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.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
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)
