特异性敲除β-连环蛋白基因对小鼠肝缺血再灌注损伤的影响  

Conditional deletion of β-catenin reveals its role in liver ischemia-reperfusion injury

作　　者：周文华 霍守俊[2] 孔凡志[2] 周联明 张学利 

机构地区：[1]上海奉贤区中心医院普外科,201400 [2]苏州大学医学院

出　　处：《中华实验外科杂志》2011年第6期926-928,I0003,共4页Chinese Journal of Experimental Surgery

基　　金：基金项目：上海市科委启动基金资助项目（8411963700）

摘　　要：目的通过构建特异性敲除小鼠模型观察β-连环蛋白（13-catenin）基因在小鼠肝缺血再灌注损伤中的作用。方法应用Cre—lox系统构建肝细胞特异性敲除B—catenin基因小鼠模型，90min半肝血流阻断制造肝脏缺血再灌注小鼠模型，分为B—catenin基因敲除组及对照组，再灌注后0…3620h测血清谷丙转氨酶（ATJT）、谷草转氨酶（AST）及其肝脏组织学改变。结果肝细胞特异性敲除p—catenin基因小鼠基因表型为：ctnnb纯合（10xP／loxP）并且Alb—Cre阳性表达，Westernblot方法证实其β-catenin蛋白表达丰度明显低于对照组。再灌注3、6、20h敲除组小鼠血清ALT水平分别为282、405及128U／ml；对照组分别为221、295及101U／ml。敲除组AST水平分别为603、805及396U／ml；对照组分别为421、398及336U／ml，各时间点两组间差异均有统计学意义（P〈0．05）。其肝细胞坏死、窦状隙充血以及炎细胞浸润程度明显重于对照组。结论B—catenin对肝脏缺血再灌注引起的肝损伤起保护作用。Objective To study the role of β-catenin in liver ischemia-reperfusion injury used a conditional knockout approach to delete β-catenin in the liver. Methods We adapted the Cre-lox recombination system to create the β-catenin conditional knockout （KO） mice and liver ischemia-reperfusion injury were made by 90min hemiliver blood supply block. Experimental animal were randomized into two groups： β-catenin conditional knockout group （KO group） and control group （C group）. 0,3,6,20 h after reperfusion, serum samples were used for measurement of serum alanine transaminase （ALT） and aspartate aminotransferase （AST） ,liver sections were subjected to hematoxylin-eosin staining. Results Genotyping identified that β-catenin KO mice showed ctnnb homozygosis （ -/- ） and Alb-Cre positive. Western blotting analysis showed an obviously lower expression of β-catenin in KO group. Three, 6,20 h after reperfusion, KO group ALT level were 282,405 and 128 U/ml. C group were 221,295 and 101 U/ml. KO group AST level were 603,805 and 396 U/ml, C group were 421,398 and 336 U/ml. Hematoxylin and Eosin （HE） staining showed significant sinusoid inflation and congestion, extensive acidophilic necrosis ,focal hemorrhages, hepatic lobula structure disorder and slightly lymphocyte inflammatory cells infiltration in KO group. In C group, HE staining show slightly sinusoid inflation and congestion,limited acidophilic necrosis and focal hemorrhages,hepatic lobula structure nearly integrity, slightly lymphocyte inflammatory cells infiltration. Conclusion β-catenin may play a protect role in liver ischemia-reperfusion injury.

关 键 词：Β-连环蛋白 缺血再灌注损伤 基因敲除 肝脏 小鼠 

分 类 号：R285.5[医药卫生—中药学]