三氧化二砷联合双氢青蒿素对THP-1细胞增殖及凋亡的影响

赵丽凤; 李晓强; 任汝静; 刘丽; 谭余庆

doi:10.13422/j.cnki.syfjx.20201737

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三氧化二砷联合双氢青蒿素对THP-1细胞增殖及凋亡的影响

药理 | 更新时间：2020-09-09

- 三氧化二砷联合双氢青蒿素对THP-1细胞增殖及凋亡的影响
- Effect of Arsenic Trioxide Combined with Dihydroartemisinin on THP-1 Cells Proliferation and Apoptosis
- 中国实验方剂学杂志 2020年26卷第17期页码：42-48
- 作者机构：
  
  中国中医科学院中药研究所，北京 100700
- 作者简介：
  
  赵丽凤，在读硕士，从事中医药抗白血病研究，E-mail：1585741203@qq.com
  刘丽，助理研究员，从事中医药抗白血病研究，E-mail：lliu1990@icmm.ac.cn
  谭余庆，研究员，从事中医药抗白血病研究，E-mail：yqtan@icmm.ac.cn
- 基金信息：
  
  中央级公益性科研院所基本科研业务费专项(ZZ13-YQ-054)
- DOI：10.13422/j.cnki.syfjx.20201737
  中图分类号： R2-0;R289;R733.7
- 网络出版日期：2020-06-30，
  
  纸质出版日期：2020-09-05
- 稿件说明：
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赵丽凤,李晓强,任汝静等.三氧化二砷联合双氢青蒿素对THP-1细胞增殖及凋亡的影响[J].中国实验方剂学杂志,2020,26(17):42-48.

ZHAO Li-feng,LI Xiao-qiang,REN Ru-jing,et al.Effect of Arsenic Trioxide Combined with Dihydroartemisinin on THP-1 Cells Proliferation and Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(17):42-48.
赵丽凤,李晓强,任汝静等.三氧化二砷联合双氢青蒿素对THP-1细胞增殖及凋亡的影响[J].中国实验方剂学杂志,2020,26(17):42-48. DOI： 10.13422/j.cnki.syfjx.20201737.

ZHAO Li-feng,LI Xiao-qiang,REN Ru-jing,et al.Effect of Arsenic Trioxide Combined with Dihydroartemisinin on THP-1 Cells Proliferation and Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(17):42-48. DOI： 10.13422/j.cnki.syfjx.20201737.

摘要

目的

研究三氧化二砷联合双氢青蒿素对人急性单核细胞白血病（AML）细胞株THP-1细胞增殖、周期及凋亡的影响并探究其作用机制。

方法

采用噻唑蓝（MTT）比色法检测三氧化二砷、双氢青蒿素、三氧化二砷联合双氢青蒿素对THP-1细胞增殖的影响，流式细胞术AnnexinV/碘化丙啶（PI）双染实验检测细胞周期和凋亡的变化，蛋白免疫印迹法（Western blot）检测细胞中半胱氨酸蛋白酶-3（Caspase-3），裂解半胱氨酸蛋白酶-3（cleaved Caspase-3），B淋巴细胞瘤-2（Bcl-2）和Bcl-2相关X蛋白（Bax）的表达，在高内涵显微镜下观察药物作用后细胞形态的变化。

结果

与空白组比较，不同浓度的三氧化二砷、双氢青蒿素对THP-1细胞的增殖均具有显著抑制作用（

0.01），且成剂量和时间依懒性；作用48 h，与同等剂量的三氧化二砷或双氢青蒿素单独应用比较，1 µmol·L

-1

三氧化二砷与2 µmol·L

-1

双氢青蒿素联合应用对THP-1细胞增殖的抑制作用显著增强（

0.01），且细胞阻滞在G

期（

0.01），并显著下调Bcl-2和Caspase-3的表达（

0.01），同时明显上调cleaved Caspase-3的表达水平（

0.05）。

结论

三氧化二砷联合双氢青蒿素对THP-1细胞具有明显抑制增殖和诱导凋亡的作用，其作用机制可能与阻滞细胞G

期，下调Bcl-2和Caspase-3蛋白表达，上调cleaved Caspase-3蛋白表达有关。

Abstract

Objective

To investigate the effects of arsenic trioxide combined with dihydroartemisinin on proliferation

cell cycle

and apoptosis of THP-1 cells

and explore the mechanism.

Method

The thiazolyl blue （MTT） method was applied to detect the effect of different concentrations of arsenic trioxide

dihydroartemisinin and arsenic trioxide combined with dihydroartemisinin on the proliferation of THP-1 cells. Annexin V/propidium iodide（PI）assay was used to detect the change of THP-1 cell cycle and apoptosis．Western blot was performed to assess the expression of cysteine protease-3（Caspase-3）

cleaved Caspase-3

B-lymphocytoma-2（Bcl-2） and Bcl-2 associated X protein （Bax）. The changes of cell morphology were observed under high intension microscope.

Result

Compared with blank group

arsenic trioxide and dihydroartemisinin both exhibited obvious antiproliferative effect on the human acute monocytic leukemia cell line THP-1 in time-dose dependence （

0.01）. After 48 h

compared with the same dose of arsenic trioxide or that of dihydroartemisinin alone

the inhibition effect of 1 µmol·L

-1

arsenic trioxide combined with 2 µmol·L

-1

dihydroartemisinin on proliferation of THP-1 cells was significantly stronger （

0.01）. Compared with the control group

arsenic trioxide combined with dihydroartemisinin significantly arrested the cells in G

phase （

0.01）

induced the downregulation of Caspase-3 and Bcl-2 （

0.01） and upregulation of cleaved Caspase-3 significantly（

0.05）.

Conclusion

Arsenic trioxide combined with dihydroartemisinin can significantly inhibit the proliferation and induce apoptosis of THP-1 cells. The possible mechanism may be related to arrest the cells in G

phase

reduce the expression of Caspase-3 and Bcl-2

increase the expression of cleaved Caspase-3.

关键词

Keywords

references

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