青蒿琥酯对K562细胞和脐血CD34^+造血干/祖细胞的选择性作用  

Selective effect of artesunate on K562 cell and CD34^+ hematopoietic stem/progenitor cell from umbilical cord blood

作　　者：林雪梅[1] 田雪莲[2] 杨慧[1] 宋姝丹[1] 王莎莉[2] 王亚平[1] 

机构地区：[1]重庆医科大学基础医学院组织学与胚胎学教研室 [2]重庆医科大学基础医学院生理学教研室

出　　处：《第三军医大学学报》2007年第20期1939-1942,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30472253);重庆市科委课题(渝科发计字2005-50)~~

摘　　要：目的探讨青蒿琥酯(artesunate,AST)对K562细胞和正常造血干/祖细胞是否具有选择性作用。方法选用K562细胞及脐血CD34+造血干/祖细胞,加入不同浓度AST。MTT法检测药物对上述两种细胞增殖的影响;甲基纤维素半固体培养法考察AST对其体外扩增和多向分化能力的影响;以流式细胞术Annexin V-FITC-PI双染定量检测细胞凋亡率;Western blot方法检测BCR-ABL融合蛋白的表达。结果青蒿琥酯对K562细胞有明显的抑制增殖和诱导凋亡的作用(P<0.01),并存在时效和量效关系,对集落抑制作用明显(P<0.01)。青蒿琥酯在12.5~25μg/ml作用48h对脐血CD34+造血干/祖细胞增殖抑制作用和诱导凋亡作用不明显(P>0.05),对集落生成亦无明显抑制作用(P>0.05)。经青蒿琥酯作用48h,K562细胞BCR-ABL蛋白以剂量依赖方式被降解(P<0.01)。结论青蒿琥酯在一定浓度范围内对K562细胞具有选择性抑制增殖和诱导凋亡的作用,对脐血CD34+造血干/祖细胞影响较小。Objective To study the selective effect of artesunate （AST） on K562 cells and the normal hematopoietic stem/progenitor cells. Methods K562 cells and the CD34^＋ hematopoietic stem/progenitor cell （HSC/HPC） from umbilical cord blood were respectively cocultured with AST of different concentrations （50, 25, 12.5 μg/ml）. The inhibition rate of the cellular growth was measured by MTT assay. The capability of proliferation and multilineage differentiation was studied by methylcellulose assay systems. Stained by anti Annexin V-FITC and PI, their apoptotic rates were examined by flow cytometer. The expressions of BCR-ABL protein were analyzed by Western blot. Results Artesunate had a significant inhibition and apoptosis-inducing effects on K562 cells in both time- and dose-dependent manners （P 〈 0. 01 ）. The number of CFCs was also markedly decreased （P 〈0. 01 ）. However, the treatment of artesunate at dose of 12.5, 25 μg/ml for 48 h, had little inhibition and apoptosis-inducing effects on hematopoietic stem/progenitor cells （P 〉 0. 05 ）. The number of CFCs was not affected, either. The expression of BCR-ABL protein could be down-regulated in a dose-dependent manner after 48-hour treatment of AST （P 〈 0. 01 ）. Conclusion The inhibition and apoptosis-inducing effects of artesunate are specific to K562 cells, while AST at dose of 12.5,25 μg/ml hardly affected HSC/HPC.

关 键 词：青蒿琥酯 K562细胞 CD34^+造血干/祖细胞 增殖 分化 凋亡 

分 类 号：R285.5[医药卫生—中药学] R73-354[医药卫生—中医学]