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目的探讨人参总皂甙(total saponins of panax ginseng,TSPG)对人急性髓系白血病细胞株KG1α增殖的影响。方法取对数生长期的KG1α细胞,调整密度为5×108/L,空白对照组予以常规培养;人参总皂甙TSPG组分别加入0.1、0.2、0.4、0.6、0.8 g/L TSPG。采用MTT比色法测定细胞存活率,瑞氏染色法以及光电镜观察细胞形态学改变。流式细胞仪测定细胞周期分布的变化以及细胞的凋亡率。免疫细胞化学的方法检测细胞中β-catenin、NF-κB p65蛋白表达的变化。RT-PCR半定量检测细胞中β-catenin mRNA和NF-κB p65 mRNA水平的变化。结果人参总皂甙TSPG在体外对KG1α增殖具有明显抑制作用。在0.1~0.8 g/L TSPG浓度范围内,其抑制作用呈浓度依赖性,在浓度0.4 g/L处达到细胞的半数抑制率,且在48 h抑制率达到高峰。0.4 g/L TSPG作用48 h后,可见细胞分散生长,数量明显减少;细胞明显分化或凋亡,产生核分裂象,胞质中产生凋亡小体以及空泡;与对照组相比,加药组KG1α细胞周期分布无明显变化(P>0.05);与对照组相比,加药组细胞凋亡率显著增高,其改变具有统计学意义(P<0.05);与对照组相比,加药组细胞内β-catenin蛋白因子表达降低其表达区域由胞核向胞膜转移,NF-κB p65因子的表达也明显减弱其表达区域由胞核向胞膜转移;与对照组相比,加药组β-catenin和NF-κB p65 mRNA的水平降低,其改变具有统计学意义(P<0.05)。结论 TSPG可抑制KG1α细胞增殖诱导细胞凋亡,且TSPG抑制KG1α细胞增殖的作用机制可能与其抑制β-catenin及NF-κB p65蛋白表达,降低β-catenin和NF-κB p65 mRNA的水平有关。
Objective To investigate the effects of total saponins of panax ginseng (TSPG) on the proliferation of human acute myeloid leukemia cell line KG1α. Methods KG1α cells in logarithmic growth phase were weighed and adjusted to a density of 5 × 108 / L. The blank control group was routinely cultured. Total ginsenoside TSPG group was added with 0.1, 0.2, 0.4, 0.6 and 0.8 g / L TSPG respectively. The cell viability was determined by MTT colorimetric method. Wright’s stain and light microscope were used to observe the cell morphological changes. Flow cytometry was used to determine the changes of cell cycle distribution and cell apoptosis rate. Immunocytochemistry was used to detect the expression of β-catenin and NF-κB p65 in the cells. Semi-quantitative RT-PCR detection of β-catenin mRNA and NF-κB p65 mRNA levels in the cells. Results Ginseng total saponin TSPG in vitro KG1α proliferation significantly inhibited. In the concentration range of 0.1 ~ 0.8 g / L TSPG, the inhibitory effect was concentration-dependent, reached the half-cell inhibitory rate at the concentration of 0.4 g / L, and reached the peak at 48 h. After treated with 0.4 g / L TSPG for 48 h, the number of cells dispersed and decreased obviously; the cells were obviously differentiated or apoptotic, produced mitosis, apoptotic bodies and vacuoles in the cytoplasm; compared with the control group, The cell cycle distribution of KG1α had no significant change (P> 0.05). Compared with the control group, the apoptotic rate of KG1α group was significantly increased (P <0.05). Compared with the control group, Compared with the control group, the expression of β-catenin protein in the group of cells decreased from the nucleus to the cell membrane, and the expression of NF-κB p65 also significantly decreased from the nucleus to the cell membrane. Compared with the control group, The levels of β-catenin and NF-κB p65 mRNA decreased, with statistical significance (P <0.05). Conclusion TSPG can inhibit KG1α cell proliferation induced apoptosis, and the mechanism of TSPG inhibiting KG1α cell proliferation may be related to the inhibition of β-catenin and NF-κB p65 protein expression, and reduce the level of β-catenin and NF-κB p65 mRNA.