癌痛消方对DEN诱癌大鼠不同时间点凋亡相关蛋白的影响  

Study on the related apoptotic proteins at different time points in DEN induced hepatocarcinoma rats treated with Aitongxiao prescription

作　　者：胥冰[1] 黄峰[1,2] 韦艾凌 张红[1] 王小平[1] 田亚宁[2] 

机构地区：[1]陕西中医药大学,陕西咸阳712046 [2]陕西中医药大学附属医院,陕西咸阳712000 [3]广西中医药大学,广西南宁530000

出　　处：《华西药学杂志》2017年第5期489-492,共4页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:30960475;81172135);陕西省自然科学基础研究项目(No.2016JM8023;2016JM8150;2014JM4175);陕西省教育厅自然科学基础研究项目(No.07JK233;14JS025;11JK0686);陕西省中医药管理局科研项目(131DC016)

摘　　要：目的通过分析癌痛消方(AP)干预二乙基亚硝胺(DEN)诱癌大鼠模型在不同时间点的肝脏组织病理变化、细胞凋亡的相关蛋白表达变化,来探讨癌痛消方治疗原发性肝癌的机制。方法将50只♂Wistar大鼠随机分为预防组、模型组(每组20只)和空白对照组(10只),预防组和模型组的大鼠以64 ppm DEN饮水饲养,连续16周。同时,自诱癌之日起,预防组大鼠予以AP灌胃0.2 g·(100 g)^(-1),qd,每周连续用药6 d,给药周期为18周;模型与空白对照组正常饮食,饮水(自来水)。分别在4、8、12、16、18周时,随机各取预防组和模型组的4只大鼠、2只空白对照组的,断颈处死,取肝组织(阳性部位)免疫组化法检测Bax、Bcl-2、survivin、Fas、Fasl、caspase-3蛋白的表达情况。结果空白对照组未见survivin、casepase-3、Fas/Fasl、Bax、Bcl-2蛋白的表达;模型组各时间点均可见凋亡相关蛋白的表达,以凋亡抑制蛋白Bcl-2、survivin表达量较多;AP干预过程可以全程下调survivin、Bcl-2蛋白的表达,上调Fas、casepase、Bax蛋白的表达。结论 AP可以促进受损肝细胞或癌细胞凋亡,其预防和治疗肝癌机理是上调促凋亡蛋白、下调抑凋亡蛋白的表达,促进炎症细胞或肿瘤细胞凋亡。OBJECTIVE To investigate the anti-hepatocarcinoma mechanism of Chinese herbs Aitongxiao prescription（ AP） by analyzing the related apoptotic proteins and liver pathological changes at different time points in DEN induced hepatocarcinoma rats.METHODS Fifty male Wistar rats were divided into preventive and model groups,each of which contained 20 rats. 10 rats were set as control. Preventive and model groups were fed with 64 ppm DEN buffer for 16 weeks. Meanwhile,the preventive group was given AP 0. 2g·（ 100 g）^（-1）everyday for 18 weeks by gavage. Model and control groups were given regular diet without drug administration. At 4,8,12,16 weeks respectively,4 rats in preventive and model groups and 2 rats in control group were selected and sacrificed randomly. The liver tissues were collected and the expression of Bax,Bcl-2,survivin,Fas,Fasl and caspase-3 were detected by immunohistochemistry. RESULTS No immunostaining were shown for survivin,casepase-3,Fas/Fasl,Bax and Bcl-2 in control group. Model group expressed apoptosis related proteins at different time points,especially for anti-apoptotic proteins,Bcl-2 and survivin,while survivin and Bcl-2 were down-regulated and Fas,casepase,Bax were up-regulated in the AP administration group. CONCLUSION AP can promote the apoptosis of injured liver cells or cancer cells by up-regulating the expression of pro-apoptotic proteins and down-regulating the expression of anti-apoptotic proteins.

关 键 词：诱导性肝癌 大鼠模型 癌痛消 Bcl-2 Bax Survivin Casepase 二乙基亚硝胺 

分 类 号：R96[医药卫生—药理学]