小鼠原位肝移植模型及移植肝24h冷缺血再灌注模型的建立

Construction of orthotopic liver transplantation model in mouse and graft 24 hours cold ischemia-reperfusion injury model

目的探讨单人袖套法建立小鼠原位不重建动脉肝移植模型的效果,初步研究移植肝24 h冷缺血再灌注损伤模型的构建。方法术者接受单人袖套法建立小鼠原位不重建动脉肝移植模型的操作训练,每个训练周期完成10对小鼠的操作,共进行6个训练周期。分析每个训练周期内受体小鼠生存率以及受体小鼠生存率与术中无肝期时间的关系。3只小鼠接受冷保存24 h的供肝,以建立移植肝24 h缺血再灌注模型,术后6 h检测小鼠血清ALT、AST水平并对移植肝组织进行Suzuki评分。结果术者经过训练后,第6个训练周期内受体小鼠7,14,100 d的生存率分别为100%,90%,60%。术中无肝期时间≤20 min组与20 min<术中无肝期时间≤24 min组术后100 d生存率分别为56%和22%,差异有统计学意义(χ2=5.519,P<0.05)。3只接受24 h冷保存供肝的受体小鼠血清ALT、AST中位数分别为12 350 U/L、9 870 U/L,移植肝Suzuki评分为(10.6±0.6)分,提示移植肝出现了重度的组织学损伤。结论单人袖套法建立小鼠原位不重建动脉肝移植模型技术简单易学,能够满足研究对小鼠的存活需求。本研究建立的移植肝24 h冷缺血再灌注模型出现了损伤改变,能够满足后期研究需求。

Objective To construct a mouse orthotopic liver transplantation model, and graft 24 hours cold ischemia-reperfusion injury model. Methods Researchers accepted six operation training cycle to construct a mouse orthotopie liver transplantation model without artery reconstruction by cuff technique,each training cycle to complete 10 pairs of mice. The survival rate in each training period and the relationship between survival rate and anhepatie phase were analyzed. Grafts 24 hours cold ischemia-reperfusion injury model were established by cold preservation （3 cases ）, then the injury of grafts was estimated. Results After training, the 7, 14 and 100 days survival rate was 100% , 90% , 60% respectively in the sixth training period. Compared the groups with a longer time of anhepatic phase, the 100 days survival rate was higher in the group with anhepatic phase less than 20 minutes. In the graft 24 hours cold ischemia-reperfusion injury model, the median serum level of ALT and AST in recipient mice at 6 hours after reperfusion was 12 350 U/L, 9 870 U/L respectively. Suzuki score of grafts was 10.6 ±0. 6 that indicated a severe injury in histology. Conclusion Constructing the mouse orthotopic liver transplantation model by cuff technique was simple comparatively, graft 24 hours cold ischemia-reperfusion model was useful for the research.