TNF-α a antisense transfer improves ischemia / reperfusion injury of the hepatic graft

TNF-α反义转移改善肝移植物的缺血/再灌注损伤

• 批准号: 08407031
• 负责人: SUGIMACHI Keizo
• 金额: $ 25.66万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08407031/
• 关键词: cold ischemia; reperfusion injury; liver transplantation; HVJ-CL; TNF-α antisense; Kupffer cells; oligodeoxynucleotides; IL-1β; TNF-α; 冷虚再潅流障害; 遺伝子導入; TNFα; NF-κB; decov; 肝臓移植; 冷虚血再潅流障害; NF-κ B decoy; 冷虚血再潅流傷害; Apoptosis

Cold ischemia/reperfusion injury of the hepatic graft, an unsolved problem in liver transplantation that has been attributed to the release of inflammatory cytokines from activated Kupffer cells. We recently designed a novel vehicle composed of cationic liposomes fused with hemagglutinating virus of Japan (HVJ), namely HVJ-cationic liposomes (HVJ-CL), and using this vector system, we assessed the cytotoxicity, efficiency of Fluorescein isothiocyanate (FITC)-labeled oligodeoxynucleotides (FITC-ODNs) delivery to Kupffer cells and effects of TNF-α antisense delivery against ischemia/reperfusion injury in a rodent hepatic graft. FITC signals were recognized in over 95% of ED2-positive Kupffer cells of transplanted liver 4hrs after FITC-ODNs transfer. The transfer efficiency (18.5 ± 3.1%) using HVJ-CL applied at one-eighth the amount of HVJ and lipids of conventional HVJ-liposomes (HVJ-L) was much higher than that achieved by HVJ-L (10.4 ± 1.2%, p<0.01), while there was no signincant difference in the vector-induced release of cytosolic enzymes from hepatocytes into the blood circulation between HVJ-CL and HVJ-L groups. Treatment with TNF-α antisense transfer significantly suppressed the release of cytosolic enzymes from hepatocytes, the serum levels of TNF-α and immunohistochemical expression of TNF-α by Kupffer cells 4hrs after liver transplantation. This HVJ-CL method provides a safe and effective ODNs delivery to Kupffer cells of hepatic grafts in rats

移植肝冷缺血/再灌注损伤是肝移植中尚未解决的问题，其原因是激活的枯否细胞释放炎性细胞因子。我们设计了一种新型的脂质体-日本血凝病毒（HVJ）-阳离子脂质体（HVJ-CL）载体系统，并利用该载体系统，研究了异硫氰酸荧光素（FITC）标记的寡脱氧核苷酸（FITC-ODNs）对枯否细胞的细胞毒性、转染效率以及TNF-α反义核酸对移植肝缺血/再灌注损伤的影响。FITC-ODNs转染后4 h，移植肝中95%以上的ED 2阳性枯否细胞可识别FITC信号。使用以常规HVJ-脂质体（HVJ-L）的HVJ和脂质的量的八分之一施加的HVJ-CL的转移效率（18.5 ± 3.1%）远高于HVJ-L实现的转移效率（10.4 ± 1.2%，p<0.01），而在HVJ-CL和HVJ-L组之间，载体诱导的细胞溶质酶从肝细胞释放到血液循环中没有显著差异。TNF-α反义基因转染可显著抑制肝移植后4 h肝细胞胞浆酶的释放、血清TNF-α水平和枯否细胞TNF-α的免疫组化表达。HVJ-CL法是一种安全有效的将ODNs导入大鼠移植肝Kupffer细胞的方法

项目成果

Suehiro T., Boros P., Emre S., Sheiner PA., Guy S., Schwartz Me., Miller CM.: "7. Donor des-gamma-carboxy prothrombin positivity is a risk factor for poor early glaft function in liver transplantation."Transplant Int. 11. 143-146 (1998)

Suehiro T.、Boros P.、Emre S.、Sheiner PA.、Guy S.、Schwartz Me.、Miller CM.：“7. 供体 des-gamma-羧基凝血酶原阳性是早期肝移植功能不良的危险因素

池上 徹、他: "Experiment study of the effects of deletion variant hepatocyte growth factor on hepatic ischemia-reperfusion injury."British journal of Surgery. 87. 59-64 (1999)

Toru Ikegami 等人：“删除变异型肝细胞生长因子对肝脏缺血再灌注损伤影响的实验研究。”英国外科杂志 87. 59-64 (1999)。

池上 徹、他: "Deletion variant of hepatocyte growth factor prolongs allograft survival after liver transplantation in rats."Surgery. 125. 602-607 (1999)

Toru Ikegami 等人：“肝细胞生长因子的缺失变异可延长大鼠肝移植后同种异体移植物的存活率。”Surgery。125. 602-607 (1999)

橋元 宏治、他: "Beneficial effect of FR167653 on cold ischemia/reperfusion injury in rat liver transplantation."Transplantation. 70. 1318-1322 (2000)

Koji Hashimoto 等人：“FR167653 对大鼠肝移植中冷缺血/再灌注损伤的有益作用。”移植。 70. 1318-1322 (2000)

Nishizaki T., Okano S., Uchiyama H., Ikegami T., Hashimoto K., Minagawa R., Ota R., Nomoto K., Yanaga K., Sugimachi K.: "6. Intraoperative hyaluronic acid clearance predicts postoperative graft function in living related liver transplantation."Int Surg..

Nishizaki T.、Okano S.、Uchiyama H.、Ikegami T.、Hashimoto K.、Minakawa R.、Ota R.、Nomoto K.、Yanaga K.、Sugimachi K.：“6. 术中透明质酸清除率预测术后移植物