Effect of Astragalus Polysaccharides on Proliferation,Apoptosis,and Cycle of Jurkat Cells in Leukemia
尹宝慧 1耿雪静 1姜吉祥 1李有杰 2孙允霄1
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作者信息
1. 滨州医学院烟台附属医院,烟台,264100
2. 滨州医学院,烟台,264003
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摘要
目的:研究黄芪多糖对白血病细胞(Jurkat)增殖、凋亡、周期的影响,并进一步探讨其作用机制.方法:不同浓度黄芪多糖(0、100、200、400 μg/mL)作用于Jurkat细胞,细胞计数(CCK-8)法检测各组细胞增殖抑制率,流式细胞术检测各组细胞周期及细胞凋亡情况,实时荧光定量PCR(Real Time PCR)法及蛋白印迹法检测细胞周期素D1(CyclinD1)、细胞周期素E(CyclinE)、胱天蛋白酶3(Caspase3)、胱天蛋白酶8(Caspase 8)的mRNA及蛋白表达.结果:不同浓度黄芪多糖(100、200、400 μg/mL)均有效抑制Jurkat细胞增殖,呈剂量及时间依赖性(P＜0.05);流式细胞术检测各组细胞周期及细胞凋亡情况,随黄芪多糖浓度增加,G0/G1期细胞逐渐增多,G2/M期细胞逐渐减少,细胞凋亡率逐渐增加,与对照组比较,差异有统计学意义(P＜0.05);黄芪多糖处理组细胞中CyclinD1、CyclinE mRNA及蛋白表达下降、Caspase3、Caspase8mRNA及蛋白表达升高,差异有统计学意义(P＜0.05).结论:黄芪多糖能有效抑制白血病Jurkat细胞增殖,促进其凋亡,发挥抗白血病作用.其作用机制与调控CyclinD1、CyclinE表达,阻滞细胞周期于G0/G1期,促进凋亡相关基因Caspase3、Caspase8表达有关.
Abstract
Objective:To investigate the effect of Astragalus polysaccharides on the proliferation,apoptosis,and cell cycle of leuke-mia cells(Jurkat),and to further explore its underlying mechanisms.Methods:Jurkat cells were treated with different concentra-tions of Astragalus polysaccharides(0,100,200,400 μg/mL).The cell counting kit-8(CCK-8)assay was used to detect the prolif-eration inhibition rate of each group.Flow cytometry was employed to assess cell cycle distribution and apoptosis.Real-time quanti-tative PCR(Real-Time PCR)and Western blot were used to detect the mRNA and protein expression of Cyclin D1,Cyclin E,Caspase 3,and Caspase 8.Results:Astragalus polysaccharides at concentrations of 100,200,and 400 μg/mL effectively inhibited Jurkat cell proliferation in a dose-and time-dependent manner(P＜0.05).Flow cytometry analysis showed that compared with the control group,with increasing concentrations of Astragalus polysaccharides,the number of cells in the G0/G1 phase increased,while the number of cells in the G2/M phase decreased,and the apoptosis rate gradually increased(P＜0.05).In the Astragalus polysac-charides-treated groups,the mRNA and protein expression of Cyclin D1 and Cyclin E decreased,while the mRNA and protein ex-pression of Caspase 3 and Caspase 8 increased(P＜0.05).Conclusion:Astragalus polysaccharides can effectively inhibit the pro-liferation of Jurkat leukemia cells and promote their apoptosis,exerting an anti-leukemia effect.The mechanism of action is related to the regulation of Cyclin D1 and Cyclin E expression,blocking the cell cycle at the G0/G1 phase,and promoting the expression of apoptosis-related genes Caspase 3 and Caspase 8.
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