青蒿素基于PI3K/AKT信号通路对人急性髓系白血病细胞凋亡的作用机制  

Mechanism analysis of the effect of artemisinin on cell apoptosis of human acute myeloid leukemia based on PI3K/AKT signaling pathway

作　　者：王朦 贾国存[1] 成怡冰 王海军[1] 刘炜[2] 郭亚琼 WANG Menga;JIA Guo-cuna;CHENG Yi-binga;WANG Hai-juna;LIU Weib;GUO Ya-qionga(Emergency Department, Zhengzhou Children's Hospital, Zhengzhou University affiliated Children's Hospital and Henan Children's Hospital, Zhengzhou 450000, Henan Province;Department of Hematology,Children's Hospital Affiliated to Zhengzhou University,Henan Children' s Hospital,Zhengzhou Children' s Hospital,Zhengzhou 450000,China)

机构地区：[1]郑州大学附属儿童医院暨河南省儿童医院郑州儿童医院急诊科,河南郑州450000 [2]郑州大学附属儿童医院暨河南省儿童医院郑州儿童医院血液科,河南郑州450000

出　　处：《实用药物与临床》2019年第10期1014-1018,共5页Practical Pharmacy and Clinical Remedies

基　　金：河南省2017年科技发展计划(172102310526)

摘　　要：目的分析青蒿素基于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路对人急性髓系白血病(AML)细胞凋亡的作用机制。方法以44例AML患儿原代细胞及K562细胞株4株为研究样本,平均分为对照组、实验组(分别加入12.5、25.0、50.0μg/ml青蒿琥酯),细胞计数法观察72h内细胞生长趋势,采用流式细胞术检测不同浓度青蒿琥酯对细胞凋亡的影响,免疫印迹法检测细胞凋亡相关蛋白(PI3K、AKT、P-AKT、Bcl-2、Bax、caspase-3、PTEN)的表达。结果AML原代细胞、细胞株K56224h内细胞存活率≥80%,对照组AML原代细胞、细胞株K56248~72h细胞存活率仍≥70%,而实验组AML原代细胞、细胞株K562培养48h细胞存活率均降至50%以下,且明显低于对照组(P<0.05);青蒿琥酯呈浓度依赖性诱导人AML原代细胞及K562细胞株凋亡,且25.0μg/ml组、50.0μg/ml组的细胞存活率低于12.5μg/ml组及对照组(P<0.05),25.0μg/ml组、50.0μg/ml组的细胞存活率比较差异无统计学意义(P>0.05)。培养48h后,实验组原代细胞及K562细胞株的PI3K、P-AKT、Bcl-2均明显下调(P<0.05),而Bax、caspase-3、PTEN表达上调(P<0.05),且青蒿琥酯浓度为25.0μg/ml组各项指标变化最明显,AKT无明显变化(P>0.05)。结论青蒿素类衍生物青蒿琥酯可经PI3K/AKT信号通路调节其Bcl-2、Bax、caspase-3、PTEN等凋亡相关蛋白表达,促进人AML细胞凋亡,25μg/ml浓度时效果较佳。Objective To analyze the mechanism of the effect of artemisinin on cell apoptosis of human acute myeloid leukemia ( AML) based on phosphatidylinositol 3-Kinases ( PI3K)/protein kinase ( AKT) signaling pathway. Methods The AML primary cells of 44 AML pediatic patients and 4 strains of K562 cell lines were enrolled as study samples. They were averagely divided into control group ( artesunate 0 μg /ml was given) and experimental group ( 12. 5, 25. 0 and 50. 0 μg /ml artesunate was given, respectively). The cell counting method was used to observe the cell growth trend within 72 h. The flow cytometry was used to detect the effect of artesunate at different concentrations on apoptosis. Western blot was used to detect the expression of apoptosis-related proteins ( PI3K,AKT,P-AKT,Bcl-2, Bax, caspase-3 and PTEN). Results The survival rate of AML primary cells and K562 cell lines within 24 h equaled to or was greater than 80%. The survival rate of AML primary cells and K562 cell lines within 48 ~ 72 h in control group still equaled to or was greater than 70%,while the survival rate of the above cells being cultured for 48 h in experimental group decreased to less than 50%,which was significantly lower than that in control group ( P < 0. 05). Artesunate induced apoptosis of human AML primary cells and K562 cell lines in a concentration-dependent way. The cell survival rates in the groups with 25. 0 μg /ml and 50. 0 μg /ml of artesunate were lower than that in the group with 12. 5 μg /ml of artesunate and control group ( P < 0. 05). There was no significant difference in cell survival rate between the groups with 25. 0 μg /ml and with 50. 0 μg /ml of artesunate ( P > 0. 05). After being cultured for 48 h, the expression of PI3K,PAKT and Bcl-2 of the primary cells and K562 cell lines in experimental group was significantly down-regulated ( P < 0. 05),while the expression of Bax, caspase-3 and PTEN was up-regulated ( P < 0. 05). The changes of each index was the most obvious in the group with 25. 0 μg /ml of artesunat

关 键 词：PI3K/AKT 信号通路 青蒿素 人急性髓系白血病 凋亡 

分 类 号：R285[医药卫生—中药学]