【摘要】 目的观察8-硝基白杨素(NOChR)抑制人急性髓性白血病细胞系(HL-60)细胞增殖和诱导凋亡的作用,并探讨其作用机制。方法体外培养HL-60细胞;MTT比色法检测增殖活性;流式细胞术定量分析细胞凋亡程度;琼脂糖凝胶电泳观察DNA梯形条带。Western Blot分析HL-60细胞PTEN、P-Akt蛋白表达的影响。结果NOChR可显著抑制HL-60细胞增殖,呈浓度依赖性,IC50为1.34μmol;NOChR具有诱导HL-60细胞凋亡的作用,且呈浓度和时间依赖性。NOChR以浓度和时间依赖方式上调HL-60细胞PTEN蛋白的表达和抑制P-Akt蛋白的表达,其作用可被PPARγ特异性阻断剂GW9662(10μmol)预孵育拮抗。结论NOChR诱导HL-60细胞凋亡作用与其活化PPARγ、上调PTEN蛋白表达、抑制Akt磷酸化相关。更多还原

【Abstract】 OBJECTIVE To observe the inhibition effect of 8-nitrochrysin on the proliferation of HL-60 cell and induction of HL-60 cell apoptosis,and investigate the action mechnism.METHODS HL-60 cell line was cultured in vitro and the proliferative activity of HL-60 cells treated with 5,7-dihydrox-8-nitrochrysin was measured using 3-(4,5-dimethylthiazol –2-yl)-2,5-diphenytertrazolium blue(MTT) colorimetric assay.The apoptosis rate of HL-60 cell line induced by NOChR was analyzed using flow cytometry(FCM) with PI staining.DNA ladder bands were observed by DNA agarose gel electrophoresis.The influence of NOChR on the PTEN and p-AKt protein expression of HL-60 cells was analyzed with western blot.RESULTS The proliferation of HL-60 cells was markedly inhibital by NOChR in a concentration-dependent manner,and the IC50 was 1.34 μmol.Flow cytometry after PI staining indicated that apoptosis in HL-60 cells was induced by NOChR in a time and concentration dependent manner.Western blot indicated that NOChR can efficiently up-regulated PTEN and down-regulated p-AKt protein expression of HL-60 cells in a concentration and time dependent manner,and the pre-incubation of GW9662,blocker of PPARγ,can antagonize the up-regulating effect of 8-nitrochrysin on PTEN protein expression of HL-60 cells.CONCLUSION NOChR induced apoptosis of HL-60 cells may be associated with activation of PPARγ,up-regulation of PTEN and inhibition of p-AKt by 8-nitrochyrsin.更多还原