叶下珠复方Ⅱ号对二乙基亚硝胺诱导大鼠肝癌形成的抑制作用及机制  

Inhibitory Effect and Mechanism of Compound Phyllanthus Urinsria Ⅱ(CPU Ⅱ) on Formation of Liver Cancer in Rats Induced by DEN

作　　者：罗来育 陈斯泰[1] 李常青[1] 李小翚[2] 黄玉影 

机构地区：[1]广州中医药大学热带医学研究所,广州510405 [2]广州中医药大学中药学院,广州510405

出　　处：《中国实验方剂学杂志》2016年第8期126-132,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373524);广东省科技计划重点项目(2011A030100008)

摘　　要：目的:观察叶下珠复方Ⅱ号抑制二乙基亚硝胺(DEN)诱导的大鼠肝癌形成效果,并探讨其作用机制。方法:将240只SPF级雄性Wistar大鼠随机分为正常组,模型组,叶下珠复方Ⅱ号高、中、低剂量(23.14,11.57 g·kg-1)组,喃氟啶组,采用DEN诱导肝癌大鼠模型,造模18周,同时给药进行干预,每6周各组处死大鼠8只,观察大鼠一般情况,肝功能指标变化和肝癌结节形成情况,镜下观察大鼠肝组织病理变化,放免法检测大鼠血清白细胞介素-6(IL-6)含量;采用实时定量PCR(qRTPCR)法、免疫印迹法(Western blot)分别检测肝组织IL-6信号通路及microRNA let-7a调控网络基因的mRNA和蛋白表达改变。结果:实验第18周末,叶下珠复方Ⅱ号高、低剂量组大鼠肝癌结节数较模型组明显减少(P<0.05);叶下珠复方Ⅱ号高、低剂量组大鼠血清IL-6,谷丙转氨酶(ALT),谷草转氨酶(AST)水平与模型组比较均明显降低(P<0.05);与模型组比较,叶下珠复方Ⅱ号高剂量组大鼠肝组织microRNA let-7a表达量明显上升,转录因子蛋白家族(NF-κB-p65),IL-6,信号转导与转录激活因子3(STAT3),Harvery鼠肉瘤病毒Ras基因(Ras)和原癌基因(C-myc)mRNA表达量均明显降低(P<0.05)。实验第18周,叶下珠复方Ⅱ号高、低剂量组STAT3和p-STAT3蛋白表达均明显低于模型组(P<0.05)。结论:叶下珠复方Ⅱ号对DEN诱导的大鼠肝癌形成具有明显的抑制作用,作用机制与上调microRNA let-7a表达和下调NF-κB-p65的表达,从而抑制IL-6的表达和IL-6/STAT3信号通路活化有关。Objective： To observe the inhibitory effect of Compound phyllanthus Urinsria Ⅱ（ CPUⅡ）on formation of liver cancer in rats induced by diethylnitrosamine（ DEN）,and explore its anti-cancer mechanism.Method： Two hundred and forty SPF grade Wistar male rats were randomly divided into normal group,model group,CPU II high dose group（ 23. 14 g·kg- 1）,low dose group（ 11. 57 g·kg- 1）,and tegafur group. Rat models of liver cancer were induced by DEN for eighteen weeks,and medicine intervention was done at the same time according to the experimental design for eighteen weeks. 8 rats were sacrificed in each group after treatment for every 6 weeks,to observe the general condition,hepatic function index and nodule formation of liver cancer of the rats. Pathologic changes of liver tissues were observed under microscope. Radioimmunoassay method was used to determine the levels of interleukin（ IL）-6 in serum of rats. Gene expression levels of mRNA and protein related to IL-6 signal pathway and microRNA let-7a regulatory network of the liver tissues were detected respectively by real-time fluorescent quantitative PCR and Western blot method. Result： At the end of eighteenth week,the number of nodules of liver cancer in rats of CPU II high dose group and low-dose group was significantly reduced as compared with that in the model group（ P 〈 0. 05）; levels of IL-6,ALT and AST in serum of rats in CPU II high dose group and low dose group were significantly decreased（ P 〈 0. 05）; the expression level of microRNA let-7a in liver tissues increased obviously in CPU II high dose group as compared with that in model group,while the mRNA expression levels of NF-κB-p65,IL-6,STAT3,RAS and c-myc genes were significantly decreased（ P 〈0. 05）. The protein expression levels of STAT3 and p-STAT3 were also obviously reduced in CPU II high dose group and low dose group as compared with that in model group（ P 〈 0. 05）. Conclusion： CPUⅡcan effectively inhibit the formation of liver cancer in r

关 键 词：肝细胞癌 白细胞介素-6 炎症 微小RNA 

分 类 号：R285.5[医药卫生—中药学]