過小グラフトを用いた肝移植における、肝機能障害の機序解明と制御法の確立

阐明小尺寸肝移植中肝功能障碍的机制及控制方法的建立

• 批准号: 17790890
• 负责人: 丸橋 繁
• 金额: $ 2.24万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17790890/
• 关键词: 肝再生; ラット; 肝切除; mRNA; HGF; IL-6; 肝切除後肝再生; サイトカイン; MRP-2; 過小グラフト; 遺伝子導入

本研究では、(1)大量肝切除後あるいは過小グラフト肝移植後は、肝再生が急激に起こり、本来の生命維持に必要な肝代謝が抑制され胆汁欝滞・肝機能障害をもたらす、(2)肝代謝を促進することにより胆汁欝滞を軽減し肝機能障害を抑制することができる、という仮説をたて、研究を開始した。ラット(WSラットRT1k)7週齢、体重280〜320gを実験に用いた。麻酔はペントバルビタールを使用した。まずラット70%肝切除モデルの検討を行った。開腹ののち、Higgins変法を用いて70%肝切除を行った。術前、及び術後6、12、18、24時間後に安楽死させ下大静脈より採血、残肝の摘出を行なった。血液はヘパリン及びEDTA2Naと混合し、遠沈後血漿、血清を採取し検査まで-20℃で保存し、肝組織は二分し一片をsnap frozenの後-80℃で保存、一片を10%緩衝ホルマリンで保存した。末梢血中のHGF濃度は肝切除後6時間で前値の10倍ほどに上昇し術後24時間まで維持された。肝組織中のHGFは、術後6時間以降で前値の倍ほどに上昇した。また、この肝組織中のHGFは肝切除前に比べ高率に活性化されており、大量肝切除後の肝再生にはHGFの活性化が関与していることが結論された。肝組織中IL-6mRNAは術後6時間で急激の上昇し術後12時間以降は減少した。一方TNFαmRNAは時間とともに増加する傾向にあった。また、ラット大量肝切除モデルを用いてMRP-2発現をmRNA、蛋白レベルで検討し、MRP-2強制発現のため肝細胞への遺伝子導入を試みた。本研究の結果から、ラット肝臓の大量肝切除後には肝組織内のIL-6、TNFαのmRNAの発現が急増し、一方でHGFの肝組織内の増加および活性化が起こること、これにより肝再生が体系的に起こることが示された。

In this study, (1) a large number of patients after hepatectomy were diagnosed after liver transplantation, acute attack of liver regeneration, inhibition of bile stagnation and liver failure after hepatectomy. in this study, (1) after massive hepatectomy, (1) a large number of patients after hepatectomy, liver transplantation, acute liver regeneration, essential life maintenance, inhibition of bile stagnation, (2) promotion of bile stagnation, liver dysfunction, liver dysfunction, liver failure, liver failure, liver failure The research begins and begins. The body weight is 280g and 320g, and the body weight is 280g. The body weight is 280g and 320g. I don't know how to use it. A 70% hepatectomy was performed. Laparoscopic hepatectomy and Higgins hepatectomy were performed. Before surgery and 6, 12, 18 and 24 hours after surgery, the blood and remnant liver were removed during tranquilization during euthanasia. The blood samples were collected and stored at-20 ℃. The liver tissue was stored at-80 ℃ after snap frozen, and the liver tissue was stored at-80 ℃. The concentration of HGF in peripheral blood was 10 times higher than that before hepatectomy and maintained at 24 hours after hepatectomy. HGF was detected in the liver tissue during 6 hours after operation, and the serum levels were doubled before and after 6 hours. The positive rate of HGF in liver tissue before hepatectomy was higher than that before hepatectomy, and that of liver regeneration after massive hepatectomy was significantly higher than that before hepatectomy (P < 0.01). The ratio of HGF in liver tissue before hepatectomy was higher than that before hepatectomy. The IL-6mRNA in the liver tissue was reduced at 6 hours after shock and 12 hours after shock. One side of the tumor necrosis factor alpha mRNA in time to increase the number of patients in the hospital. After massive hepatectomy, MRP-2 was used to detect the presence of mRNA, protein, and MRP-2 to detect the presence of liver cancer. The results of this study showed that after massive hepatectomy, IL-6 and TNF α mRNA were found in the liver tissue of the liver group, and the activation of the liver tissue was activated in one side of the HGF liver tissue, and the liver regeneration system of the liver regeneration system was significantly affected.

项目成果

Alpha-fetoprotein mRNA Detection in Peripheral Blood for Prediction of Hepatocellular Carcinoma Recurrence After Liver Transplantation.

外周血中甲胎蛋白 mRNA 检测可预测肝移植后肝细胞癌复发。

• 发表时间: 2006
• 期刊: Transplant Proc 38(10)
• 作者: Marubashi S.;Dono K.;Sugita Y.;Asaoka T.;Hama N.;Gotoh K.;Miyamoto A.;Takeda Y.;Nagano H.;Umeshita K.;Monden M.
• 通讯作者: Monden M.

Liver transplantation for hepatitis C.

肝移植治疗丙型肝炎。

• 发表时间: 2006
• 期刊: J Hepatobiliary Pancreat Surg 13
• 作者: Marubashi S;Dono K.;Miymoto A.;Takeda Y.;Nagano H.;Umshita K.;Monden M.
• 通讯作者: Monden M.