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  

Experimental Study on Eqi Compound Prescription Induced Apoptosis of Acute Myelocytic Leukemia HL-60

作　　者：郝军[1] 陈彻[1] 陈龙[2] 

机构地区：[1]甘肃中医学院,甘肃兰州730000 [2]兰州大学医学院附属第一医院,甘肃兰州730000

出　　处：《中国中医药信息杂志》2007年第6期29-31,共3页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：甘肃省教育厅科技基金项目(X001-03)

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、96h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72h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(P<0.01),Caspase-3浓度明显高于对照组(P<0.01),Bcl-2基因表达明显低于对照组(P<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。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 Ca^2＋ 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. Bci-2 gene expression were measured by LSAB. Results The contained-herb serum could induce apoptosis of HL-60 cells. Intracellular Ca^2＋ concentration of treatment with Eqi was higher evidently than that of control （P〈0.01）. Caspase-3 gene expression of treatment with Eqi was higher evidently than that of control （P〈0.01）. Bcl-2 gene expression of treatment with Eqi was lower evidently than that of control （P〈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 Ca^2＋, Caspase-3 and down-regulating Bcl-2.

关 键 词：莪芪抗瘤方剂 白血病 CA SPASE-3 BCL-2 

分 类 号：R289.11[医药卫生—方剂学]