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作 者:薛芳[1] 成志勇[2] 杨琳[1] 李世辉[3] 张敬宇[1] 姚丽[1] 潘峻[1]
机构地区:[1]河北医科大学第二医院血液内科,河北省血液病重点实验室,河北石家庄050000 [2]保定市第一医院血液科,河北保定071000 [3]北京航空工业中心医院肿瘤科,北京100012
出 处:《中山大学学报(医学科学版)》2009年第4期408-412,共5页Journal of Sun Yat-Sen University:Medical Sciences
基 金:河北省科技攻关计划项目(072761130)
摘 要:【目的】探讨和厚朴酚(HNK)对白血病细胞系U937细胞增殖和凋亡的影响。【方法】将U937细胞和正常外周血单核细胞分别与HNK共培养,Hoechst33342荧光染色检测细胞凋亡形态,用噻唑蓝比色法(MTT)检测HNK对细胞增殖的抑制作用;流式细胞仪检测凋亡;罗丹明123检测线粒体膜电位;Caspase3/7活性检测试剂盒检测Caspase3/7活性;荧光定量PCR(FQ-PCR)检测Caspase-3、Caspase-7mRNA水平变化。【结果】HNK对U937的体外增殖有显著抑制作用,48h半数抑制浓度(IC50)为11.8μg/mL,而对正常外周血单核细胞的IC50为40.3μg/mL,AnnexinV/PI双标染色显示,细胞凋亡与和厚朴酚呈浓度和时间依赖相关。HNK明显降低U937细胞线粒体膜电位,增加Caspase3/7活性及mRNA表达水平,但对正常外周血单核细胞的增殖抑制和凋亡作用不明显。【结论】HNK可能通过激活caspase-3/7抑制U937细胞增殖、诱导U937细胞凋亡。[Objective] To investigate the anti-proliferative and apoptosis effect induced by Honokiol (HNK) on human myeloid leukemia cell line U937 cells in vitro. [ Methods ] After treated with different concentration of HNK, Hoechst33342 fluorescent staining was used to detect cell apoptosis; the growth inhibition ration of U937 cells and PBMCs were analyzed by MTT assay; the apoptosis ration was detected by flow cytometry; mitochondrial membrane potential was explored by rhodamine 123 stain; Caspase3/7 protein activity kit was used to test the Caspase3/7 activity; the Caspase-3 and Caspase-7 mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR (FQ-PCR). [ Results ] Honokiol could significantly inhibit the proliferation of U937 cells in terms of the indexes of IC50/U937 11.8 μg/mL and IC50/PBMCs 40.3 μg/ mL, and the anti-proliferative effect was in a time and concentration dependent manner; Flow cytometry analysis manifested that Honokiol could induce U937cells apoptosis by Annexin V/PI double Annexin V/PI fluorescein stain; Honokiol significantly inhibited the mitochondrial membrane potential of U937 cells and enhanced the ability of Caspase3/7 and the mRNA expression levels, but not the PBMCs. [ Conclusion] HNK can inhibit U937 cells proliferation and induce cells apoptosis via activating Caspase 3/7.
关 键 词:和厚朴酚 U937细胞系 膜电位 线粒体 Caspase3/7
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