纳米雄黄诱导白血病细胞药物敏感性的凋亡效应

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

作者信息

Wang Yong-Sheng, Zhou Si-Tong, Wei Hu-Lai

机构信息

Traditional Chinese Medicine Research Institute of Gansu Province, Central Laboratory, Lanzhou 730050, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2013 Jul;38(13):2202-5.

PMID:24079254

Abstract

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 leukemia 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 employed 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 microscopes. 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 x L(-1). After exposure to 20 mg x L(-1) and 50 mg x L(-1) nano-realgar for 48 hours, the apoptosis of K562 cells 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 x L(-1) and 50 mg x L(-1) nano-realgar for 48 h, the expression of P-53, Bax, Bcl-2 markedly increased in a time and dose-dependent manner. After administration of 20 mg x L(-1) and 50 mg x 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.

CONCLUSION

Nano-Realgar significantly induced apoptosis of drug-sensitive leukemia cells.

摘要

目的

探讨纳米雄黄对药物敏感白血病细胞的诱导凋亡作用。

方法

采用高能行星球磨机机械研磨制备纳米雄黄。以药物敏感白血病细胞（K562）为靶细胞，采用MTT法检测K562细胞的增殖活性，通过流式细胞术用FITC-Annexin V和碘化丙啶（PI）双染法研究细胞凋亡情况。采用流式细胞术（FCM）检测细胞内Bax、Bcl-2、P-53蛋白表达及Caspase-3活性。

结果

通过球磨将雄黄原药制成超细粉末，用电子显微镜测得纳米颗粒的平均直径为（72.72±22.18）nm。纳米雄黄显著抑制K562细胞增殖，处理24、48和72小时后，50%抑制浓度（IC50）分别为43.48、20.52、16.07mg·L⁻¹。经20mg·L⁻¹和50mg·L⁻¹纳米雄黄处理48小时后，Annexin V/PI染色检测的K562细胞凋亡增加，K562细胞凋亡率分别为10.52%和73.25%。靶细胞经20mg·L⁻¹和50mg·L⁻¹纳米雄黄处理48小时后，P-53、Bax、Bcl-2的表达呈时间和剂量依赖性显著增加。经20mg·L⁻¹和50mg·L⁻¹纳米雄黄处理48小时后，K562细胞中BCRP⁺、P-gp⁺以及同时表达P-gp和BCRP的细胞群体百分比显著增加。