Amelioration of liver graft viability by improvement of sinusoidal microcirculation

通过改善肝窦微循环来改善肝移植物的活力

• 批准号: 18591404
• 负责人: MIYAZAWA Hideaki
• 金额: $ 2.47万
• 依托单位: Akita University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591404/
• 关键词: sinusoidal microcirculation; sinusoidal collapse; non-heart-beating donor; continuous perfusion; liver transplantation

[Introduction] Non-heart-beating donor (NHBD) livers are anticipated as an additional source of grafts to solve chronic shortage of donors. Uncontrolled NHBD graft usually suffers from an extensive injury of warm ischemia, and the transplantation therefore results in poor outcomes. To make a point for preserving NHBD livers clear, we compared post preservation damage of NHBD livers using three solutions available at present, UW, HTK, and ET-Kyoto (ETK) solutions.[Methods] Wistar rats underwent phrenotomy and livers were retrieved 60 or 30 minutes after cardiac arrest. Urokinase solution was injected via the portal vein in order to dissolve thrombi in sinusoid, then the livers were left 10 minute at room temperature. The livers were flushed by 80ml of cold UW or HTK or ETK solution, and preserved in the respective solution for 8 hours at 4℃. HBD liver without preservation was used as control. We assessed graft viability using isolated liver perfusion model. After 20-minute rewarming, li … More vers were perfused by oxygenated Krebs Henseleit Buffer. Perfusion was carried out for 60 minutes. Portal venous pressure and bile production were monitored during perfusion. AST level in the perfusate was measured. Apoptosis was evaluated by TUNEL staining.[Results] The injection of urokinase solution improved graft viability. Portal venous pressure was lower in ETK group than UW and HTK groups. ETK group produced more bile during 60-minute reperfusion than UW and HTK groups. There was significant difference between ETK group and UW group. However, bile production in ETK group was only one-third of HBD livers. The level of AST in the perfusate was lower in ETK group than UW and HTK groups. There was significant difference only in 5 minute after reperfusion between the groups of UW and ETK. In H-E staning, hepatocyte shrank markedly in the UW group, resulting in remarkable extension of sinusoidal space. On the other hand, in the HTK group, the liver architecture was better preserved. A higher osmolarity of UW solution might, in converse, facilitate the damage in the pre-injured hepatocytes in NHBD livers. TUNEL staining after 8-hour cold storage. In HTK group, in spite of preserved architecture, there appeared many TUNEL positive hepatocytes diffusely. Conversely, only a few TUNEL positive cells were noticed in UW group, despite hepatocytes showed marked shrinkage.[Conclusion] Instead of HBD livers, higher osmolarity of UW solution may be harmful in the preservation of NHBD livers. In NHBD graft, limitation of apoptosis during cold preservation did not necessarily contribute to maintaining a better graft viability. ETK solution seemed to have advantage, compared with HTK solution. However, ETK solution was enough to maintain NHBD graft viability. A better strategy in preservation method for salvaging NHBD livers is necessary. Less

[简介]非心脏跳动供体(NHBD)肝脏有望成为解决供体长期短缺的另一种移植物来源。无节制的NHBD移植物通常会遭受广泛的热缺血损伤，因此移植效果不佳。为了明确保存NHBD肝脏的意义，我们比较了目前可用的UW、HTK和ETK(ETK)三种溶液对NHBD肝脏保存后的损伤。[方法]Wistar大鼠在心脏骤停60或30分钟后切开膈肌，取肝。经门静脉注入尿激酶液溶解肝窦内血栓，室温放置10min。分别用80ml冷UW液、HTK液、ETK液冲洗肝脏，4℃保存8h。以未保存的HBD肝作为对照。我们使用隔离肝脏灌流模型评估移植物的存活能力。复温20分钟后，li…充氧Krebs Henseleit缓冲液灌流更多的VERS。进行60分钟的灌流。在灌流过程中监测门静脉压力和胆汁生成。测定灌流液中AST水平。[结果]注射尿激酶液后移植物存活率明显提高。ETK组门静脉压力低于UW组和HTK组。再灌流60min，ETK组胆汁生成量明显高于UW组和HTK组。ETK组与UW组比较，差异有统计学意义。然而，ETK组的胆汁产量仅为HBD肝脏的三分之一。ETK组灌流液中AST水平低于UW组和HTK组。UW组和ETK组仅在再灌流5min时差异有统计学意义。HE染色，UW组肝细胞明显缩小，肝窦间隙明显延长。另一方面，HTK组肝脏结构保存较好。相反，较高渗透压的UW液可能有利于NHBD肝损伤前肝细胞的损伤。冷藏8小时后进行TUNEL染色。HTK组尽管结构完整，但仍可见大量TUNEL阳性肝细胞弥漫分布。相反，UW组尽管肝细胞明显萎缩，但仅有少量TUNEL阳性细胞。[结论]较高渗透压的UW液可能不利于NHBD肝脏的保存。在NHBD移植物中，冷保存期间限制细胞凋亡并不一定有助于维持较好的移植物存活率。与HTK溶液相比，ETK溶液似乎具有优势。然而，ETK液足以维持NHBD移植物的活力。抢救NHBD肝脏的保存方法需要一种更好的策略。较少

项目成果

Non-heart-beating donor (NHBD)摘出肝のグラフト使用を目指して

旨在使用无心跳供体（NHBD）肝移植

• 发表时间: 2007
• 作者: Masunaga;S.;Sakurai;Y.;Nagata;K.;Suzuki;M.;Maruhashi;A.;Kinashi;Y.;Nagasawa;H.;Uto;Y.;Hori;H.;Ono;K;増永 慎一郎;菊地功;Isao Kikuchi;菊地 功
• 通讯作者: 菊地 功

Viability of liver graft from non-heart-beating donor

来自无心跳供体的肝移植物的活力

• 发表时间: 2007
• 作者: Masunaga;S.;Sakurai;Y.;Nagata;K.;Suzuki;M.;Maruhashi;A.;Kinashi;Y.;Nagasawa;H.;Uto;Y.;Hori;H.;Ono;K;増永 慎一郎;菊地功;Isao Kikuchi
• 通讯作者: Isao Kikuchi

Non-heart-beating donor(NHBD)摘出肝のグラフト使用を目指して

旨在使用无心跳供体（NHBD）肝移植

• 发表时间: 2007
• 作者: Masunaga;S.;Sakurai;Y.;Nagata;K.;Suzuki;M.;Maruhashi;A.;Kinashi;Y.;Nagasawa;H.;Uto;Y.;Hori;H.;Ono;K;増永 慎一郎;菊地功
• 通讯作者: 菊地功