益气养阴法方药促进KG1a进入细胞周期而增强DNR敏感性的研究

Study on Supplementing qi and Nourishing Yin Prescription of Promoting KG1a to Enter Cell Cycle and Enhancement of DNR Sensitivity

目的:探讨益气养阴法方药对白血病干细胞(Leukemia Stem Cells,LSC)KG1a细胞周期及其增殖的影响。方法:WST-8法检测益气养阴方含药血清对KG1a体外增殖的影响;WST-8法检测经益气养阴方含药血清联合柔红霉素(DNR)对的KG1a细胞体外增殖的影响;流式细胞术分析益气养阴方含药血清对KG1a细胞周期G0/G1期、S期、G2M期的影响。结果:益气养阴方含药血清对KG1a细胞体外增殖抑制作用不明显,呈一定时间及剂量依赖性,但各浓度相比抑制率无明显差异;含药兔血清20μL、40μL浓度组联合DNR对KG1a细胞有明显的抑制作用,且随浓度的增加和作用时间的延长而增强,且40μL浓度组含药兔血清联合DNR组48 h(72.5%)后抑制率显著高于DNR组(P<0.01);益气养阴方含药血清可促使KG1a细胞由G0期进入细胞周期,并呈剂量依赖性;结论:益气养阴含药血清可能通过促进KG1a细胞由G0期进入细胞周期,从而增强DNR的体外杀伤作用,可显著降低白血病细胞的耐药性。

Objective： To investigate the effect of supplementing qi and nourishing Yin recipe serum on leukemia stem cells（ LSC） KG1 a cell cycle and proliferation in vitro. Method： The inhibition effects of proliferation were assayed by WST- 8 method in KG1 a cells treated by supplementing qi and nourishing Yin prescription for 24,48,72 hours. WST-8 method was used to detect the effect of supplementing qi and nourishing Yin recipe serum combined with daunorubicin（ DNR） on the proliferation of KG1 a cells in vitro. We analyzed the effect of supplementing qi and nourishing Yin recipe serum on cell cycle of KG1 a G0 / G1 phase,S phase,G2 M phase by flow cytometry. Result： The inhibitory effect of supplementing qi and nourishing Yin recipe serum on the proliferation of KG1 a cells in vitro was not obvious,which with a certain dose- and time- dependent manner,but the inhibitory rate compared to the concentration had no significant difference. The rabbit serum concentration groups of 20 μL and 40 μL combined with DNR can significantly inhibit the proliferation of KG1 a cells,prolonged with the increase of concentration and time. And the inhibitory rate of 40 μL concentration group containing rabbit serum plus DNR group after 48 h（ 72. 5%） was higher than that in DNR group（ P〈0. 01）.The supplementing qi and nourishing Yin recipe serum can induce KG1 a cells from G0 phase to enter the cell cycle with a certain dose- dependent manner. Conclusion： Supplementing qi and nourishing Yin recipe serum can promote KG1 a cells into the cell cycle from G0 phase,which can significantly reduce the drug resistance of leukemia cells,and thus enhance the cytotoxicity of DNR in vitro.