益气活血软坚解毒方对荷H_(22)615小鼠肝癌细胞凋亡相关基因Fas、FasL、Bcl-2、Bax、P^(53)、NF-κB表达的影响  

Effect of Yiqihuoxue,Ruanjianjiedu recipe on expression of apoptosis regulate gene in mice with H_(22)615 tumor

作　　者：李东涛[1] 孙桂芝[2] 吴志奎[2] 李杰[2] 吕新霞[2] 陈玉英[2] 裴迎霞[2] 祁鑫[2] 

机构地区：[1]济南军区青岛第一疗养院,山东青岛266071 [2]中国中医科学院广安门医院

出　　处：《中西医结合肝病杂志》2010年第6期346-349,362,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金资助项目(No.30271656)

摘　　要：目的:研究益气活血软坚解毒(YHRJ)方对荷H22615小鼠肝癌细胞凋亡调控基因Fas、FasL、Bcl-2、Bax、P53、NF-κB表达的影响。方法:将动物分组为NS组(正常对照组)、DDP组(顺铂组)、YHRJ等效剂量组、YHRJ高剂量组、YHRJ等效剂量+DDP组、YHRJ高剂量+DDP组,应用免疫组化、原位杂交、RT-PCR等方法对用药后的荷H22615小鼠肝癌细胞凋亡主要相关基因Fas、FasL、Bcl-2、Bax、P53、NF-κB蛋白与mRNA表达进行检测。结果:免疫组化检测结果显示:与NS组比较,各加药组均能增高Fas、Bax基因蛋白表达(均P<0.01),均能降低NF-κB蛋白表达(P<0.05或P<0.01)及降低突变型P53基因蛋白表达(P<0.01)。与NS组比较,YHRJ等效剂量+DDP组能明显降低FasL基因蛋白表达(P<0.01)。与DDP组比较,中药组及中药含DDP组能明显降低FasL及突变型P53基因蛋白表达(均P<0.01)。与DDP组比较,YHRJ等效剂量组、YHRJ高剂量组、YHRJ高剂量+DDP组能降低NF-κB蛋白表达(均P<0.01)。与DDP组比较,YHRJ高剂量组及中药含DDP组明显增高Bax基因蛋白表达(均P<0.01)。与NS组与DDP组比较,中药组及中药含DDP组能明显降低Bcl-2基因蛋白表达(P<0.05或P<0.01)。原位杂交检测NF-κB mRNA表达结果:与NS组及DDP组比较,YHRJ等效剂量组及中药含DDP组明显降低NF-κB mRNA表达(均P<0.01)。RT-PCR检测Bax、Bcl-2 mRNA表达结果:与NS组及DDP组比较,YHRJ等效剂量组能明显增高Bax基因mRNA表达(均P<0.001)。与NS组比较,各加药组能明显降低Bcl-2 mRNA表达(P<0.05或P<0.001);与DDP组比较,YHRJ高剂量组能明显降低Bcl-2 mRNA表达(P=0.01)。结论:YHRJ方能够提高荷H22615小鼠肝癌细胞凋亡促进基因Fas、Bax表达,减低细胞凋亡抑制基因FasL、Bcl-2表达,降低细胞凋亡保护基因突变型P53蛋白与NF-κB基因表达,这些作用是诱导肝癌细胞凋亡的分子基础。Objective：To study the effect of Yiqi,Huoxue,Ruanjian Jiedu（YHRJ） Recipe on the expression of hepatocarcinoma cell,apoptosis regulate gene such as Fas,FasL,Bcl-2,Bax,P53,NF-κB in mice with H22615 Tumor.Methods：The mice were randomly divided into 6 groups：NS group,DDP group,YHRJ equivalent dosage group,YHRJ high dosage group,YHRJ equivalent dosage＋DDP group,YHRJ high dosage＋DDP group.Immunohistochemistry,situhybridization and RT-PCR were used to detect the key regulator genes of HCC apoptosis,Fas,FasL,Bcl-2,Bax,P53,NF-κB protein and mRNA in mice with H22615 tumor after 13 days treatment.Results：The immunohistochemistry results showed：compared with NS group,each adding medicine group could enhance the expression of Fas protein（P0.01） and YHRJ equivalent dosage＋DDP group could decrease obviously the expression of FasL protein（P0.01）.Compared with DDP group,Chinese medicines groups and Chinese medicines＋DDP groups could decrease obviously the expression of FasL protein（P0.01）.Compared with NS group,each adding medicine group could decrease obviously the expression of NF-κB protein（P0.05 or P0.01）;Compared with DDP group,YHRJ equivalent dosage group,YHRJ high dosage group,YHRJ high dosage＋DDP group could decrease obviously the expression of NF-κB protein（P0.01）.Compared with NS group,each adding medicine group could decrease obviously the expression of mutated P53 protein（P0.01）;Compared with DDP group,Chinese medicines groups and Chinese medicines＋DDP groups could decrease obviously the expression of mutated P53 protein（P0.01）.Compared with NS group,each adding medicine group could enhance obviously the expression of Bax protein（P0.01）;Compared with DDP group,YHRJ high dosage group and Chinese medicines＋DDP groups could enhance obviously the expression of Bax protein（P0.01）.The situhybridization detect the expression of results NF-κB mRNA,the results showed：compared with NS group and DDP group,YHRJ equivalent dosage group and Chinese medicines＋D

关 键 词：肝癌 益气活血软坚解毒方/药效学 细胞凋亡 基因调控 实验研究 

分 类 号：R735.7[医药卫生—肿瘤]