六味地黄丸调控肝癌CBRH7919细胞Cx43表达的体外研究

Regulatory Effect of Liuwei Dihuang Pills on Connexin 43 Expression in Hepatocarcinoma CBRH7919 Cells in Vitro

【目的】探讨六味地黄丸对大鼠肝癌CBRH7919细胞缝隙连接蛋白(Cx)43表达的影响。【方法】应用血清药理学方法制备六味地黄丸含药血清,分别采用蛋白免疫印迹(Western blot)法、异硫氰酸荧光素(FITC)标记间接免疫荧光染色—激光共聚焦显微镜技术、流式细胞术检测不同剂量(体积分数0%、2.5%、5%、10%)六味地黄丸含药血清组CBRH7919细胞Cx43蛋白的表达,采用实时定量逆转录—聚合酶链反应(RT-q PCR)检测各组肝癌细胞Cx43 m RNA的表达。【结果】与对照组(0%六味地黄丸含药血清)比较,体积分数为2.5%、5%、10%的六味地黄丸含药血清均能明显提高肝癌CBRH7919细胞Cx43蛋白及m RNA表达水平。【结论】六味地黄丸可能是通过促进肝癌CBRH7919细胞Cx43蛋白及m RNA的表达,改变Cx43膜定位,改善细胞间缝隙连接通讯功能,从而发挥抑制肝癌的作用。

Objective To investigate the effect of Liuwei Dihuang Pills（LDP） on the expression of connexin 43（Cx43） in rat hepatocarcinoma CBRH7919 cells. Methods LDP-containing serum was prepared by serum pharmacological method. The protein expression of Cx43 in CBRH7919 cells was determined by Western blotting method, indirect immuno-fluorescent staining labeled by fluorescein isothiocyanate（FITC）-laser scanning confocal microscope technology and flow cytometry. And the m RNA expression of Cx43 in the hepatoma cells was detected by reverse transcription real-time quantitative polymerase chain reaction（RT-q PCR）. Results Compared with the blank serum control group,the protein and m RNA expression levels of Cx43 were increased by volume fraction of 2.5%, 5%, 10% LDP-containing serum（P〈0.05 or P〈0.01）. Conclusion LDP play an anti-cancer role through promoting the expression of Cx43 in CBRH7919 cells, altering Cx43 membrane location,and improving the gap junctional intercellular communication（GJIC）.