六味地黄汤及拆方对小鼠黑色素瘤B16细胞胸苷激酶/更昔洛韦自杀基因疗法旁观者效应的影响  

Effect of Liuwei Dihuang Decoction and its decomposed prescription on the bystander effect of HSV-tk/GCV suicide gene therapy in mouse melanoma B16 cells

作　　者：赵婷秀[1] 潘啟暘 孙渊 王幸鹏 刘坤东 吴绍锋 杜标炎[1] 吴彤[2] ZHAO Tingxiu;PAN Qiyang;SUN Yuan;WANG Xingpeng;LIU Kundong;WU Shaofeng;DU Biaoyan;WU Tong(School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学基础医学院,510006 [2]广州中医药大学第一临床医学院,510006

出　　处：《环球中医药》2025年第3期405-412,共8页Global Traditional Chinese Medicine

基　　金：广东省中医药局科研项目面上项目(20221119);国家自然科学基金面上项目(81873146)。

摘　　要：目的观察六味地黄汤及拆方对小鼠黑色素瘤B16细胞胸苷激酶(herpes simplex virus-thymidine kinase,HSV-tk)/更昔洛韦(ganciclovir,GCV)自杀基因疗法旁观者效应的作用机制及配伍规律。方法实验动物分对照组、GCV组、GCV+六味组、GCV+三补组、GCV+三泻组。采用小鼠双侧腋下分别移植B16-tk细胞和B16细胞、腹腔注射GCV的方法,建立小鼠黑色素瘤B16细胞HSV-tk/GCV自杀基因移植瘤模型。对照组腹腔注射生理盐水。各组给予相应药物灌胃给药14天,观察六味地黄汤及拆方对B16肿瘤的旁观者效应。ELISA法检测B16肿瘤和脾脏的IL-2浓度;观察B16肿瘤和脾脏的淋巴细胞分布情况。结果与对照组比较,各组B16-tk肿瘤生长均显著抑制。与GCV组比较,GCV+六味组和GCV+三补组B16肿瘤的生长明显抑制、肿瘤系数显著减少(P<0.01),IL-2浓度明显升高(P<0.01)、淋巴细胞浸润明显,其中,GCV+六味组CD_(8)^(+)T细胞浸润明显,GCV+三补组CD_(4)^(+)T细胞浸润明显;脾脏的IL-2浓度明显降低(P<0.01)、CD_(8)^(+)T细胞增多。而GCV+三泻组的B16肿瘤生长加速,肿瘤系数增加,肿瘤IL-2明显降低(P<0.01),肿瘤组织充血和出血明显,未见淋巴细胞浸润;脾脏IL-2明显升高(P<0.01)、未见CD_(8)^(+)T细胞。结论六味地黄汤可以明显增效小鼠黑色素瘤B16细胞HSV-tk/GCV自杀基因疗法旁观者效应,其机制可能与改善肿瘤和脾脏的IL-2浓度以及淋巴细胞数量有关,增效的主要物质在三补成分,而三泻成分拮抗旁观者效应。Objective To observe the mechanism and compatibility law of Liuwei Dihuang Decoction and its decomposed prescription on the bystander effect of HSV-tk/GCV suicide gene therapy in mouse melanoma B16 cells.Methods The experimental animals were divided into control group,GCV group,GCV+Liuwei group,GCV+San-bu(three tonics)group and GCV+San-xie(three eliminators)group.The HSV-tk/GCV suicide gene transplanted tumor model of mouse melanoma B16 cells was established by bilateral axillary respectively transplantation of B16-tk cells and B16 cells and GCV was injected intrabitoneally.Normal saline was injected intrabitoneally in the control group.Each group was given the corresponding drug intragastric administration for 14 days to observe the bystander effect of Liuwei Dihuang Decoction and its decomposed prescription on B16 tumor.IL-2 concentration of B16 tumor and spleen was detected by ELISA.The lymphocyte distribution of B16 tumor and spleen was observed.Results Compared with the control group,B16-tk tumor growth was significantly inhibited in all groups.Compared with the GCV group,The growth of B16 tumor was significantly inhibited,the tumor coefficient was significantly decreased(P<0.01),the IL-2 concentration was significantly increased(P<0.01),and the lymphocyte infiltration was obvious in the GCV+Liuwei group and the GCV+San-bu group,among which CD_(8)^(+)T cell infiltration was obvious in the GCV+Liuwei group and CD_(4)^(+)T cell infiltration was obvious in the GCV+San-bu group.The IL-2 concentration in the spleen was decreased significantly(P<0.01),and CD_(8)^(+)T cells was increased.In the GCV+San-xie group,The growth of B16 tumor was accelerated,tumor coefficient was increased,IL-2 was decreased significantly(P<0.01),The tumor tissue showed obvious congestion and bleeding,and no lymphocyte infiltration was observed.IL-2 was significantly increased in spleen(P<0.01)and CD_(8)^(+)T cells were not found.Conclusion Liuwei Dihuang Decoction can significantly enhance the bystander effect of HSV-tk/GCV suicide gene

关 键 词：六味地黄汤 拆方 黑色素瘤 自杀基因疗法 旁观者效应 

分 类 号：R285.5[医药卫生—中药学]