人参总皂苷诱导HL-60细胞凋亡作用及对Caspase-3、Fas表达的影响

目的 探讨人参总皂苷诱导HL-60细胞凋亡的作用机制及其对凋亡相关基因Caspase-3、Fas表达的影响.方法 采用人参总皂苷0、100、200及400 μ g/ml作用于HL-60细胞,并在不同时间内用MTT法检测人参总皂苷对HL-60细胞的抑制率;48h后予普通光镜、荧光显微镜检测HL-60细胞形态的变化;予流式细胞仪、DNA凝胶电泳检测HL-60细胞凋亡;予RT-PCR法检测Caspase-3、Fas基因表达的变化.结果 人参总皂苷抑制HL-60细胞生长呈剂量及时间依赖性;在100～400 μ g/ml时,HL-60细胞凋亡逐渐增加,可见典型的凋亡细胞形态和明显的DNA梯度;在该浓度范围内,Caspase-3、Fas表达逐渐增加.结论 一定浓度的人参总皂苷可诱导HL-60细胞发生凋亡,且细胞凋亡率与人参总皂苷浓度呈一定的量效依赖关系,Caspase-3、Fas基因的表达变化在人参总皂苷诱导HL-60细胞凋亡中可能起重要作用.

Effect of total saponins of panaxginseng on Caspase-3 and Fas genes expression in HL-60 cell apoptosis induced

Objective To study the effect of the total saponins of panaxginseng （TSPG） on HL-60 ceils with expression change of apoptosis related genes caspase-3 and fas. Methods The suppressing rate was assayed by MTT; The morphology of HL-60 cells was observed under normal and fluorescence microscopes; the percentage of apoptotic HL-60 cells was essayed by flow cytometry （FCM） and the DNA ladder was observed by DNA agarose gel electrophoresis; the expression changes of caspase-3 and fas mRNAs were examined by RT-PCR, after HL-60 ceils were treated with TSPG at final concentrations of 0, 100,200,and 400 μ g/ml for 48 hours. Results TSPG inhibited the growth of HL-60 cells in dose-and time-dependent manner; The percentage of apoptotic HL-60 cells went up as the dose increased;The typical apoptotic cell morphology and the appearance of apoptotic DNA ladder were observed when treated with 100-400 μ g/ml TSPG for 48h.At the same time and same range of concentration, the expression of caspase-3 and fas mRNA increased gradually. Conclusion It suggested from the results that at certain range of concentration, apoptosis or necrosis could be induced by TSPG on HL-60 cells, and the percentage of apoptosis was dose-dependent, The effect on up-regulation of caspase-3 and fas mRNA probably plays important role in inducing apoptosis of HL-60 cells by TSPG.