中文摘要：

目的：探讨六味地黄丸对大鼠肝癌CBRH7919细胞Cx26表达的影响，初步阐明其对自杀基因治疗的增效作用是否与缝隙连接机制相关。方法：应用血清药理学研究方法制备六味地黄丸含药血清。采用MTT法检测六味地黄丸含药血清联合单纯疱疹病毒-胸苷激酶／丙氧鸟苷（HSV-tk／GCV）自杀基因系统治疗的杀伤效应：应用蛋白印迹（Western blot）技术、间接免疫荧光法及荧光定量逆转录-聚合酶链反应（RT—PCR）技术检测其对CBRH7919细胞株Cx26蛋白及mRNA表达的影响。结果：①六味地黄丸含药血清联合自杀基因治疗组肝癌细胞存活率明显低于单纯自杀基因组（10％tk^＋／GCV加对照血清）（P〈0．01），且联合作用均具有协同性（Q〉1．15）；②Western blot检测显示六味地黄丸含药血清能提高Cx26蛋白的表达，并有明显的量效关系；③间接免疫荧光法（FITC）法共聚焦显微镜观测显示近似的结果：蛋白定位于细胞膜和胞浆，尤其在细胞膜的表达明显增多，含药血清组的Cx26蛋白荧光强度比对照组明显增加（P〈0．05）。结论：六味地黄丸增强自杀基因旁杀伤效应的机制与其促进肝癌细胞Cx26 mRNA及蛋白表达，增加膜上定位表达进而改善细胞间连接通讯CGJIC）功能有关。

英文摘要：

Objective： To investigate the regulation effect of medicated serum of Liuwei Dihuang Bolus on the expression of Connexin26 in rat hepatocarcinoma CBRH7919 cell, and clarity whether the mechanism of Liuwei Dihuang Bolus promoting the suicide gene therapy was related with the gap junction. Methods： Blood-serum pharmacology was used to prepare the medicated serum of Liuwei Dihuang Bolus. The lethal effect of the medicated serum of Liuwei Dihuang Bolus associated with the HSV - tk / GCV suicide gene therap system was detected by MTT assay. The expression of Cx26 and mRNA on rats hepatoma cell line CBRH7919 were detected by western blot, indirect immunofluorescence assay （ IIA ） and real-time fluorescence quantitative RT-PCR assay. Results： （1）The survival rate of hepatoma carcinoma cell in medicated serum combining with tk^＋/GCV group was significantly lower than that in suicide gene therapy group（10%tk^＋/GCV with control serum）（P 〈 0.01）, the lethal effect had obvious eooperativity （ Q〉1.15 ） ; （2）The result of western blot indicated that the medicated serum of Liuwei Dihuang Bolus can elevate the protein expression of Cx26, also the up-regulated effects showed obvious dose-effect relationship. （3）The IIA assay showed similar results： Cx26 protein was located on the cell membrane and cytoplasm, especially more located on cell membrane, and the fluorescence intensity of medicated serum group is much stronger than that of control-serum group （P 〈 0.05）. Conclusion： The mechanism of Liuwei Dihuang Bolus enhancing suicide gene bystander effect relates to that it can promote the expression Cx26 mRNA and protein on CBRH7919, make more Cx26 located on cell membrane, improve the gap junctional intercellular communication （GJIC）.