1996 Fiscal Year Annual Research Report

肝類洞壁細胞を標的とした遺伝子治療による肝移植後肝不全の対策

Research Project

Project/Area Number	07670611
Research Institution	Saitama Medical School
Principal Investigator	持田智埼玉医科大学, 医学部, 講師 (20219968)
Co-Investigator(Kenkyū-buntansha)	藤原研司埼玉医科大学, 医学部, 教授 (80101088)
Keywords	肝移植 / 肝不全 / 肝類洞内皮細胞 / トロンボモジュリン / TFPI / VEGF
Research Abstract	申請者らは,肝類洞内皮細胞はトロンビンの不活性化に際して最も重要な抗凝固因子と考えられているトロンボモジュリンの発現が軽微であることを見いだした。更に,tissue factorによる血液凝固の開始を抑制するtissue factor pathway inhibitor(TFPI)が肝類洞内皮細胞には発現していないことも明らかにした。これら他臓器末梢血管と著しく異なる肝類洞の内皮細胞における特殊性は,同所性肝移植後肝不全の成立に関与していると考えられ,標的抗凝固療法としてはTFP1及びトロンボモジュリンを補給する方針を決定した。 TFPIはヘパリンと結合する性質を利用して,蛋白レベルでの標的療法が可能であることを明らかにした。組替え型TFPIをラットに静注すると血中から速やかに消失するが,抗TFPIポリクローナル抗体を用いた免疫電顕により類洞内皮細胞及びDisse腔内の肝細胞微縦毛膜に結合していることを証明した。この染色性はヘパリンナトリウムを静注すると減弱し,TFPIが血中で再び検出されることから同部のヘパリノイドに結合していると考えられた。トロンボモジュリンはcDNAを用いた遺伝子レベルでの標的療法の開発が進行中である。遺伝子導入に必要なトロンボモジュリンの全長cDNAをクローニングした。疎水性のポリビニルと親水性のマンノースを結合して水層内でミセルを作成し,マンノース受容体を利用して類洞内皮細胞にcDNAを導入する方法を確立した。また,移植肝では類洞内皮細胞が障害されているため,その再生を促進することも類洞内凝固の治療に際して有効と考えられる。申請者らは,類洞内皮細胞の再生にはVGEFを介した肝構成細胞の相互作用が重要な役割を担っていることを明らかにし,これを調節する治療法の端緒をも見いだした。

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] Mochida S,OhnoA,FujiwaraK.: "Pitfall in the inplication of intercellular adhesion molecule-1 expression on isolated hepatocytes." Hepatology. (in press).
[Publications] Inao,M,Mochida S,et al.: "Effects of neurotopic pyrimidine heterocyclic compound,MS-430 on cultured hepatic parenchymal and stellate cells." Life Sciences. (in press).
[Publications] Yamanobe,Mochida S,et al.: "Recombinant human tissue factor pathway inhibitor as a possible anticoagulant targeting hepatic sinusoidal walls." Thrombosis Res. (in press).
[Publications] Mochida S,Arai M,Ohno A,Fujiwara K.: "Bacterial translocation from gut to portal blood as a factor of hypercoagulopathy in the hepatic sinusoids following orthotopic liver transplanation in rats." Transplant Proc. 29. 874-875 (1997)
[Publications] Arai M,Mochida S,Ohno A,Fujiwara K.: "Blood coagulation in the hepatic sinusoids as a factor of liver injury following orthotopic liver transplantation in rats." Transplantation. 62. 1398-1401 (1996)
[Publications] Mochida S,Ishikawa K,et al.: "Increased expression of vascular endothelial growth factor and its receptors,flt-1 and KDR/flk-1,in regenerating rat liver" Biochem Biophys Res Commun. 226. 176-179 (1996)
[Publications] Mochida S,Ohno A,et al.: "Role of adhesion molecules in the development of massive hepatic necrosis in rats." Hepatology. 23. 320-328 (1996)
[Publications] Ohno A,Mochida S,Arai M,Fujiwara K.: "ICAM-1 expression in hepatocytes following dissociation of cell-to-cell contact in rats." Biochem Biophys Res Commun. 214. 1225-1231 (1995)
[Publications] Arai M,Mochida S,et al.: "Blood coagulation equilibrium in rat liver microcirculation as evaluated by endothelial cell thrombomodulin and macrophage tissue factor." Thromb Res. 80. 118-123 (1995)
[Publications] Mochida S,Ohno A,Arai M,Fujiwara K.: "Role of adhesion between activated macrophages and endothelial cells in the development of two types of massive hepatic necrosis in rats." J Gastroenterol Hepatol. 10. S38-S42 (1995)
[Publications] Fujiwara K,Mochida S,Ohno A,Arai M.: "Possible cause of primary graft nonfunction after orthotopic liver transplantation : a hypothesis with rat models." J Gastroenterol Hepatol. 10. S88-S91 (1995)
[Publications] Fujiwara K,Mochida S,et al.: "Use of Prostaglandin I2 analog in the treatment of massive hepatic necrosis associated with endothelial injury and diffuse sinusoidal tibrin deposition." Dig Dis Sci. 40. 41-47 (1995)
[Publications] Fujiwara K,Mochida S,et al.: "Excerpta Medica,International Congress Series 1086 Progress in Hepatology "Cellular and Molecular Biology"" Yamanaka M,et al.(eds),Elsevier Science B.V.,Amsterdam, 144 (1995)

