多功能蛋白APE1/Ref1在肝衰竭大鼠肝细胞不同部位的表达意义  

Expression of multifunctional protein APE1/ Ref1 in different parts of liver cells in rats with acute-on-chronic liver failure

作　　者：刁建新[1] 代欢[1] 李海叶[1] 杨运高[1] 

机构地区：[1]南方医科大学中医药学院,广州市510515

出　　处：《实用肝脏病杂志》2016年第2期151-155,共5页Journal of Practical Hepatology

基　　金：国家自然科学基金资助项目(编号:81173262)

摘　　要：目的研究多功能蛋白酶脱嘌呤/脱嘧啶核酸内切酶(APE1/Ref1)在慢加急性肝衰竭大鼠肝细胞不同部位的表达意义。方法将30只SD大鼠随机分为对照组(n=15)和模型组(n=15)。采用四氯化碳(CCL4)联合D-氨基半乳糖(D-GalN)和内毒素(LPS)注射法制备肝衰竭模型,使用B超监测筛选成功模型。检测血清ALT、AST、总胆红素(TBIL),采用HE染色观察肝组织病理学变化,采用免疫组化和免疫印迹法检测肝组织胞浆蛋白、胞核蛋白、总蛋白APE1/Ref1的表达。结果本组实验,经B超筛选模型成功率达80%;正常对照组血清ALT、AST和TBIL水平分别为(56.5±16.0)U/L、(178.7±36.5)U/L和(3.2±0.6)μmol/L,均显著低于模型组【(620.4±347.5)U/L、(1077.7±666.1)U/L和(21.2±16.9)μmol/L,均P<0.05】;免疫组化法检测模型组肝组织APE1/Ref1表达水平为(6.8±1.3)IOD,显著低于正常对照组的【(8.1±1.2)IOD,P<0.05】,模型组胞浆蛋白水平为(0.91±0.08)IOD,明显高于正常对照组的【(0.18±0.01)IOD,P<0.01】,而核蛋白和总蛋白水平分别为(0.71±0.01)IOD和(0.92±0.03)IOD,均明显低于正常对照组【(1.41±0.04)IOD和(1.15±0.01)IOD,P<0.05】。结论慢加急性肝衰竭大鼠肝内APE1/Ref1表达呈现由细胞核内转向胞浆内表达的特征,总体表达呈下降水平。Objective To investigate the expression of multifunctional protein apurinic/ apyrimidinic endonuclease 1（APE1/Ref1） in different parts of liver cells in rats with acute-on-chronic liver failure（ACLF）.Methods 30 SD rats were random1 y divided into normal control（n=15）,and ACLF model group（n=15）. ACLF models were established by intraperitoneal injections of CCL4 combined with D-Gal N and LPS and the establishment of liver failure model was confirmed by ultrasonography. Serum ALT,AST,and TBIL were detected by automatic biochemistry analyzer,pathologic changes in liver were revealed by HE staining,and APE1/Ref1 expression in cytoplasmprotein,nucleoprotein and total protein were detected by immunohistochemistry（IHC） and western blot. Results Success rate of model establishment screened by B ultrasound in this series was as high as80%;serum ALT,AST and TBIL in control group were significantly lower than those in the model group [（56.5±16.0） U/L vs.（620.4±347.5） U/L,（178.7±36.5） vs.（1077.7±666.1） U/L,（3.2±0.6） μmol/L vs.（21.2±16.9） μmol/L,respectively,P〈0.05 for all];the expression of APE1/Ref1 in model group was significantly lower than in the control[（6.8±1.3） IOD vs.（8.1±1.2） IOD,P〈0.05];APE1/Ref1 in cytoplasmprotein in model group was significantly higher than that in control group[（0.91±0.08） IOD vs.（0.18±0.01） IOD,P〈0.01],while APE1/Ref1 in nucleoprotein and total protein in model group were lower than those in control group [（0.71±0.01） IOD vs.（1.41±0.04） IOD,and（0.92±0.03） vs.（1.15±0.01） IOD,respectively,P〈0.05 for both]. Conclusion APE1/Ref1 are co-expressed in hepatocellular cytoplasm and nucleus,presenting transferred expression from intranucleus to intracytoplasm with a declined expression manner.

关 键 词：肝衰竭 脱嘌呤/脱嘧啶核酸内切酶 DNA损伤 氧化应激 大鼠 

分 类 号：R575.3[医药卫生—消化系统]