六神丸和雄黄诱导子宫内膜癌细胞凋亡和DNA损伤的机制

刘正芸; 勾英; 姜念; 王欢; 刘杰

doi:10.13422/j.cnki.syfjx.20201421

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六神丸和雄黄诱导子宫内膜癌细胞凋亡和DNA损伤的机制

药理 | 更新时间：2020-09-09

- 六神丸和雄黄诱导子宫内膜癌细胞凋亡和DNA损伤的机制
- Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells
- 中国实验方剂学杂志 2020年26卷第14期页码：99-104
- 作者机构：
  
  遵义医科大学，贵州遵义 563000
- 作者简介：
  
  刘正芸，硕士，助理实验师，从事抗肿瘤相关研究，E-mail：916103898@qq.com
  刘杰，博士，研究员，从事药理和药物代谢研究，E-mail：Jieliu@zmu.edu.cn
- 基金信息：
  
  贵州省普通高等学校特色药物肿瘤防治特色重点实验室开放基金项目(TSYW-2016003);遵义医学院硕士科研启动基金项目(F-902)
- DOI：10.13422/j.cnki.syfjx.20201421
  中图分类号： R22;R242;R2-031;R285.5
- 纸质出版日期：2020-07-20，
  
  网络出版日期：2020-05-06，
- 稿件说明：
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刘正芸,勾英,姜念等.六神丸和雄黄诱导子宫内膜癌细胞凋亡和DNA损伤的机制[J].中国实验方剂学杂志,2020,26(14):99-104.

LIU Zheng-yun,GOU Ying,JIANG Nian,et al.Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(14):99-104.
刘正芸,勾英,姜念等.六神丸和雄黄诱导子宫内膜癌细胞凋亡和DNA损伤的机制[J].中国实验方剂学杂志,2020,26(14):99-104. DOI： 10.13422/j.cnki.syfjx.20201421.

LIU Zheng-yun,GOU Ying,JIANG Nian,et al.Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(14):99-104. DOI： 10.13422/j.cnki.syfjx.20201421.

摘要

目的

比较经典名药六神丸和雄黄（As

）对子宫内膜癌细胞JEC的抗肿瘤作用及其机制。

方法

原子吸收法测定六神丸与As

中砷（As）的溶解度；噻唑蓝（MTT）比色法比较六神丸与As

对细胞增殖的抑制作用，细胞迁移实验比较六神丸与As

对细胞迁移的抑制作用；磷脂酰丝氨酸蛋白抗体/碘化丙啶（Annexin V-FITC/PI）双染结合流式细胞术比较六神丸与As

对细胞凋亡的影响，蛋白免疫印迹法（Western blot）比较六神丸与As

对凋亡和DNA损伤相关蛋白的影响。

结果

原子吸收法测定结果显示六神丸的As溶出量为17.4%，As

的As溶出量仅为1.6%。但在相同As含量的情况下，与As

组比较，六神丸3，10 mg·L

-1

组的细胞存活率明显降低（

0.05），六神丸0.25，0.5，1 mg·L

-1

组早期凋亡率明显增高（

0.05），六神丸1 mg·L

-1

组细胞迁移率明显下降（

0.05），六神丸1 mg·L

-1

组剪切的半胱氨酸天冬氨酸蛋白酶-3（cleaved Caspase-3），剪切的半胱氨酸天冬氨酸蛋白酶-7（cleaved Caspase-7），剪切的聚ADP核糖聚合酶（cleaved PARP）以及磷酸化的组蛋白（p-H

AX），磷酸化的检测点激酶2（p-CHK2），人共济失调毛细血管扩张症突变激酶（ATM）蛋白表达水平升高（

0.05），人共济失调毛细血管扩张症Rad3相关激酶（ATR）蛋白表达水平降低（

0.05），而Caspase-3和Caspase-7蛋白表达水平无明显改变。

结论

在等As剂量下，六神丸与As

均能抑制JEC细胞的增殖和迁移，诱导细胞凋亡和DNA损伤，但六神丸的效果优于As

，提示中药复方六神丸中还有其他成分协同As共同参与抗肿瘤作用。

Abstract

Objective

Compare the anti-tumor effect and mechanism of Liushenwan and realgar (As

) on human endometrial cancer cells JEC.

Method

The release of As

in Liushenwan and As

was measured by atomic absorption spectrometry. The 3-(4

5-dimethylthiazol-2-yl)-2

5-diphenyltetrazolium bromide (MTT) assay was used for cell proliferation

and cell migration was measured by Transwell assay. Flow cytometry and Western Blot were used to determine apoptosis and DNA damage.

Result

The dissolution of As in Liushenwan was 17.4%

and that of As

was only 1.6% according to atomic absorption assay. With the same content of As

compared with the As

group

the cell viability in the 3，10 mg·L

-1

Liushenwan groups was decreased (

0.05)

the early apoptosis rate was significantly increased in 0.25，0.5，1 mg·L

-1

Liushenwan groups (

0.05)

the rate of cell migration was decreased in 1 mg·L

-1

Liushenwan group (

0.05)

the expressions of cleaved cysteinyl aspartate-specific proteinase-3 (cleaved Caspase-3)

cleaved cysteinyl aspartate-specific proteinase-7 (cleaved Caspase-7)

cleaved poly ADP-ribose polymerase (cleaved PARP)

phosphorylated histone (p-H

AX)

phosphorylation of checkpoint kinase 2 (p-CHK2) and ataxia-telangiectasia mutated (ATM) were increased in 1 mg·L

-1

Liushenwan group (

0.05)

while the expression of phosphorylation of ataxia-telangiectasia mutated rad3-related (ATR) was decreased in 1 mg·L

-1

Liushenwan group (

0.05)

with no significant changes in the expressions of cysteinyl aspartate-specific proteinase-3 (Caspase-3) and cysteinyl aspartate-specific proteinase-7 (Caspase-7).

Conclusion

With the same content of As

both Liushenwan and As

could inhibit JEC cell proliferation and migration

and induce cell apoptosis and DNA damage. Liushenwan has a stronger effect than As

. It is suggested that there are other components in Liushenwan with an anti-tumor effect in cooperation with As.

关键词

六神丸雄黄砷子宫内膜癌细胞凋亡DNA损伤

Keywords

Liushenwanrealgar (As4S4)arsenic (As)endometrial cancerapoptosisDNA damage

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