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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 extract total saponins from Clematis (TSRC) and to study the mechanism of TSRC on HL60 cells in vitro in acute promyelocytic leukemia HL60 cells. Methods: TSRC of 25-400 mg · L-1 was co-cultured with HL60 cells in vitro. The proliferation of HL60 cells was detected by WST-1 assay. Cell morphology and transmission electron microscope were used to observe the proliferation of TS60 cells. Annexin V / PI double staining The apoptosis of HL60 cells induced by TSRC was studied. The differentiation of HL60 cells induced by TSRC was studied by Wright’s-Giemsa staining, NBT reduction and CD11b immunocytochemistry. Results: 50 ~ 400 mg · L-1 TSRC significantly inhibited the proliferation of HL60 cells in vitro, showing a dose-time-dose-response relationship. The effects of TSRC at different concentrations for 24 h on the apoptosis of HL60 cells were statistically significant The significance of the induction of differentiation is not obvious. CONCLUSION: TSRC can effectively inhibit the proliferation of HL60 cells and induce the apoptosis of HL60 cells in vitro, but not induce differentiation of HL60 cells.