Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells

Zheng WEI; Ai-hua ZHANG; Ya-feng LI; Rui-feng LIANG; Wei-feng CUI; Yan-chun WANG; Jun-ping ZHANG

doi:10.13422/j.cnki.syfjx.20201927

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Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells

Pharmacology | 更新时间：2020-12-10

- Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 21, Pages: 62-67(2020)
- 作者机构：
  
  1.河南省中医药研究院，郑州 450004
  2.河南省人民医院，郑州 450000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20201927
  CLC： R22;R242;R2-031;R285.5
- Received：05 November 2019，
  
  Published Online：23 July 2020，
  
  Published：05 November 2020
- 稿件说明：
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魏征,张爱华,李亚峰等.化瘀解毒方对胃癌细胞凋亡的作用及机制[J].中国实验方剂学杂志,2020,26(21):62-67.

WEI Zheng,ZHANG Ai-hua,LI Ya-feng,et al.Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):62-67.
魏征,张爱华,李亚峰等.化瘀解毒方对胃癌细胞凋亡的作用及机制[J].中国实验方剂学杂志,2020,26(21):62-67. DOI： 10.13422/j.cnki.syfjx.20201927.

WEI Zheng,ZHANG Ai-hua,LI Ya-feng,et al.Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):62-67. DOI： 10.13422/j.cnki.syfjx.20201927.

摘要

目的

观察化瘀解毒方（Huayu Jiedu recipe，HYJDR）对胃癌细胞凋亡的影响，探讨HYJDR对胃癌细胞凋亡的作用机制，为HYJDR的临床运用提供实验依据。

方法

运用噻唑蓝（thiazolyl blue tetrazolium bromide，MTT）比色法检测HYJDR（5，10，15，20，25，30 g·L

-1

）对胃癌SGC-7901细胞活力影响，测定HYJDR对SGC-7901细胞的半数抑制浓度（median inhibition concentration，IC

）。利用碘化丙啶（propidium iodide，PI）和hoechst双荧光染色检测HYJDR对胃癌SGC-7901细胞的凋亡影响情况。蛋白免疫印迹法（Western blot）检测HYJDR对胃癌凋亡相关蛋白表达情况的影响。实时荧光聚合酶链反应（Real-time-polymerase chain reaction，Real-time PCR）检测HYJDR对胃癌细胞凋亡相关蛋白信使RNA（mRNA）表达水平的影响。

结果

与空白组比较，HYJDR质量浓度

10 g·L

-1

时可以明显抑制胃癌细胞的增殖活力，随着HYJDR浓度的增加，细胞活力下降明显（

0.05），呈明显的浓度依赖性。HYJDR可以促进胃癌细胞的凋亡，随着HYJDR浓度的增加，细胞的凋亡逐渐增多。与空白组比较，HYJDR可以明显抑制胃癌细胞凋亡相关蛋白B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）表达（

0.05），同时提高Bcl-2细胞凋亡抵抗蛋白（Bcl-2 antagonist of cell death， Bad）和Bcl-2相关蛋白X（Bcl-2-associated X， Bax）等促凋亡蛋白表达（

0.05）。PCR结果显示，与空白组比较，HYJDR（5，10 g·L

-1

）组可以有效抑制胃癌细胞中Bcl-2 mRNA表达（

0.05），HYJDR各组促进Bad mRNA表达（

0.05）。

结论

HYJDR可以明显促进胃癌细胞SGC-7901的凋亡，其作用很可能是通过提高Bad mRNA表达，抑制Bcl-2蛋白表达来实现的。

Abstract

Objective

To observe the effect of Huayu Jiedu recipe （HYJDR） on apoptosis of gastric cancer cells， explore the mechanism of HYJDR on apoptosis of gastric cancer cells and provide experimental basis for clinical application of HYJDR.

Method

Thiazolyl blue tetrazolium bromide （MTT） method was used to detect the effect of HYJDR （5，10，15，20，25，30 g·L

-1

） on the activity of SGC-7901 cells and the median inhibition concentration （IC

） of HYJDR on SGC-7901 cells. Propidium iodide （PI） and Hoechst double fluorescence staining were used to detect the effect of HYJDR on the apoptosis of gastric cancer SGC-7901 cells. Western blot was used to detect the effect of HYJDR on the expression of apoptosis related proteins in gastric cancer cells. Real-time polymerase chain reaction （Real-time PCR） was used to detect the effect of HYJDR on the expression of apoptosis related protein mRNA in gastric cancer cells.

Result

As compared with blank group， HYJDR （

10 g·L

-1

） can significantly inhibit the proliferation of gastric cancer cells. With the increase of the concentration of HYJDR， the cell viability decreased significantly obviously in a concentration-dependent manner（

0.05）. HYJDR can significantly promote the apoptosis of gastric cancer cells. With the increase of the concentration of HYJDR， the apoptosis of gastric cancer cells gradually increased. As compared with blank group， HYJDR can obviously inhibit the expression of apoptosis-related B -cell lymphoma-2 （Bcl-2）（

0.05）， and increase the expression of apoptosis-promoting proteins such as Bcl-2 antagonist of cell death （Bad） and Bcl-2-associated X （Bax）（

0.05）. Real-time PCR results showed that as compared with blank group， HYJDR（5，10 g·L

-1

） could effectively inhibit the expression of Bcl-2 protein mRNA in gastric cancer cells（

0.05）， and all HYJDR groups could promote the expression level of Bad mRNA（

0.05）.

Conclusions

HYJDR can obviously promote the apoptosis of SGC-7901， and its effect is probably achieved by increasing the expression of Bad mRNA and inhibiting the expression of Bcl-2 protein.

关键词

Keywords

references

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