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目的建立一种稳定的小鼠肝脏持续低灌注模型,并在此基础上研究肝脏持续低灌注对小鼠肝脏热缺血再灌注损伤的影响。方法选用6~8周龄C57BL/6小鼠建模,将门静脉缩窄至1mL注射器针头直径,门静脉缩窄后3d、7d、14d和21d行肝功能及肝脏组织病理学检测;选用稳定的模型小鼠行70%缺血再灌注手术,再灌注3h、24h、48h后行肝功能及肝脏组织病理学检测。对照组采用正常C57BL/6小鼠行缺血再灌注手术。结果小鼠门静脉缩窄术后,丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)均有不同程度的升高,在7d时达到高峰[ALT:(60.8±6.2)U/L vs(25.5±2.8)U/L,P<0.001;AST:(74.9±6.1)U/L vs(39.1±3.2)U/L,P<0.001),同时H-E染色显示7d时肝细胞损伤最重,并且有较多炎细胞浸润;在21d时,ALT基本恢复正常水平(P>0.05),而AST仍高于正常水平(P=0.03)。低灌注处理7d的小鼠进行缺血再灌注手术后,肝酶和组织病理学检查显示肝细胞损伤较对照组显著加重,肝酶在再灌注3h达到高峰[ALT:(8 217.0±1 111.8)U/L vs(5 597.4±1 015.3)U/L,P=0.004;AST:(8 548.2±1 155.4)U/L vs(5 765.4±956.9)U/L,P=0.003];再灌注48h时,对照组小鼠ALT和AST均恢复正常,而经过低灌注处理的小鼠肝酶仍高于对照组[ALT:(608.8±442.9)U/L vs(47.4±20.1)U/L,P=0.008;AST:(861.8±442.8)U/L vs(70.8±68.3)U/L,P=0.008)。结论成功建立了稳定的小鼠肝脏持续低灌注模型,经持续低灌注处理后的肝脏对热缺血再灌注损伤的耐受能力显著降低,这在一定程度上能够模拟临床上心死亡器官捐献供肝的状况。
Objective To establish a stable low-perfusion model of liver in mice and to study the effect of sustained low perfusion of liver on hepatic ischemia-reperfusion injury in mice. Methods C57BL / 6 mice aged 6 to 8 weeks were selected and the portal vein was narrowed to the diameter of 1 mL syringes. The liver function and liver histopathology were examined at 3d, 7d, 14d and 21d after portal vein constriction. Stable model The mice were subjected to ischemia-reperfusion surgery 70% and reperfusion 3h, 24h, 48h after liver function and liver histopathological examination. The control group was given normal C57BL / 6 mice for ischemia-reperfusion. Results After ameliorating portal vein, the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) increased to some extent and reached the peak at 7d [ALT: (60.8 ± 6.2) U / L vs (25.5 ± 2.8) U / L, P <0.001; AST: (74.9 ± 6.1) U / L vs (39.1 ± 3.2) U / L, P <0.001) (P> 0.05), while the level of AST was still higher than the normal level (P = 0.03). After ischemia-reperfusion, the liver enzymes and histopathological examination showed that liver injury was significantly aggravated in mice treated with low perfusion for 7 days, and reached the peak at 3 hours after reperfusion (ALT: (8 217.0 ± 1 111.8) U / L vs 5 597.4 ± 1 015.3 U / L, P = 0.004; AST: (8 548.2 ± 1 155.4) U / L vs (5765.4 ± 956.9) U / L, P = 0.003] , ALT and AST returned to normal in the control mice, while the liver enzymes in the low perfusion mice were still higher than those in the control group [ALT: (608.8 ± 442.9) U / L vs (47.4 ± 20.1) U / L, P = 0.008; AST: (861.8 ± 442.8) U / L vs (70.8 ± 68.3) U / L, P = 0.008). Conclusion The stable model of persistent liver hypoperfusion in mice has been successfully established. The tolerance of the liver to warm ischemia-reperfusion injury after continuous hypoperfusion is significantly reduced, which to some extent can simulate the clinical donation of cardiac death organs Liver condition.