冬眠誘導因子を用いた移植肝臓保存時間の延長に関する実験的研究

利用冬眠诱导因子延长移植肝保存时间的实验研究

• 批准号: 13770684
• 负责人: 川口 千晶
• 金额: $ 0.9万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13770684/
• 关键词: 肝臓移植; ATP; 冬眠誘導因子; 活性酸素; 肝保存; 類洞壁細胞; 肝細胞

Lewisラットの肝細胞、肝類洞壁細胞の単純浸漬冷保存下でのviabilityと再酸素化、復温時のviabilityを冬眠誘導因子投与の相違により比較した。1群:DADLE1mg/kg体重を全く投与しない群2群:DADLE1mg/kg体重を肝臓摘出24時間前と肝臓摘出直前に陰茎静脈より投与する群3群:DADLE1mg/kg体重を肝臓摘出直前に門脈から投与する群冷保存下と再酸素化での細胞内ATP量測定形態学的変化、・麻酔下にLewisラット(雄性8週令)の肝臓を潅流後、摘出し、肝類洞壁細胞、肝細胞を分離し細胞培養用wellにて4℃のUW液内で保存し。肝細胞、肝類洞壁細胞の細胞内ATP量、形態学的変化を、4℃のUW液にて保存しO,12,24,48,72時間保存後とその後に37℃で酸素化したKrebs Hensleit Bufferで30分培養させた後に測定、検討した。・麻酔下にLewisラット(雄性8週令)の肝臓を潅流後、摘出し、4℃のUW液にて保存する。保存後、24時間、72時間後に生体微小循環連続観察記録装置にマウントし酸素化したKrebs-Hansleit bicarbonate bufferにて潅流(frow rate3ml/min/kg)し、Dichlorofluorescein diacetate(DCFH)5μmol/l投与と、Propidium iodide(Pl)1μmol/l投与後の蛍光画像解析により活性酸素量測定と障害細胞同定を行った。結果:形態学的変化は3群間に差異は認めなかった。細胞内ATP量は、肝細胞のそれが2群で良好に保持されていた。再酸素化後の障害では、3群間に差異は認められなかった。

Comparison of Viability of Liver Cells and Liver Wall Cells in Pure Immersion and Cold Storage with Viability of Reacidification and Rewarming and Hibernation Induction Factor Group 1:DADLE1mg/kg body weight was administered 24 hours before liver harvesting and before penile vein harvesting Group 3:DADLE1mg/kg body weight was administered 24 hours before liver harvesting and before penile vein harvesting Group 3:DADLE1mg/kg body weight was administered 24 hours before liver harvesting and before portal vein harvesting Group 3:DADLE1mg/kg body weight was administered 24 hours before liver harvesting and before penile vein harvesting Group 3: DADLE 1 mg/kg body weight was administered 24 hours before liver harvesting Group 4:DADLE1mg/kg body weight Group 4: DADLE 1 Changes in intracellular ATP content and morphology of hepatocytes and hepatoid wall cells after preservation in UW solution at 4℃ for 0,12,24,48,72 hours and then in Krebs Hensleit Buffer at 37℃ for 30 minutes were measured and examined. The livers of Lewis rats (male 8 weeks old) were harvested and stored in UW solution at 4℃. After preservation, 24 hours and 72 hours, the micro-circulation of living organisms was continuously observed. The flow rate (frow rate3ml/min/kg) in Krebs-Hansleit bicarbonate buffer, Dchlorofluorene acetate (DCFH)5μmol/l and Propidium iodide(PI)1μmol/l were measured. Results: Morphological differences among the three groups were identified. Intracellular ATP content was well maintained in both groups of hepatocytes. After re-acidification, the difference between the three groups is recognized.