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和厚朴酚对人白血病细胞系U937细胞增殖和凋亡的影响
Effects of Honokiol on Proliferation and Apoptosis on U937 Cells
【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。
【Abstract】 【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.
【Key words】 Honokiol; U937 cell line; membrane potential,mitochondrial; Caspase3/7;
- 【文献出处】 中山大学学报(医学科学版) ,Journal of Sun Yat-Sen University(Medical Sciences) , 编辑部邮箱 ,2009年04期
- 【分类号】R285.5
- 【被引频次】7
- 【下载频次】278