铁包金通过APE1调节凋亡相关蛋白防治慢加急性肝衰竭大鼠的作用  

Preventive and Therapeutic Effects of Radix Berchemiae Lineatae on Acute-on-chronic Liver Failure Rats Through Regulating APE1 Apoptosis-associated Proteins

作　　者：刁建新[1] 马文校[1] 戴凤翔 李铭链 张婷婷[1] 陆梓雯 杨运高[1] 

机构地区：[1]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2016年第6期794-799,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金(81173262);广东省大学生创新创业训练项目(201512121057)

摘　　要：目的观察铁包金对慢加急性肝衰竭的作用及其对无嘌呤/无嘧啶核酸内切酶1(APE1)、caspase3、bax、bcl-2的影响。方法取SD大鼠40只,随机分为4组,每组10只,分别为正常对照组、模型组、铁包金组(8 g·kg^(-1)·d^(-1))和安宫牛黄丸组(0.27 g·kg^(-1)·d^(-1))。采用四氯化碳(CCL4)联合D-氨基半乳糖(D-Gal N)和脂多糖(LPS)复制慢加急性肝衰竭模型。观察大鼠48 h内的病死率。全自动生化仪检测丙氨酸氨基转移酶(ALT)、天冬氨酸转移酶(AST)、血清总胆红素(TBIL)的水平。HE染色观察肝脏病理变化。Tunel染色观察肝细胞凋亡指数(AI)。免疫印迹检测APE1、caspase3、bax和bcl-2蛋白的表达。共聚焦显微镜观察APE1的原位表达。结果正常对照组、模型组、铁包金组和安宫牛黄丸组48 h病死率分别为0%、50%、20%和20%。与模型组比较,铁包金组显著降低ALT、AST、TBIL、AI、caspase3和bax的水平,差异有统计学意义(P<0.05);铁包金组可显著提高APE1、bcl-2的水平,差异有统计学意义(P<0.05);铁包金组作用效果与安宫牛黄丸组相当,差异无统计学意义(P>0.05)。结论铁包金对慢加急性肝衰竭具有一定的防治作用,可能是铁包金通过APE1/Ref^(-1)调控凋亡相关蛋白防治慢加急性肝衰竭。Objective To observe the therapeutic effects of Radix Berchemiae Lineatae（Tiebaojin）on acute-on-chronic liver failure（ACLF）rats,and to investigate its influence on regulating apurinic/apyrimidinic endonuclease 1（APE1）,caspase 3,bax,and bcl-2 protein.Methods SD rats were randomly divided into normal control group,model group,Tiebaojin group（8 g·kg^-1·d^-1）,and Angong Niuhuang Wan group（0.27 g·kg^-1·d^-1）.We adopted carbon tetrachloride（CCL4） combined with D-galactosamine（D-Gal N） and lipopolysaccharide（LPS）to establish ACLF rat model,and observed the rat mortality within 48 h.Automatic biochemical analyzer was used to detect the serum levels of alanine aminotransferase（ALT）,aspartate transaminase（AST）,and total bilirubin（TBIL）.HE staining was used to observe the liver pathology.Tunel staining was applied to observe the liver cell apoptotic index（AI）.APE1,caspase3,bax and bcl-2protein expression was examined by Western blotting,and in-situ expression of APE1 was observed by confocal microscopy.Results The 48-h mortality of normal control group,model group,Tiebaojin group,and Angong Niuhuang Wan group was 0 %,50 %,20 % and 20 % respectively.Compared with the model group,Tiebaojin group had significant effect on reducing ALT,AST,TBIL,AI,caspase3 and bax levels（P〈0.05）,increasing the levels of APE1 and bcl-2（P〈0.05）.The effects of Tiebaojin were similar to those of Angong Niuhuang Wan（P〈0.05）.Conclusion Tiebaojin exerts certain preventive and therapeutic effect on ACLF,and the effect is likely to be achieved through regulating APE1/Ref 1 apoptosis-associated protein.

关 键 词：铁包金 慢加急性肝衰竭 无嘌呤/无嘧啶核酸内切酶1 半胱天冬酶3 BAX bcl-2 

分 类 号：R285.5[医药卫生—中药学]