摘要
目的观察丹参酮Ⅱa对HL-60的凋亡与自噬以及相关因子水平的影响,探讨丹参酮Ⅱa抗急性髓系白血病的分子机制。方法使用不同浓度的丹参酮Ⅱa处理HL-60细胞24、48 h后用CCK-8法检测增殖抑制率,吉姆萨染色观察细胞形态,Western blotting检测Caspase-3、Cleaved-Caspase-3、PARP-1、p-AMPK、pmTOR、LC3B水平变化,联用自噬抑制剂巴弗洛霉素A1以CCK-8测细胞存活率。结果以0、1、2、4、8、16、32、64、128μmol/L丹参酮Ⅱa处理HL-60细胞24、48 h后HL-60增殖被抑制,其作用随浓度增加和时间延长而加强,24、48 h的IC_(50)分别为32.87、18.4μmol/L;吉姆萨染色镜下观察见,随着药物浓度增加HL-60凋亡增加;Western blotting显示Caspase-3减少、Cleaved-Caspase-3增加,PARP-1切割增加,凋亡增强;p-AMPK增加、pmTOR减少、LC3B增加,显示AMPK/mTOR自噬通路激活,联用巴弗洛霉素A1后丹参酮Ⅱa增殖抑制作用减弱。结论丹参酮Ⅱa通过诱导HL-60凋亡和自噬发挥抗白血病作用,AMPK/mTOR可能是介导这一效应的重要信号通路。
Objective:To observe the effects of tanshinoneⅡa on apoptosis and autophagy of acute myeloid leukemia(AML)cell HL-60 cells,as well as relative factors,to investigate the molecular mechanism of tanshinoneⅡa against AML.Methods:HL-60 cells were treated with different concentrations of tanshinoneⅡa for 24,48 h,and then CCK-8 method was used to measure the changes in the rate of proliferation inhibition and IC50.Changes in Caspase-3,PARP-1,p-AMPK,p-mTOR,LC3 B levels were measured by western blotting,and cell survival was measured by CCK-8 in combination with the autophagy inhibitor bafilomycin A1.Result:HL-60 cell proliferation was inhibited after treatment of HL-60 cells with 0,1,2,4,8,16,32,64,128μmol/L tanshinoneⅡa for 24,48 h,and its effect intensified with increasing concentration and time,with IC50 of 32.87μmol/L and 18.4μmol/L for 24 h and 48 h,respectively.IC_(50)was used as the basis to set up the control group and the low,medium and high dose groups,and it was observed under the microscope that HL-60 apoptosis increased as the concentration increased,and Western blotting showed that Caspase-3 decreased,Cleaved-Caspase-3 increased,PARP-1 cut increased,showing its apoptosis enhanced,p-AMPK increased,p-mTOR decreased,LC3 B increased,showing activation of the AMPK/mTOR autophagy pathway,and decreased proliferation inhibition of TanshinoneⅡa after coadministration of bafilomycin A1.Conclusion:TanshinoneⅡa exerts anti-leukemic effects through induction of HL-60 apoptosis and autophagy,and AMPK/mTOR may be an important signaling pathway mediating this effect.
作者
潘一鸣
李蕊白
黄子明
王婧
马薇
陈信义
侯丽
Pan Yiming;Li Ruibai;Huang Ziming;Wang Jing;Ma Wei;Chen Xinjd;Hou Li(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)
出处
《中国中医急症》
2021年第3期415-418,共4页
Journal of Emergency in Traditional Chinese Medicine
基金
国家自然科学基金项目(82074240)
国家重点研发计划项目(2016YFE0202800)。
