香附超临界CO_2提取物体外抗肝癌作用

Study on the anti hepatoma activity of cyperus rotundus by supercritical CO_2 fluid extraction in vitro

为研究香附超临界CO_2萃取物(XF)的体外抗肝癌活性,采用MTT比色法研究XF对人肝癌细胞HepG2、人正常肝细胞LO2的杀伤作用及其量-效和时-效关系;采用Annexin V-EGFP/PI双染、Rh123染色测定线粒体膜电位(Δψ)分析细胞凋亡及其机制.结果表明:XF对HepG2细胞具有强力杀伤作用,且呈明显的量-效和时-效关系.相比对肿瘤细胞和正常细胞均有较大毒性的阳性药顺铂(cis-Dichlorodiamineplatinum,DDP),XF对LO2细胞毒性较小,相对安全.Annexin V-EGFP/PI双染结果显示XF诱导细胞凋亡.Rh 123荧光染色检测表明XF引起Δψ崩溃.表明XF具有良好的体外抗肝肿瘤作用,其机制是破坏线粒体导致内源性凋亡,值得开发成安全有效的抗肝肿瘤药.

This study was performed to investigate anti hepatoma activity of the effective component of rhizoma cyperi with the technique of supercritical CO2 fluid extraction （XF） in vitro. The cytotoxicity of XF both on human hepatoma cells HepG2 and on normal human hepatocytes LO2 were evaluated by MTT assay. Annexin V-EGFP/PI double staining and Rhodamine123 （Rh123） staining were tested in the preliminary study of its mechanism. Results. XF showed a significant dose dependent and time dependent cytotoxicity to the tumor cells. Compared to the positive control drug cisplatin （DDP）, which has a great toxicity on both tumor and normal cells, XF is less toxic to LO2 cells. Annexin V-EGFP/PI double staining showed that XF could strongly induce apoptosis. Rh123 method indicated that XF caused the loss of mitochondrial membrane potential. In Conclusion. XF has a remarkable anti hepatoma effect in vitro and is expected to be developed into an effective anti hepatoma drug.