Analysis of hepatic microcirculation during liver ischemia and reperfusion using in vivo microscopy

使用体内显微镜分析肝脏缺血和再灌注期间的肝脏微循环

• 批准号: 08671461
• 负责人: KAWANO Katsunori
• 金额: $ 1.28万
• 依托单位: Oita Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671461/
• 关键词: Hepatic microcirculation; In vivo microscopy; Liver ischemia; Liver transplantation; Neutrophil adhesion

BackgroundThe aim of the studies were to analyze, using the in vivo microscopic technique, the hepatic microcirculation during liver ischeniia and reperfusion, which was unavoidable in clinical liver transplantation and hepatic resection. In order to develop the new therapeutic strategies, inhibition of neutrophil adherence to vascular endothelia was attempted.Results1. Neutrophil adherence to hepatic vascular endothelia In a rat liver transplantation model, the liver graft was harvested and stored for 18 hours at 4ﾟC in UW solution. Then the hepatic microcirculation was investigated using in vivo microscopy following implantation. Sinusoidal perfusion rates were significantly lower in stored and transplanted liver (64%) compared with non-transplanted control liver (99%). Neutrophil adhesion to both sinusoidal endothelium (328/mm^2) and postsinusoidal venules (PSVs) were remarkably increased 24 hours following reperfusion of the liver graft.2. Amelioration of ischemia/reperfusion injur … More y using FK5O6 and anti-neutrophil adhesion molecule antibodyAn anti-CD 11b/CD 18 monoclonal antibody (clone 1B6, Repligen Co., Ltd.) was intravenously administered 30 minutes before and 6 hours after implantation of the 18-hour stored liver. The antibody therapy significantly improved graft survival (100% in the treated animals and 50% in the control) and reduced reperfusion injury to the liver as estimated serum ALT levels at 24 hours following transplantation. This was associated with significant inhibition of neutrophil adhesion to the sinusoidal endothelium (81/mm^2). An iMmunosuppressant, FK506, had similar effect ; prevented oxidative hepatic injury following cold preservation and transplantation by reducing the number of neutrophils permanently adhering to the sinusoids (15.4/mm^2 in FK-treated animals and 67.9/mm^2 in the control at 60 minutes following).ConclusionThese studies clearly demonstrated the causative role of neutrophils in hepatic injury following ischemia and reperfusion and suggested the possible clinical use of FK506 and antibody against neutrophil adhesion for the therapy. Less

本研究的目的是利用活体显微镜技术分析临床肝移植和肝切除术中不可避免的肝缺血再灌流过程中肝脏微循环的变化。为了开发新的治疗策略，我们尝试了抑制中性粒细胞与血管内皮细胞的黏附。在大鼠肝移植模型中，取肝组织，在4ﾟC的UW溶液中保存18h。在体内显微镜下观察植入后肝脏微循环的变化。保存肝和移植肝的肝窦灌注率(%)明显低于未移植对照肝(99%)。结论：1.移植肝再灌流24小时后，中性粒细胞与肝窦内皮细胞(328个/mm~2)和肝窦后小静脉(PSV)的黏附显著增加。改善缺血再灌注损伤…的实验研究多用FK5O6和抗中性粒细胞黏附分子抗体抗CD11b/CD18单抗(克隆1B6，Repligen Co.，Ltd.)分别在植入18h保存肝脏前30分钟和植入后6小时静脉给药。根据移植后24小时血清ALT水平的估计，抗体治疗显著提高了移植物存活率(处理组为100%，对照组为50%)，并减少了对肝脏的再灌注损伤。这与中性粒细胞与肝窦内皮细胞黏附的显著抑制有关(81/mm^2)。免疫抑制剂FK506具有类似的作用；通过减少中性粒细胞永久贴壁肝窦的数量(FK组为15.4个/mm2，对照组为67.9个/mm2)，预防冷保存和移植后的氧化性肝损伤。结论本研究明确了中性粒细胞在肝缺血再灌注损伤中的致病作用，提示FK506和抗中性粒细胞黏附抗体可能用于临床治疗。较少

项目成果

Kawano K,et al.: "Evidence that both cyclosporin and azathioprine prevent warm ischemia reperfusion injury to the rat liver." Transplant International. 6.6. 330-336 (1993)

Kawano K 等人：“有证据表明环孢素和硫唑嘌呤均可预防大鼠肝脏的热缺血再灌注损伤。”

Kawano K,et al.: "Protective effect of FK506 on hepatic injury following cold ischemic preservation and transplantation:influence on hepatic microcirculation." Transplantation Proceedings. 27.1. 362-363 (1996)

Kawano K,et al.：“FK506 对冷缺血保存和移植后肝损伤的保护作用：对肝脏微循环的影响”。

Kawano K,et al.: "FK506 ameliorates normothermic liver ischemia in rats by suppressing production of tumor necrosis factor." Transplant International. 5.suppl 1. 665-669 (1992)

Kawano K 等人：“FK506 通过抑制肿瘤坏死因子的产生来改善大鼠的常温肝缺血。”

Kawano K,et al.: "FK506 reduces oxidative hepatic injury following cold ischemic preservation and transplantation" Transplantation Proceedings. 28.3. 1902-1903 (1996)

Kawano K 等人：“FK506 减少冷缺血保存和移植后的氧化性肝损伤”移植论文集。

Kawano K,et al.: "Protective effect of FK506 on hepatic injury following cold ischemic preservation and transplantation : influence on hepatic microcirculation." Transplantation Proceedings. 27.1. 362-363 (1996)

Kawano K 等人：“FK506 对冷缺血保存和移植后肝损伤的保护作用：对肝脏微循环的影响”。