion injury and the apoptosis of SECs plays a critical role. For protection of SECs against supercooling preservation injury, we used ascorbic acid 2-glucoside (AA-2G) as antioxidant which is chemically stable ascorbic acid derivative.In this research, we investigated the participation of SECs apoptosis in supercold iscbemic injuiy and the efficacy of the SZNF liver storage using ANP with AA-2G in the University of Wisconsin (UW) solution.In our previous study the SZNF storage of isolated SECs failed to show hs advantages as compared with hepalocytes(HC). The problem to be solved for the SZNF liver storage is the stronger hypothermia-specific damage enhanced by supercooling. In this study the SZNF condition by itself could not achieve the better preservavility of HCs in transplanted liver due to the microcirculatoiy disturbance by SECs damage.The reactive oxygen species and free radicals appeared to play an important role in apoptosis of SECs upon cold ischemia / reperfbsion injury. The usage of AA-2G as a free radical scavenger reduced the sinusoidal damages and inhibited apoptosis in SECs enhanced by supercooling. The protective effect of AA-2G against SECs damage led to the better preservavilily of HCs as a consequence of supercooling storage in transplanted liver.The SZNF liver storage with ANP and AA-2G had a potency to surpass the conventional cold preservation

过冷可能使移植器官具有上级可移植性。为了改进肝脏移植物的保存方法，我们尝试了低温非冷冻（SZNF）保存方法。为了将SZNF保存技术应用于临床，我们采用了从细菌中分离的抗成核蛋白（ANP）^[1]作为防冻剂（AFA）来预防震荡性器官冷冻。最近的研究表明，窦状隙内皮细胞（SEC）损伤是冷保存/再灌注损伤的主要靶点，SEC的凋亡起着关键作用。为了保护SECs免受过冷保存损伤，我们使用抗坏血酸2-葡萄糖苷（AA-2G）作为抗氧化剂，这是一种化学稳定的抗坏血酸衍生物。我们在威斯康星州大学（UW）研究了SEC凋亡在超冷缺血损伤中的参与以及使用ANP和AA-2G的SZNF肝脏储存的功效。在我们以前的研究中，与肝细胞（HC）相比，分离的SEC的SZNF储存未能显示出hs优势。SZNF肝脏储存要解决的问题是过冷增强的更强的低血糖特异性损伤。SZNF本身并不能使移植肝中HC的存活率提高，这是由于SEC损伤引起的微循环障碍，活性氧和自由基在冷缺血/再灌注损伤后SEC的凋亡中起重要作用。AA-2G作为自由基清除剂的使用减轻了过冷增强的SEC的窦状隙损伤，并抑制了SEC的凋亡。AA-2G对SECs损伤的保护作用使移植肝过冷保存后的HCs保存能力增强，ANP和AA-2G联合应用的SZNF肝保存效果优于常规低温保存

项目成果

Yagi T. et al: "Problems in living donor liver transplantation in adults : postoperative management, complications, and costs."Transplantation Proceedings. 32. 2156-2157 (2000)

Yagi T. 等人：“成人活体肝移植的问题：术后管理、并发症和费用。”移植论文集。

H Matsukawa el al.: "Ascorbic acid 2-glucoside prevents sinusoidal endotbelial cell apoptosis in supercooled presented grafts in rat liver transplantation."Transplantation Proceedings. 32. 313-317 (2000)

H Matsukawa 等人：“抗坏血酸 2-葡萄糖苷可防止大鼠肝移植中过冷移植物中的肝窦内皮细胞凋亡。”移植论文集。

J Liu et al.: "Annexin V assay -proven anti-apoptotic effect of Ascorbic acid 2-glucoside after cold ischemia/reperfusion injury in rat liver transplantation"Acta Medica Okayama. (in press).

J Liu 等人：“Annexin V 测定 - 证明抗坏血酸 2-葡萄糖苷对大鼠肝移植冷缺血/再灌注损伤后的抗细胞凋亡作用”Acta Medica Okama。

Yagi T, et al.: "Successfully treated renal artery stenosis using an oral PGI2 derivative, beraprost, in cadaveric kidney transplantation."NEPHRON. 85. 188-189 (2000)

Yagi T 等人：“在尸体肾移植中使用口服 PGI2 衍生物贝前列素成功治疗了肾动脉狭窄。”NEPHRON。

H Matsukawa et al.: "Ascorbic acid 2-glucoside prevents sinusoidal endothelial cell apoptosis in supercooled preserved grafts in rat liver transplantation"Transplantation Proceedings. 32. 313-317 (2000)

H Matsukawa 等人：“抗坏血酸 2-葡萄糖苷可防止大鼠肝移植过冷保存移植物中的肝窦内皮细胞凋亡”移植论文集。