青蒿琥酯经10号染色体缺失的磷酸酶及张力蛋白同源基因通路诱导K562细胞凋亡的作用机制研究  

Study on the effective mechanisms of Artesunate inducing apoptosis on K562 cells through phosphatase and tensin hemology deleted on chromosome ten gene pathway

作　　者：成志勇[1] 颜晓燕[1] 李琳[1] 谢旭磊[1] 王凤云[1] 李继业[1] 王素云[2] 

机构地区：[1]保定市第一医院血液内科,河北保定071000 [2]河北省人民医院血液内科,石家庄050000

出　　处：《中国临床药理学杂志》2015年第11期938-941,共4页The Chinese Journal of Clinical Pharmacology

基　　金：河北省科技厅科技支撑计划基金资助项目(12276104D-1)

摘　　要：目的探讨青蒿琥酯经10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)介导的信号通路诱导人白血病K562细胞凋亡作用机制。方法将5个浓度(0,6.25,12.5,25,50μg·m L-1)浓度的青蒿琥酯作用K562细胞0-72h,通过噻唑蓝比色法观察其对K562细胞生长抑制作用;罗丹明123检测线粒体膜电位变化;荧光定量聚合酶链反应检测PTEN mRNA水平变化,试剂盒检测半胱天冬酶(Caspase)3/7活性;蛋白印迹法检测PTEN、蛋白激酶B(p-Akt)蛋白水平。结果青蒿琥酯能显著抑制K562细胞增殖,72 h最大抑制率为91.5%,与剂量和时间呈正相关。青蒿琥酯明显降低K562细胞线粒体膜电位,增加PTEN表达,抑制p-Akt表达,增强Caspase3/7蛋白活性。结论青蒿琥酯可能通过PTEN/Akt/Caspase通路诱导K562细胞凋亡。Objective To investigate the effective mechanism of Artesu-nate inducing apoptosis in human leukemia K562 cells through phospha-tase and tensin hemology deleted on chromosome ten gene （ PTEN） path-way.Methods The K562 cells were treated with concentrations of 0 , 6.25,12.5,25,50μg.mL^-1 Artesunate within 0-72 hours.The growth inhibition of Artesunate on K562 cells was detected by thiazole blue assay.The mitochondrial membrane potential was assessed by rhodamine 123 with flow cytometry.The PTEN mRNA expression levels were detec-ted by real-time fluorescent quantitative reverse transcription polymerase chain reaction.Cysteine aspartic acid specific protease （ Caspase ） -3/7 protein activitives were tested by kits.PTEN and protein kinase B （ p-Akt） protein levels weres detected by Western Blotting.Results Arte-sunate has significant proliferation inhibition effect on K562 cells.The maximum growth inhibition ratio was 91.5% in 72 hours and the anti-proliferative effect was in a time and dose dependent manner.The mito-chondrial membrane potential was significantly decreased.PTEN was up regulated.The p-Akt was down regulated and caspase-3/7 protein ac-tivity was enhanced by Artesunate.Conclusion Artesunate possiblely induced K562 cells apoptosis via PTEN/Akt/Caspase pathway.

关 键 词：青蒿琥酯 K562细胞系 10号染色体缺失的磷酸酶基因 凋亡 

分 类 号：R965.1[医药卫生—药理学] R735.7[医药卫生—药学]