肝源性磷脂酰胆碱的分离纯化及其对肝癌细胞抑制作用的评价

Separation and Purification of Phosphatidylcholine in Swine Liver and Its Inhibition Effect on Proliferation of Rat Hepatoma Cells

采用氧化铝柱色谱法以95%乙醇作为洗脱剂分离纯化了肝源性磷脂酰胆碱(PC),并采用GF254硅胶板薄层色谱法以氯仿-甲醇-水(65∶25∶4,体积比)为展开剂测定了PC的纯度。结果表明以95%乙醇洗脱氧化铝色谱柱中的磷脂时,PC可完全与磷脂酰乙醇胺分离,其纯度和得率均高于80%。采用四甲基偶氮唑蓝比色法(MTT法)测定了不同浓度的PC对大鼠肝癌CBRH-7919细胞增殖的影响,并与人白血病K562细胞做比较,发现肝源性PC对肝癌细胞的生长具有明显的抑制作用,且存在时间-剂量依赖关系。

Phosphatidylcholine （PC） in crude phospholipids from swine liver was separated and purified by using Al2O3 column chromatography with 95% alcohol as eluent. The purity was determined by thin layer chromatography on GF254 silica gel plate and with chloroformmethanol-water （65：25：4,v/v） as developing agent. The results showed that PC was completely separated from phosphatidylethanolamine （PE） by the elution with 95% alcohol, and its purity and yield reached more than 90% and 80% with a elution volume of 225 mL, and 87.6% and 87.3% with a elution volume of 425 mL respectively. The effect of the PC with different concentrations on the proliferation of rat hepatoma cell line （CBRH-7919） was determined by microculture tetrazolium （MTT） assays in vitro and was compared with that of human leukemia cell line K562. Result shows that the PC derived from the liver inhibited the growth of CBRH- 7919 cells significantly. It suggested that PC derived from animal liver might function as a specific inhibitor for hepatoma cells in a concentration dependent manner.