健脾解毒方对二乙基亚硝胺诱导的肝癌大鼠肝组织CCR1和Cyclin D1 mRNA表达的影响  

Effects of Jiampi Jiedu recipe on expression of CCR1 and Cyclin D1 in rats with exprimental hepatocarcinogenesis induced by DEN

作　　者：张斌[1] 李琦[2] 殷佩浩[2] 赵成根[2] 高虹[2] 范忠泽[2] 

机构地区：[1]复旦大学附属中山医院青浦分院肝病科,上海201700 [2]上海中医药大学附属普陀医院肿瘤科

出　　处：《中西医结合肝病杂志》2013年第1期36-38,41,共4页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：上海市普陀区科委自主创新基金课题资助项目(No.200937);上海市嘉定区卫生局基金资助(NO.KYXM2010-12-03)

摘　　要：目的:探讨二乙基亚硝胺(diethylnitrosamine,DEN)诱导大鼠肝癌过程中肝组织CC趋化因子受体1(CCR1)、细胞周期蛋白D1(Cyclin D1)的表达规律及健脾解毒中药的干预机制。方法:雄性Wistar大鼠,随机分为正常组(n=25)、模型组(n=40)及中药组(n=40)。除正常组外,其他组大鼠在1～12周饮用含DEN 80mg/L的饮水以诱癌(8mg.kg-1.d-1),共12周。中药组大鼠同时给予含生药1.75g/ml的健脾解毒方灌胃(10ml/kg),正常组大鼠给予10ml/kg生理盐水灌胃,1次/d,共12周。于4、8、12、16周时相点,各组随机取5只大鼠剖腹取肝,20周时剩余大鼠全部剖腹取肝,观察肝脏外观,计算病死率和腹水生成率,肝组织进行HE染色,应用RT-PCR半定量检测肝组织CCR1、Cyclin D1 mRNA的表达。结果:在20周实验结束时,正常大鼠无死亡,而模型大鼠死亡率为42.5%(17/40),中药组死亡率为17.5%(7/40);16周～20周时模型组大鼠腹水发生率为87.5%(7/8),而中药组腹水发生率为44.4%(8/18),中药组与模型组差异有显著性意义(P<0.05)。肝组织HE染色表明,至第20周成瘤率达到100%。RT-PCR结果显示,与正常组比较,模型组各时段CCR1 mRNA均显著上调(P<0.01),与模型组比较,中药组除第8周外,其余各时段CCR1 mRNA均显著下调(P<0.01);与正常组比较,模型组大鼠除第4周及8周的Cyclin D1表达下调外,其余均出现显著上调,与模型组比较,中药组大鼠第4周、8周及12周的Cyclin D1表达上调,在16周时出现显著下调(P<0.01),第20周时两组差异不明显。结论:CCR1、Cyclin D1的表达上调与二乙基亚硝胺诱导肝癌形成密切相关,健脾解毒中药可以较好延缓肝癌的发生和发展,可能与下调CCR1关系密切,但下调Cylin D1的作用不明显,值得进一步深入研究。Objective： To investigate the mechanism of CCR1, Cyclin D1 expression and prevention effects of Jianpi Jiedu Recipe on experimental hepatocarcinoma rats induced by diethylnitrosamine （DEN） . Methods ： Male wistar rats were random- ly divided into normal control group （ n = 25 ）, model group （ n = 40 ） and prevention group （ n = 40 ） . In addition to the nor- mal group, other groups were given DEN drinking water （80ppm） at the dosage ofSmg ·kg^-1·d^-1 for continuous 12 weeks to induce hepatoma, while the prevention group were given Jianpi Jiedu Recipe （17.5g ~·kg^-1·d^-1, ig） , the normal group were given saline 10ml/kg, 1/d for 12 weeks. Each groups were randomly executed 5 rats at the 4th, 8th, 12th, 16th week respec- tively, and others rats were all killed in the 20th week. Hematoxylin-eosin staining （HE） were used to examine the changes of liver pathology, and the mRNA expression of CCR1 and CylinDl were tested by RT-PCR methods. Results： During 16 - 20 weeks, the mortality rate and ascite occur rate of normal control group was 0%, 0% respectively, while the model group was 42.5% , 87.5%, the prevention group was 17.5% , 44.4% , （P 〈0.05） . By the end of the 20th week , the incidence of HCC was 100% in model group and prevention group. To compared with normal group, the CCR1 mRNA expression of model group were significantly increased （P 〈 0. 01 ） in each week, while the CCR1 mRNA expression of prevention group were de- clined except the 8th week （P 〈0. 01, vs model group） . At the same time, the CylinD1 mRNA expression of model group were also significantly increased except the 4th week and 8th week （P 〈 0. 01, vs normal group） . To compared the model group, the CylinD1 mRNA expression of prevention group were increased at the 4th week, 8th week and 12th week, but CylinD1 expression of 16th week were declined （P 〈 0. 01 ） . AT the 20th week, there was no obvious difference between model group and prevention group. Conclusion ： The increased

关 键 词：原发性肝癌 二乙基亚硝胺 大鼠 健脾解毒方 药理作用 CC趋化因子受体1 细胞周期蛋白D1 

分 类 号：R285.5[医药卫生—中药学]