解耦联蛋白-2基因敲除减轻高脂饮食诱导小鼠脂肪肝缺血再灌注损伤

目的 观察解耦联蛋白-2(UCP-2)基因在脂肪肝缺血再灌注损伤中的作用,探讨以UCP-2为靶点的脂肪供肝预处理新途径.方法 实验动物分为3组:实验组(A组):UCP-2基因敲除小鼠,给予高脂饮食(n=20);空白对照组(B组):野生型同系小鼠,给予正常饮食(n=20);阴性对照组(C组):野生型同系小鼠,给予高脂饮食(n=20).喂养6个月,建屯小鼠脂肪肝模型.各组小鼠阻断门静脉缺血15 min,再灌注24 h后处死,检测肝组织中UCP-2基因表达及三磷酸腺苷(ATP)、活性氧族(ROS)、乳酸脱氢酶(LDH)水平;并行血清丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)检测及肝组织病理学检测.结果 高脂饮食喂养6个月后成功建立小鼠脂肪肝模型;Western blot证实A组小鼠UCP-2无表达,C组表达明显高于B组;A组ALT、AST、ATP、ROS、LDH的定量值为:(55.33±5.51)、(128.33±7.02)U/L、(28.00±2.00)nmol/L、(165.33±7.09)、(1176.00±22.27)U/pmt;B组上述指标为(25.00±4.58)、(85.33±4.51)U/L、(24.33±4.16)nmol/L、(147.33±7.51)、(707.33±31.64)U/prot;C组为(142.67±13.01)、(220.67±7.02)U/L、(8.67±1.53)nmol/L、(65.00±5.00)、(1748.33±42.52)U/prot,A组和C组差异有统计学意义(P<0.05);病理检测表明A组损伤轻于C组.结论 抑制解耦联蛋白-2基因表达能在减轻小鼠脂肪肝急性损伤.

UCP-2 gene knock-out alleviates ischemia/reperfusion-induced injury of high fat diet induced fatty liver

Objective To investigate the role of uncoupling protein-2 (UCP-2) in ischemia-reperfusion-(P<0.05) induced injury and explore a new target for fatty donor hver pretreatment. Methods The animals were divided into 3 groups: experimental group (group A) : UCP2 gene knock-down mice, feeding on high-fat diet; blank control group (group B) : homologous series wild type mice, feeding on normal diet; na-gative control group (group C) : homologous series wild type mice, feeding on high-fat diet. Six months later, the mice in every group were sacrificed. The levels of adenosine-triphosphate (ATP), reactive oxygen species (ROS), lactate dehydrogenase (LDH) in hver tissues, and alanine tronsaminase (ALT), and asparate amin-otransferase (AST) in serum were determed. The pathologic changes were also examined. Results After feed-ing on high fat diet for 6 mouths, the model of fatty liver in mice was established. Western blotting con-firmed that the expression of UCP-2 mRNA was inhibited completely, and the expression level in group C was higher than in group B. The ALT, AST, ATP, ROS and LDH levels in group A were (55.33± 5.51), (128.33±7.02) U/L, (28.00±2.00) nmol/L, (165.33±7.09), (1176.00±22.27) U/prot, those in group B were (25.00±4.58), (85.33±4.51) U/L, (24.33±4.16) nmol/L, (147.33± 7.51), and (707.33±31.64) U/prot, and those in group C were (142.67±13.01), (220.67±7.02) U/L, (8.67±1.53 ) nmol/L, (65.00±5.00), (1748.33±42.52) U/prot, respectively. There was significant difference between group A and group C. Pathologic examination revealed that in group A, the ischemia-reperfusion-induced injury was milder than in group C. Conclusion Inhibiting the expression of UCP-2 attenuates the I/R injury of mouse fatty liver.