1996 Fiscal Year Annual Research Report

肝類洞壁細胞を標的とした遺伝子治療による肝移植後肝不全の対策

Principal Investigator

持田 智 埼玉医科大学, 医学部, 講師 (20219968)

Research Products

[Publications] Mochida S,OhnoA,FujiwaraK.: "Pitfall in the inplication of intercellular adhesion molecule-1 expression on isolated hepatocytes." Hepatology. (in press).

[Publications] Inao,M,Mochida S,et al.: "Effects of neurotopic pyrimidine heterocyclic compound,MS-430 on cultured hepatic parenchymal and stellate cells." Life Sciences. (in press).

[Publications] Yamanobe,Mochida S,et al.: "Recombinant human tissue factor pathway inhibitor as a possible anticoagulant targeting hepatic sinusoidal walls." Thrombosis Res. (in press).

[Publications] Mochida S,Arai M,Ohno A,Fujiwara K.: "Bacterial translocation from gut to portal blood as a factor of hypercoagulopathy in the hepatic sinusoids following orthotopic liver transplanation in rats." Transplant Proc. 29. 874-875 (1997)

[Publications] Arai M,Mochida S,Ohno A,Fujiwara K.: "Blood coagulation in the hepatic sinusoids as a factor of liver injury following orthotopic liver transplantation in rats." Transplantation. 62. 1398-1401 (1996)

[Publications] Mochida S,Ishikawa K,et al.: "Increased expression of vascular endothelial growth factor and its receptors,flt-1 and KDR/flk-1,in regenerating rat liver" Biochem Biophys Res Commun. 226. 176-179 (1996)

[Publications] Mochida S,Ohno A,et al.: "Role of adhesion molecules in the development of massive hepatic necrosis in rats." Hepatology. 23. 320-328 (1996)

[Publications] Ohno A,Mochida S,Arai M,Fujiwara K.: "ICAM-1 expression in hepatocytes following dissociation of cell-to-cell contact in rats." Biochem Biophys Res Commun. 214. 1225-1231 (1995)

[Publications] Arai M,Mochida S,et al.: "Blood coagulation equilibrium in rat liver microcirculation as evaluated by endothelial cell thrombomodulin and macrophage tissue factor." Thromb Res. 80. 118-123 (1995)

[Publications] Mochida S,Ohno A,Arai M,Fujiwara K.: "Role of adhesion between activated macrophages and endothelial cells in the development of two types of massive hepatic necrosis in rats." J Gastroenterol Hepatol. 10. S38-S42 (1995)

[Publications] Fujiwara K,Mochida S,Ohno A,Arai M.: "Possible cause of primary graft nonfunction after orthotopic liver transplantation : a hypothesis with rat models." J Gastroenterol Hepatol. 10. S88-S91 (1995)

[Publications] Fujiwara K,Mochida S,et al.: "Use of Prostaglandin I2 analog in the treatment of massive hepatic necrosis associated with endothelial injury and diffuse sinusoidal tibrin deposition." Dig Dis Sci. 40. 41-47 (1995)

[Publications] Fujiwara K,Mochida S,et al.: "Excerpta Medica,International Congress Series 1086 Progress in Hepatology "Cellular and Molecular Biology"" Yamanaka M,et al.(eds),Elsevier Science B.V.,Amsterdam, 144 (1995)

持田智埼玉医科大学, 医学部, 講師 (20219968)