枸杞叶中总黄酮对人肝癌细胞HepG2增殖与凋亡的影响

研究枸杞叶中总黄酮对体外培养的人肝癌细胞HepG2增殖抑制及其细胞凋亡的影响.用不同质量浓度的枸杞叶总黄酮作用于体外培养的人肝癌细胞HepG2,用四甲基偶氮噻唑蓝(MTT)法检测HepG2细胞存活率,通过Hoechst33258染色法、AV/PI双染流式细胞术检测HepG2细胞凋亡情况,利用免疫细胞化学法、Western blot方法分析枸杞叶中总黄酮对HepG2细胞凋亡相关蛋白Bax、Caspase-3表达的影响.结果表明,枸杞叶中总黄酮分别作用于人肝癌细胞HepG2 48,72h均可明显抑制其增殖,并呈现显著的时间、剂量依赖性.高质量浓度枸杞叶总黄酮作用人肝癌细胞HepG2 48h后,HepG2细胞呈现出典型的细胞凋亡特征,凋亡率达67.3%.枸杞叶总黄酮作用于HepG2细胞一定时间后,可引起促凋亡蛋白Bax、Caspase-3的表达上调.枸杞叶中总黄酮可明显抑制人肝癌细胞HepG2增殖和诱导细胞凋亡,其作用机制可能是通过有效上调促凋亡蛋白Bax,破坏细胞内稳态环境,继而激活凋亡蛋白Caspase-3,最终引发人肝癌细胞HepG2凋亡.

Effects of Total Flavonoids in Lycium chinensis Leaves on Proliferation and Apoptosis of Hepatoma Cell line HepG2 in vitro

This article investigates the effects of total flavonoids in Lycium chinensis leaves on cell proliferation and apoptosis of human hepatocellular carcinoma （HepG2 cells）cultured in vitro.The total flavonoids in Lycium chinensis leaves ’ effect on HepG2 apoptosis related-protein Bax and Caspase-3 expression is analyzed in immunocytochemical method and Western blot with different concentration mass of doses of total flavonoids in Lycium chinensis leaves on hepatoma cell line HepG2 in vitro,with the survive rated of HepG2 detected with methyl thiazolyl tetrazolium （MTT）method,and the HepG2 Cell apoptosis detected in Hoechst33258 staining method and AV/PI Double color flow cytometry.The result shows that when Total Flavonoids in Lyciumchinensis Leaves affect Hepatoma Cell line HepG2 in both 48 and 72 hours can hold up the proliferation and it remarkably relies on time and doses.After 48 hours of Leaves affect Hepatoma Cell line HepG2’s effect on Hepatoma Cell line HepG2,the cell HepG2 appears typical cell apoptosis,the apoptosis rate reaches 67.3%.After a certain time of Total Flavonoids in Lycium chinensis Leaves affect HepG2 ’s,the pro-apoptotic protein Bax and Caspase-3 expression will increase.Total Flavonoids in Lycium chinensis Leaves can hold clearly the Proliferation of Hepatoma Cell line HepG2 in vitro and cause apoptosis.The mechanism of action is probably effectively up-regulating pro-apoptotic protein Bax,destroying the cells homeostasis environment,then activating apoptin Caspase-3 and at last causing the apoptosis of Hepatoma Cell line HepG2.