莪芪抗瘤方剂you导白血病HL-60细胞凋亡的血清药理学研究.docVIP

　　莪芪抗瘤方剂诱导白血病HL-60细胞凋亡的血清药理学研究 【摘要】 目的 研究中药复方莪芪抗瘤方剂在体外对白血病HL-60细胞凋亡的作用,并探讨凋亡发生与细胞内Ca2+、Caspase-3、Bcl-2的关系。方法 通过血清药理学方法,选择10%浓度的含药血清与白血病HL-60细胞共同培育24、48、72、96 h后,应用荧光显微镜观察细胞形态,流式细胞仪检测凋亡峰(亚二倍体峰),用Fura-2荧光负载方法测定细胞内Ca2+浓度的变化。比色法检测凋亡相关基因Caspase-3活性变化。亲和免疫组化LSAB 法检测HL-60细胞Bcl-2基因表达。结果 10%浓度的含药血清处理组细胞体积缩小,细胞核固缩,膜起泡,核断裂及形成凋亡小体等,以72 h最为明显；可见DNA含量低于G1期的凋亡细胞群,这群细胞的凋亡百分率随时间的递增而增加。10%浓度的含药血清处理组的细胞内Ca2+浓度也明显高于对照组(Plt;0.01),Caspase-3浓度明显高于对照组(Plt;0.01),Bcl-2基因表达明显低于对照组(Plt;0.05)。结论 莪芪抗瘤方剂含药血清诱导白血病HL-60细胞发生凋亡,这种变化可能与细胞内Ca2+、Caspase-3水平上调及Bcl-2表达下调有关。 【关键词】 莪芪抗瘤方剂；白血病；Caspase-3；Bcl-2 　　Abstract：Objective To explore the apoptotic effect of Eqi compound prescription (contained-herb serum) on cute myelocytic leukemia HL-60 cell, which is relative to intracellular Ca2+, Caspase-3 and Bcl-2. Methods According to serum pharmacology, HL-60 cells were exposed to 10% concentrations of contained-herb serum for 24, 48, 72 and 96 hours respectively. Cells were observed under a fluore- scence microscope. SubG1 DNA was examined by flow cytometer. Intracellular Ca2+ concentration was measured by fura-2 fluorescence load method. Caspase-3 enzymatic activity were measured by colorimetry. Bcl-2 gene expression were measured by LSAB. Results The contained-herb serum could induce apoptosis of HL-60 cells. Intracellular Ca2+ concentration of treatment with Eqi was higher evidently than that of control (Plt;0.01). Caspase-3 gene expression of treatment with Eqi was higher evidently than that of control (Plt;0.01). Bcl-2 gene expression of treatment with Eqi was lower evidently than that of control (Plt;0.05). Conclusion According to serum pharmacology, Eqi compound prescription could induced apoptosis of acute myelocytic leukemia HL-60 cell, which is relative to up-regulating Ca2+, Caspase-3 and down-regulating Bcl-2. 　　Key words：Eqi compound prescription；acute myelocytic leukemia；apoptosis；Caspase-3；Bcl-2　　越来越多的研究表明,细胞内Ca2+水平升高与诱发细胞凋亡密切相关[1]。细胞内凋亡相关基因Caspase-3活性变化对凋亡的诱导起重要促进作用[2]。Bcl-2 是重要的抑凋亡基因之一,过度表达