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目的:研究姬松茸粗多糖对急性早幼粒细胞白血病细胞株HL-60细胞增殖、凋亡及转录因子NF-κB活性的影响并探索其诱导凋亡机制。方法:培养HL-60细胞,用不同浓度(1,2,5,10 mg/mL)姬松茸粗多糖诱导48 h后,采用WST-1法测定细胞增殖抑制;流式细胞术测定凋亡;免疫荧光法测定NF-κB P65蛋白核转运。结果:经不同浓度姬松茸粗多糖作用后,HL-60细胞存活率分别为90.0%,77.5%,47.5%,17.5%,与对照组(99.0%)比有统计学意义(P<0.05);5 mg/mL姬松茸粗多糖诱导48 h后,HL-60细胞早期凋亡率为52.4%,较对照组(6.0%)明显增加(P<0.05);免疫荧光显示NF-κB核转运较对照组减少。结论:姬松茸粗多糖可抑制HL-60细胞增殖,呈浓度-依赖性;能诱导HL-60细胞凋亡;诱导凋亡的作用与抑制NF-κB通路异常激活相关。
Objective: To study the effects of crude polysaccharide of Agaricus blazei Murill on proliferation, apoptosis and activity of transcription factor NF-κB in acute promyelocytic leukemia cell line HL-60 and explore its mechanism of apoptosis induction. METHODS: HL-60 cells were cultured and induced for 48 h with different concentrations (1, 2, 5, 10 mg/mL) of Agaricus blazei. The inhibition of cell proliferation was measured by WST-1 assay and apoptosis was determined by flow cytometry. The nuclear translocation of NF-κB P65 protein was measured by immunofluorescence. Results: The survival rate of HL-60 cells was 90.0%, 77.5%, 47.5%, and 17.5%, respectively, after treatment with different concentrations of crude Agaricus blazei polysaccharides, which was statistically significant compared with the control group (99.0%) (P<0.05). After induction of 5 mg/mL crude Agaricus blazei polysaccharide, the early apoptosis rate of HL-60 cells was 52.4%, which was significantly higher than that of the control group (6.0%) (P<0.05). Immunofluorescence showed that the NF-κB nuclear translocation was comparable to the control. Group decreases. Conclusion: Agaricus blazei Maxim polysaccharide can inhibit the proliferation of HL-60 cells in a concentration-dependent manner and induce apoptosis of HL-60 cells. The induction of apoptosis is related to the inhibition of abnormal activation of NF-κB pathway.