Organ Transplantation from non-heart beating

无心跳器官移植

• 批准号: 10557112
• 负责人: KURODA Yoshikazu
• 金额: $ 7.04万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557112/
• 关键词: Pancreas transplantation; Warm ischemic injury; Two-layer method; UW simple cold storage; Protein synthesis; Stress protein

1. Based on our previous canine experiments, pancreas grafts subjected to 90 min warm ischemia, which cannot survive after implantation with no treatment, could be successfully transplanted after 5-hour preservation by the two-layer method. On the hypothesis that protein synthesis during the two-layer preservation would be associated with this graft resuscitation, the following experiments were undertaken. We preserved canine pancreases damaged by 90-min warm ischemia using the two-layer method (perfluorochemical/UW solution) for 5 hrs at 20℃. Intragraft DNA, RNA and protein synthesis were quantitated durins preservation by determining incorporation of tritiated thymidine, tritiated uridine and tritiated leucine, respctively, added in this preservation solution. Graft viability was judged from graft survival after autotransplantation in the cervical subcutaneous cavity. None of 7 grafts (0%) survived when they were immediately transplanted after warm ischemia, while all of 7 grafts (100%) transplanted after the two-layer preservation for 5 hrs at 20℃ survived. Significant increases in RNA and subsequent protein syntheses were observed during preservation by the two-layer method, not by UW simple cold storage. In contrast, DNA synthesis did not follow RNA and protein synthesis. We concluded that protein synthesis might be associated with the process of postischemic cellular recovery of the pancreas graft during mild hypothermic preservation by the two-layer method.2. Some reports have indicated that heat shock proteins (HSPs) have important functions in cellular recovery. We studied the role of HSPs in reducing ischemia-reperfusion injury during preservation by the two-layer method. From the results of immunohistochemical examinations and western-blotting of HSPs, HSP-60 was likely associated with functional recovery of warm ischemically damaged pancreas by the two-layer method.

1.在我们前期犬实验的基础上，对未经处理的移植胰腺，经90 min热缺血后不能存活的，采用双层法保存5 h后，移植成功。假设双层保存期间的蛋白质合成与移植物复苏相关，进行了以下实验。采用全氟化合物/UW液双层保存法，将犬胰腺在20℃下热缺血90分钟后保存5小时。在保存过程中，通过测定分别加入该保存液中的氚化胸苷、氚化尿苷和氚化亮氨酸的掺入量来定量移植物内DNA、RNA和蛋白质的合成。颈部皮下自体移植后，移植物的存活率判断移植物的活力。热缺血后即刻移植的7个移植物均无存活（0%），而20℃双层保存5 h后移植的7个移植物均存活（100%）。RNA和随后的蛋白质合成的显着增加，观察保存过程中的两层方法，而不是由UW简单的冷藏。相反，DNA合成并不跟随RNA和蛋白质合成。结论：1.在两层法亚低温保存中，移植胰腺缺血后的细胞恢复过程可能与蛋白质合成有关.热休克蛋白（Heat Shock Proteins，HSPs）在细胞恢复过程中具有重要作用。本实验采用双层膜法研究了热休克蛋白在保存过程中减轻缺血再灌注损伤的作用。免疫组化和Western-blotting结果表明，HSP-60可能与热缺血损伤胰腺的功能恢复有关。

项目成果

Fujino Y, Suzuki Y, Tsujimura T, Takahashi T, Tanioka Y, Tominaga M, Ku Y, Kuroda Y: "Possible role of heat shock protein 60 reducing ischemic-reperfusion injury in canine pancreas grafts after preservation by the two-layer method"Pancreas. in press.

Fujino Y、Suzuki Y、Tsujimura T、Takahashi T、Tanioka Y、Tominaga M、Ku Y、Kuroda Y：“热休克蛋白 60 减少两层法保存后犬胰腺移植物缺血再灌注损伤的可能作用”

Ippei Matsumoto: "Superiority of mild hypothermic (20degrees C) preservation for pancreatic microvasculature using two-layer storage method"Pancreas. 21(3). 305-309 (2000)

Ippei Matsumoto：“使用双层储存方法对胰腺微血管进行亚低温（20℃）保存的优越性”胰腺。

Matsumoto S, Sutherland DER, Hassoun AA, Kandaswamy R, Hiraoka K, Sageshima J, Shibata S, Alizona L, Tanioka Y, Kuoda Y: "Clinical application of the two-layer (University of Wisconsin solution (UW)/perfluorochemical (PFC)) method of pancreas preservation

Matsumoto S, Sutherland DER, Hassoun AA, Kandaswamy R, Hiraoka K, Sageshima J, Shibata S, Alizona L, Tanioka Y, Kuoda Y：“两层（威斯康星大学溶液（UW）/全氟化合物（PFC）的临床应用）

藤野泰宏: "二層単純浸漬保存法による膵虚血再灌流障害の軽減効果"今日の移植. 12(6). 600-603 (1999)

Yasuhiro Fujino：“两层简单浸泡保存法减少胰腺缺血再灌注损伤的效果”，《今日移植》12(6)。

谷岡康喜: "臓器保存とviability判定法"低温医学. 25(1). 30-34 (1999)

Yasuki Tanioka：“器官保存和活力测定方法”Cryogenic Medicine 25(1)。