摘要
目的:研究槲皮素对白血病细胞的杀伤作用以及诱导的凋亡研究。方法:CCK-8试剂盒检测不同浓度的槲皮素(0~100μM)对白血病K562和HL-60细胞活性的影响;Hoechst33342染色技术观察40u M槲皮素处理细胞24和48小时后细胞核形态的变化;流式细胞仪检测细胞凋亡情况;caspase8、9、3(半胱天冬酶)特异性抑制剂预处理K562细胞后,检测药物对细胞活性的影响;40u M槲皮素处理细胞24和48小时后,Western blot检测细胞中caspase家族蛋白8、9、3的表达情况。结果:0~100u M的槲皮素能够引起K562和HL-60细胞剂量依赖型的活性下降,40u M的槲皮素处理细胞24和48小时后,K562细胞核明显发生皱缩,引起凋亡,随着处理时间增长,凋亡率增加,处理24和48小时后细胞的凋亡率分别为(45.8±2.06)%和(83.6±1.89)%。caspase 9、3抑制剂预处理K562细胞后,槲皮素能够引起细胞活性的增加,而caspase 8抑制剂预处理后,槲皮素没有引起细胞活性的上升。Western blot检测结果表明40μM的槲皮素处理细胞后能够上调caspase 9、3蛋白的表达,而没有引起caspase 8蛋白表达量的上升,随着处理时间的增长,caspase 9、3的表达量更高。结论:槲皮素对白血病细胞具有很强的杀伤作用,并诱导细胞caspase 9、3蛋白参与的内源性通路介导的凋亡。
Objective: To investigate the killing effect of quercetin on leukemia cells and the cell apoptosis induced by quercetin. Methods: CCK-8 assays was used to detect the viability of leukemia cell K562 and HL-60 after treating with different concentration of quercetin( 0 ~ 100 u M); Hoechst33342 was used to stain the cell to observe the nucleus change; flow cytometry was further used to test the K562 cell apoptosis and its apoptotic ratio; after pretreatment with caspase8,9,3 inhibitors,the cell viability was detected; at last,western blot was used to detect the caspase8,9,3 protein expression of K562 cell after treating with 40 u M quercetin for 24 and 48 h. Results: 0 ~ 100 u M of quercetin could induce the dose-dependent viability decrement of K562 and HL-60 cells( p 0. 05). Compared with control group,after treating with40 u M of quercetin for 24 and 48 h,K562 cells showed significant nucleus pyknosis and further apoptosis,which the apoptotic ratio was( 45.8 ± 2. 06) % and( 83. 6 ± 1. 89) %( p 0. 01) respectively. After pretreatment with caspase 9,3 inhibitors,quercetin could enhance the cell viability compared with single quercetin treating group( p 0. 01),but caspase 8 didn 't had similar effect. At last,western blot results showed that quercetin could up-regulate the caspase 9 and 3 protein expression but not caspase 8 protein. Conclusions: Caspase protein family members 9,3 joined the quercetin induced leukemia cell apoptosis. Quercetin had strong killing effect on leukemia cells and induced endogenous pathways mediated apoptosis.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2015年第5期61-65,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
河南省教育厅基金资助项目(编号:2006320039)