七叶皂苷诱导HL-60细胞凋亡的研究

Apoptosis of HL-60 Cells Induced by Aescinate

本研究观察七叶皂苷对髓系白血病HL-60细胞的抑制和诱导凋亡作用。取对数期生长的HL-60细胞经饥饿处理后,加入不同浓度的七叶皂苷观察七叶皂苷抑制细胞生长的指数、形态学变化,用DNA片段凝胶电泳(DNA Ladder)和Annexin V/PI-FITC双标法分析诱导细胞凋亡的作用。结果表明:七叶皂苷抑制HL-60细胞生长,15-120mg/L的七叶皂苷处理细胞48小时后,存活率为(92.2±0.69)%-(8.2±0.96)%,均明显低于不加药的对照组(99.4±0.31)%(p均<0.05);七叶皂苷15-60mg/L处理细胞24小时后细胞发生凋亡。Annexin V/PI分析显示,给药组AnnexinV阳性细胞为(12.7±0.58)%-(65.4±1.30)%,明显高于对照组的(0.57±0.03)%(p均<0.01)。七叶皂苷处理的细胞DNA琼脂糖凝胶电泳显示典形的凋亡DNA梯形条带。结论:七叶皂苷能特异性地诱导HL-60细胞凋亡,此研究结果为七叶皂苷今后作为治疗白血病的辅助药物提供实验依据。

The aim of this study was to investigate the effects of aescinate on inhibition and apoptosis of HL-60 cell line from promyelocytic leukemia. HL-60 cells at logarithm phase were treated with aescinate. Cell survival rate and cell morphology were observed, and the cell apoptosis was analyzed by Annexin V/PI-FITC double labeling and DNA electrophoresis. The results showed that HL-60 cells could be inhibited in the presence of 15 - 120 mg/L of aescinate for 48 hours, survival rates were （92.2 ± 0.69 ） % - （8.2 ± 0.96） %, which were significantly lower than that of non- aescinate control （99.4 ±0.31 ）% （all p 〈0.01 ）. The apoptosis of cells could be induced by aescinate treatment at dosage of 15 -60 mg/L for 24 hours, the Annexin V positive cells accounted for （12.7 ±0.58）% - （65.4 ± 1.30）% which were significantly higher than that of non- aescinate control （0.57 ± 0.03） % （ all p 〈0.01 ）. The typical DNA ladder of HL-60 cells treated with aescinate was shown on the DNA electrophoresis pattern. It is concluded that aescinate can specifically induce apoptosis of leukemic HL-60 cells, which provides an experimental evidence for treatment of leukemia with aescinate as a supplementary agent to chemotherapy.