钩藤中乌索酸和钩藤碱对肝癌细胞株HepG2的影响

Effects of Ursolic Acid and Rhynchophylline in Uncariae Ramulus Cum Uncis on Human Hepatoma HepG2 Cells

目的观察钩藤中乌索酸和钩藤碱对人肝癌细胞株HepG2的影响,探讨其抗肝癌的机制。方法将50、25、12.5、6.25μmol/L乌索酸、钩藤碱分别与人肝癌细胞株HepG2共同培养24、48、72 h后,以1%DMSO为阴性对照组,CCK-8法检测HepG2细胞增殖,吖啶橙荧光染色观察50μmol/L乌索酸、钩藤碱和1%DMSO作用48 h后HepG2细胞形态。结果乌索酸对HepG2细胞增殖有明显的抑制作用,并且其抑制率与作用时间、药物浓度呈正相关,钩藤碱对HepG2细胞增殖抑制作用较弱。病理观察结果显示,钩藤碱组大部分细胞出现细胞皱缩、包浆致密、核染色边集等凋亡早期改变,而乌索酸组大部分细胞出现核裂解、凋亡小体等凋亡晚期改变。结论钩藤中乌索酸和钩藤碱对人肝癌细胞株HepG2增殖有抑制作用,能诱导HepG2细胞凋亡。

Objective To investigate the effects of ursolic acid and rhynchophylline in Uncariae Ramulus Cum Uncis on human hepatoma HepG2cells;To discuss its antihepatoma mechanism.Methods Culture the human hepatoma HepG2cells with50,25,12.5,6.25μmol/L ursolic acid,rhynchophylline for24,48,72hours respectively.1%DMSO was set as negative control group.CCK-8based cytotoxicity assay was used to detect the growth of HepG2.Acridine orange fluorescent staining was used to observe human hepatoma HepG2cells cultured with50μmol/L ursolic acid,rhynchophylline and1%DMSO for48hours were observed.Results Ursolic acid significantly acted as a disincentive to the growth of HepG2cells,which was in positive correlation with time and concentration.Inhibited effect by rhynchophylline on HepG2cell proliferation was weaker than ursolic acid.Pathological observation showed that most of the cells in rhynchophylline group showed cell shrinkage,dense pulp coating and nuclear staining edge set,and most of the cells in the ursolic acid group showed apoptosis late changes,such as nuclear cleavage and apoptotic bodies.Conclusion Ursolic acid and rhynchophyllinein Uncariae Ramulus Cum Uncis can inhibit the proliferation of HepG2cells and can induce apoptosis.