威灵仙总皂苷对HL60细胞株体外作用的研究

Effect of Total Saponin from Radix Clematidis on Acute Promyelocytic Leukemia HL60 Cell Line in vitro

目的:提取威灵仙总皂苷(TSRC),体外作用于急性早幼粒细胞白血病HL60细胞,研究TSRC对HL60细胞的作用机制。方法:将25～400 mg·L-1的TSRC在体外与HL60细胞共同培养,WST-1法检测TSRC对HL60细胞的增殖抑制作用;采用细胞形态学、透射电镜观察细胞、AnnexinV/PI双染色研究TSRC诱导HL60细胞凋亡作用;采用Wright’s-Giemsa染色细胞形态学观察、硝基蓝四氮唑(NBT)还原试验、细胞免疫表型CD11b检测等方法研究TSRC诱导HL60细胞分化作用。结果:50～400 mg·L-1TSRC在体外对HL60细胞增殖有明显抑制作用,呈现剂量和时间的量效关系;不同浓度TSRC作用24 h后均能显著促进HL60细胞凋亡;但具有统计学意义的诱导分化现象不明显。结论:TSRC在体外可有效抑制HL60细胞增殖,并诱导HL60细胞凋亡,但对HL60细胞无明显的诱导分化作用。

Objective： To study the effect of total Saponin of Radix Clematidis （TSRC） on HL60 leukemia cell in vitro. Method： In vitro, HL60 cells were incubated with 25-400 mg ·L^-1TSRC, the proliferation inhibition rate of HL60 was tested by WST-1. Cell morphology, transmission electron microscope （TEM） and Annexin-V/PI were used to investigate the apoptosis of HL60 ceils. The differentiations of HL60 was analyzed by morphology of Giemsa stain, nitroblue tetrazdium （NBT） reduced method. The expressions of CD1 l b and CD33 induced by these drugs were measured by flow cytometry （FCM） at different time. Result： As compared with blank control group, Different concentrations from 50-400 mg·L^-1 TSRC could obviously inhibit the growth of HL60 ceils with time and dose dependence ; While no statistical significance in the differentiation phenomenon was observed. Conclusion： In vitro, HL60 cell＇ s proliferation inhibition and apoptosis could obviously induced by TSRC. But HL60 cells could not be induced differentiation by TSRC.