论文部分内容阅读
【目的】探讨六味地黄丸主要入血成分处理的淋巴上清对大鼠肝癌细胞CBRH7919缝隙连接功能的影响,以期阐明其增强自杀基因疗法杀伤肝癌细胞效应的作用机制。【方法】取大鼠脾淋巴细胞,加入六味地黄丸的5种主要入血成分,体外培养后取含药淋巴上清,采用流式细胞术结合荧光示踪法分析其对细胞缝隙连接通讯功能(GJIC)的影响;利用Western blot法检测含药淋巴上清对大鼠肝癌CBRH7919细胞Cx43蛋白表达的影响;利用GJ阻滞剂甘草次酸(GA)观察阻滞GJIC后,含药淋巴细胞上清联合tk/GCV系统对细胞杀伤作用的影响。【结果】流式细胞术结合荧光示踪法结果表明,占培养液15%(体积分数)的含药淋巴上清能促进大鼠肝癌CBRH7919细胞的GJIC功能;Western blot法结果显示,含药淋巴细胞上清能提高大鼠肝癌CBRH7919细胞Cx43蛋白的表达水平;GA阻断GJIC功能后,含药淋巴细胞上清联合tk/GCV系统对细胞杀伤作用降低。【结论】六味地黄丸入血成分处理的淋巴上清可通过缝隙连接机制发挥对自杀基因杀伤大鼠肝癌细胞的增效作用。
【Objective】 To investigate the effect of lymphatic supernatant treated by Liuweidihuang Pills on the gap junction function of rat hepatocellular carcinoma cell line CBRH7919 in order to clarify its mechanism of enhancing the cytotoxic effect of suicide gene therapy on hepatocellular carcinoma cells. 【Method】 Five kinds of main blood components of Liuweidihuangwan were taken from splenic lymphocytes of rats. Lymphocyte supernatants were obtained after culture in vitro. Flow cytometry and fluorescent tracing method were used to analyze the cell gap junctional communication (GJIC). Western blot was used to detect the effect of drug-loaded lymphoid supernatant on the expression of Cx43 protein in rat hepatocellular carcinoma CBRH7919 cells. GJIC was used to observe the effect of GJIC on drug-containing lymphocytes Effect of combined tk / GCV system on cytotoxicity. 【Results】 Flow cytometry combined with fluorescent tracing method showed that the drug-containing lymphatic supernatant accounted for 15% (volume fraction) of culture medium, and promoted GJIC function in rat hepatoma CBRH7919 cells. Western blot showed that drug-containing lymphatic The cell supernatant increased the expression of Cx43 protein in rat hepatocellular carcinoma CBRH7919 cells. After GA blocked the function of GJIC, the cytotoxicity of drug-containing lymphocyte supernatant combined with tk / GCV system decreased. 【Conclusion】 The lymphatic supernatant of Liuweidihuangwan infusing ingredient can exert the synergistic effect on suicide gene-killing rat hepatoma cells through the gap junction mechanism.