别欧前胡内酯对HL-60细胞凋亡的影响

Influence of Alloimperatorin on the apoptosis of HL-60 cells

目的探讨别欧前胡内酯对人急性早幼粒细胞白血病HL-60细胞凋亡的影响及可能的机制。方法对数生长期人急性早幼粒细胞白血病HL-60细胞随机分为对照组和4、16、32、64、128mg/L别欧前胡内酯组。别欧前胡内酯作用24h,应用透射电子显微镜观察HL-60细胞形态学改变,MTT法检测别欧前胡内酯对HL-60细胞增殖的影响,根据细胞生长抑制率计算别欧前胡内酯对HL-60细胞的半数抑制浓度(half inhibitory concentration,IC50),流式细胞仪检测HL-60细胞凋亡率及线粒体膜电位改变。结果别欧前胡内酯作用24h,对照组HL-60细胞表面光滑,形态正常,大小均匀;64、128mg/L别欧前胡内酯组HL-60细胞数目明显减少,且体积变小,呈凋亡形态改变;MTT检测结果显示,别欧前胡内酯对HL-60细胞增殖具有明显的抑制作用,IC50值为(75±13)mg/L;流式细胞仪检测结果显示,对照组HL-60细胞凋亡率为(11.6±5.2)%,4、16、32、64、128 mg/L别欧前胡内酯组HL-60细胞凋亡率分别为(17.2±6.1)%、(24.2±8.6)%、(36.1±4.5)%、(46.0±7.3)%和(69.9±10.2)%,随别欧前胡内酯浓度增加,HL-60细胞凋亡率逐渐增高,线粒体膜电位逐渐降低(P<0.05)。结论别欧前胡内酯可明显促进人急性早幼粒细胞白血病HL-60细胞凋亡,其机制可能与改变HL-60细胞线粒体膜电位有关。

Objective To explore the influence and mechanism of Alloimperatorin on the apoptosis of human acute promyelocytic leukemia HL-60 ceils. Methods HL-60 ceils in logarithmic growth phase were randomly divided into control group, and 4, 16, 32, 64 and 128 mg/L Alloimperatorin groups. After the treatment with Alloimperatorin for 24 h, the morphological changes of HL-60 cells were observed under transmission electron microscopy. MTT assay was used to determine the influences of Alloimperatorin on the proliferation of HL-60 ceils, and the half inhibitory concentration （IC50） of Alloimperatorin on the proliferation of HL-60 cells was calculated according to the cell growth inhibition rate. The apoptosis rate and the change of mitochondrial membrane potential of HL-60 ceils were detected by flow cytometer. Results After the treatment with Alloimperatorin for 24 h, HL-60 cells in control group had a smooth surface, normal morphology and uniform size. In 64 and 128 mg/L Alloimperatorin groups, the number of HL-60 cells decreased significantly and the size became smaller, showing apoptosis morphological change. The results of MTT assay showed that Alloimperatorin had a significant inhibitory effect on the proliferation of HL-60 cells with the ICs0 value of （75±13） mg/L. The results of flow cytometry showed that the apoptosis rates of HL-60 cells were （11. 6 ±5. 2）%, （17.2±6.1） %, （24.2±8.6） %, （36.1±4.5）%, （46.0±7.3）% and （69.9±10.2）% in control group, and in 4, 16, 32, 64 and 128 mg/L Alloimperatorin groups, respectively, indicating that the apoptosis rate of HL-60 cells increased gradually and the mitochondrial membrane potential decreased with the increase of Alloimperatorin concentration （P〈 0.05）. Conclusion Alloimperatorin could obviously inhibit the apoptosis of HL-60 cells, and its mechanism may be correlated with the changing of the mitochondrial membrane potential of HL-60 cells.