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目的:观察沙苑子总黄酮(FAC)对人急性早幼粒白血病细胞株HL-60的增殖抑制作用、诱导凋亡作用以及对p53蛋白表达的影响。方法:用SRB法检测不同浓度FAC作用于HL-60不同时间后的增殖抑制作用;选择作用较强的浓度处理HL-60细胞,在不同时间点收集细胞,用流式细胞术和Hoechst荧光染色检测细胞凋亡;免疫荧光法检测HL-60细胞p53蛋白含量。结果:FAC对HL-60有明显的增殖抑制作用,在浓度为0.25 mg/ml作用24h、36h、48h后以及0.125 mg/ml与0.063mg/ml作用36h、48h后对HL-60都有明显的抑制作用。在0.63mg/ml、0.125mg/ml、0.25mg/ml浓度FAC作用36h后,其细胞凋亡率分别为38.3%、49.6%、57.9%,FAC诱导细胞凋亡具有剂量-效应关系和时间-效应关系。经Hoechst染色可见细胞染色质固缩,细胞膜完整等凋亡改变。免疫荧光检测HL-60细胞p53蛋白表达增高。结论:FAC具有抑制HL-60增殖作用,其作用在一定范围内呈剂量-效应关系以及时间-效应关系,其作用机制可能是通过提高p53蛋白表达诱导HL-60细胞凋亡。
Objective: To observe the inhibitory effect of FAC on the proliferation of human acute promyelocytic leukemia cell line HL-60, the induction of apoptosis and the expression of p53 protein. Methods: The proliferation inhibition of HL-60 cells treated with different concentration of FAC was detected by SRB method. HL-60 cells were treated with strong concentration and collected at different time points. Flow cytometry and Hoechst staining The apoptosis of HL-60 cells was detected by immunofluorescence. RESULTS: FAC had a significant inhibitory effect on the proliferation of HL-60 cells. After treated with 0.25 mg / ml FAC for 24 h, 36 h, 48 h and 0.125 mg / ml and 0.063 mg / ml for 36 h and 48 h, Inhibition. The apoptotic rates of FAC induced by FAC were 38.3%, 49.6% and 57.9% at the concentrations of 0.63mg / ml, 0.125mg / ml and 0.25mg / ml for 36h, respectively. There was dose-effect relationship and time- Effect relationship. Hoechst staining showed chromatin condensation, cell membrane integrity and other apoptosis changes. Immunofluorescence detected HL-60 cells increased p53 protein expression. CONCLUSION: FAC can inhibit the proliferation of HL-60 cells in a dose-dependent and time-effect manner. The possible mechanism is that FAC can induce the apoptosis of HL-60 cells by increasing the expression of p53 protein.