纳米雄黄对药物敏感性白血病细胞的凋亡诱导作用  

Apoptosis effects of drug sensitivity leukemia cells induced by nano-realgar

作　　者：王永胜[1] 周思彤[2] 魏虎来[3] 

机构地区：[1]甘肃省中医药研究院中心实验室,甘肃兰州730050 [2]甘肃省中医院检验科,甘肃兰州730000 [3]兰州大学基础医学院甘肃省新药临床前研究重点实验室,甘肃兰州730000

出　　处：《中国中药杂志》2013年第13期2202-2205,共4页China Journal of Chinese Materia Medica

基　　金：甘肃省中医药科学技术研究课题(GZK-2012-17);甘肃省自然科学研究基金计划项目(096RJZA034)

摘　　要：目的:研究纳米雄黄对药物敏感性白血病细胞的凋亡诱导作用。方法:采用机械研磨法制备纳米雄黄。以白血病药物敏感K562细胞为靶细胞,采用MTT法检测纳米雄黄对K562细胞的增殖抑制作用,AnnexinV/PI双染色法检测细胞凋亡,细胞流式细胞术检测Bax,Bcl-2,P-gp,BCRP,P-53蛋白的表达水平,Caspase-3活性。结果:雄黄经高能球磨机球磨10～50h,扫描电镜(SEM)和透射电镜(TEM)观察,电镜测量纳米雄黄的平均粒径为(72.72±22.18)nm,符合纳米颗粒的形貌特征。纳米雄黄对K562细胞有明显的增殖抑制作用。24,48,72 h的IC50分别为43.48,20.52,16.07 mg.L-1。20,50 mg.L-1纳米雄黄作用48 h后Annexin V/PI染色显示凋亡细胞明显增高,凋亡率分别为10.52%,73.25%。纳米雄黄作用48 h后Bax蛋白表达由对照的(75.80±2.40)%升高到(87.50±1.20)%和(99.60±0.10)%;Bcl-2蛋白表达由对照的(73.85±0.15)%升高到(94.80±2.00)%和(97.75±0.55)%。20,50 mg.L-1纳米雄黄处理K562细胞24 h后,K562细胞Caspase-3蛋白表达水平由对照的(3.14±0.24)%升高到(8.87±2.74)%和(72.55±1.45)%;细胞的BCRP,P-gp蛋白,P53蛋白表达总体呈上升趋势,共表达也呈上升趋势。结论:纳米雄黄可以显著诱导白血病药物敏感K562细胞发生凋亡。Objective： To explore apoptosis-inducing effects of realgar nanoparticle （nano-realgar） on drug-sensitive leukemia cells. Method ： Preparation of nano-realgar was mechanical milled using a high-energy planetary ball mill. Using drug-sensitive leuke- mia cells（K562） as target cells, MTT assay was used to detect the proliferating activity of K562 cells, and the cellular apoptosis was investigated with double staining of FITC-Annexin V and propidium iodide （PI） by flow cytometry. Flow cytometry （FCM） was em- ployed to detect expression of intracellular Bax, Bcl-2, P-53 protein and the activity of Caspase-3. Result： The raw realgar was made to ultra-fine powder by ball milling, and the average diameter of the nanoparticle was （72. 72±22. 18） nm measured with electron mi- croscopes. Nano-realgar significantly inhibited the proliferation of K562 cells, Treated for 24, 48 and 72 hours, the 50% inhibitory concentration （ IC50 ） was 43.48, 20. 52,16. 07 mg. L-1. After exposure to 20 mg . L-1 and 50 mg . L-1 nano-realgar for 48 hours, the apoptosis of K562 ceils detected by Annexin V/PI staining was increased, the apoptotic rate of K562 cells was 10. 52% and 73.25%. After the target cells were treated with 20 mg . L-1 and 50 mg .L-1 nano-realgar for48 h, the expression of P-53,Bax,Bcl- 2 markedly increased in a time and dose-dependent manner. After administration of 20 mg. L- 1 and 50 mg . L- 1 nano-realgar for 48 h, the percentage of BCRP ＋ , P-gp＋ and co-expressing P-gp and BCRP cell population in K562 cells incrased dramatically. Conclu- sion： Nano-Realgar significantly induced apoptosis of drug-sensitive leukemia cells.

关 键 词：纳米雄黄 白血病 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]