六味地黄丸入血成分对自杀基因杀伤肝癌细胞的增效作用  

Enhancing Effect of Constituents Absorbed into Blood after Oral Administration of Liuwei Dihuang Wan on Killing Hepatocarcinoma Cells with HSV-tk/GCV System

作　　者：杜标炎[1] 周瑶[2] 易华[1] 张广献[3] 谭宇蕙[3] 吴映雅[3] 郭玉荣[4] 罗惠[1] 

机构地区：[1]广州中医药大学病理教研室,广州510006 [2]湖南中医药高等专科学校形态教研室,湖南株洲412012 [3]广州中医药大学生化教研室,广州510006 [4]广西柳州市中医院,广西柳州545001

出　　处：《中国实验方剂学杂志》2013年第16期220-224,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072906)

摘　　要：目的:探讨六味地黄丸主要入血成分对单纯疱疹病毒胸苷激酶/丙氧鸟苷(HSV-tk/GCV)自杀基因系统杀伤大鼠肝癌细胞(CBRH7919)的增效作用。方法:按CBRH7919/tk占总细胞数10%的比例混合CBRH7919与CBRH7919/tk细胞,分别接种96孔板,每孔加细胞悬液100μL(2×103个/孔),设置对照组、低、中、高3种浓度的六味地黄丸入血成分组、GCV组、各浓度六味地黄丸入血成分联合GCV组;培养24 h后分别加入入血成分马钱素和丹皮酚(质量浓度均为25,50,100μmol.L-1),莫诺苷、5-羟甲基-2-糠酸、獐牙菜苷(浓度均为37.5,75,150μmol.L-1)5种化合物和5种化合物混合组分(质量浓度为0.006 7,0.013 5,0.027 g.L-1);及其混合组分(质量浓度为0.034 g.L-1)作用后淋巴细胞培养上清(占培养液体积分数7.5%,15%,30%),设6个复孔,37℃,5%CO2培养箱培养72 h后MTT法测定细胞存活率,检测六味地黄丸主要入血成分及淋巴细胞培养上清对自杀基因系统杀伤肝癌细胞的影响。结果:自杀基因系统联合马钱素、莫诺苷、5-羟甲基-2-糠酸、獐牙菜苷和丹皮酚5种化合物及5种化合物的混合组分,均未显示其对肿瘤自杀基因疗法的直接增效作用;但含药上清联合GCV组对肝癌细胞生长抑制率分别为(22.72±3.12)%,(26.68±1.92)%,(30.50±3.53)%;与单独自杀基因(10%tk/GCV)组抑制率(15.37±3.96)%比较,差异均有统计学显著性意义;q值分别为1.50,1.75,2.24,均大于1.15,具有协同增效作用,并呈一定的量效关系。结论:六味地黄丸主要入血成分能刺激淋巴细胞产生某些活性物质,并发挥了对HSV-tk/GCV自杀基因治疗系统对肝癌CBRH7919细胞杀伤的增效作用,这些淋巴细胞产生的内源性活性物质可能才是六味地黄丸对肿瘤自杀基因疗法增效作用的药效物质基础。Objective： To explore the enhancing effect on killing hepatocareinoma cells of CBRH7919 by herpes simplex virus thymidine kinase and ganciclovir （HSV-tk/GCV） system together with constituents absorbed into blood after oral administration of Liuwei Dihuang Wan. Method： MTT assay was used to detect growth inhibition of CBRH7919 after being treated with tk/GCV system and constituents including loganin, morroniside,methoxymethyl） -2-furoic acid, sweroside, paeonol and mixed drugs of these five compounds; also to detect growth inhibition of CBRH7919 after being treated with tk/GCV system and lymphocytic supernatant fluid activated by constituents. Result： There was no synergy killing effect on CBRH7919 between tk/GCV system and constituents; while there was synergy killing effect on CBRH7919 treated with tk/GCV system lymphocytic supernatant fluid activated by constituents together, witch showed significantly dose-effect relationship. Conclusion ： The active substrate activated by constituents of Liuwei Dihuang Wan has synergy effect on HSV-tk/ GCV system, and the endogenous active substrate may be the material basis of synergy effect of Liuwei Dihuang Wan on HSV-tk/GCV system.

关 键 词：肝癌 基因治疗 自杀基因 单纯疱疹病毒胸苷激酶 丙氧鸟苷系统 六味地黄丸 

分 类 号：R285.5[医药卫生—中药学]