Effect of Bergapten on Apoptosis of HepG2 and Hep3B Cells Through PI3K/Akt Pathway

ZHAO Li-ping; Chao WANG; Nan TIAN; Chun-lei FAN

doi:10.13422/j.cnki.syfjx.20200623

您当前的位置：

首页 >

文章列表页 >

Effect of Bergapten on Apoptosis of HepG2 and Hep3B Cells Through PI3K/Akt Pathway

Pharmacology | 更新时间：2021-02-09

- Effect of Bergapten on Apoptosis of HepG2 and Hep3B Cells Through PI3K/Akt Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 6, Pages: 73-78(2020)
- 作者机构：
  
  浙江中医药大学　生命科学学院，杭州　 310053
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200623
  CLC： R22; R242; R2-031;R285.5
- Received：02 December 2019，
  
  Published Online：13 December 2019，
  
  Published：20 March 2020
- 稿件说明：
移动端阅览
ZHAO Li-ping , Chao WANG, Nan TIAN, et al. Effect of Bergapten on Apoptosis of HepG2 and Hep3B Cells Through PI3K/Akt Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(6): 73-78.
DOI：

ZHAO Li-ping , Chao WANG, Nan TIAN, et al. Effect of Bergapten on Apoptosis of HepG2 and Hep3B Cells Through PI3K/Akt Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(6): 73-78. DOI： 10.13422/j.cnki.syfjx.20200623.

摘要

目的：

探究佛手柑内酯通过磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路诱导人肝癌细胞HepG2和Hep3B凋亡的机制。

方法：

设立佛手柑内酯(5，50，200 μmol·L

-1

)组和空白组；采用噻唑蓝(MTT)比色法检测佛手柑内酯作用HepG2和Hep3B细胞24，48 h细胞增殖；磷脂结合蛋白V/碘化丙啶(Annexin V-FITC/PI)双染检测细胞凋亡；实时荧光定量逆转录聚合酶链反应(Real-time PCR)及蛋白免疫印迹法(Western bolt)检测相关mRNA和蛋白的表达含量；进行平板细胞克隆形成实验评价各组HepG2和Hep3B细胞的克隆形成率与效果。

结果：

MTT实验表明，佛手柑内酯能明显抑制HepG2和Hep3B细胞的增殖活性，且呈浓度依赖性；流式细胞仪分析结果表明，佛手柑内酯200 μmol·L

-1

组作用于HepG2和Hep3B细胞48 h，有明显促凋亡作用(

0.05)；Western blot结果显示，佛手柑内酯可以上调半胱氨酸蛋白酶(Caspase)-3，Caspase-8蛋白表达量(

0.05)，并且下调B淋巴细胞瘤-2(Bcl-2)，PI3K蛋白表达(

0.05)；Real-time PCR显示PI3K，Akt mRNA相对表达量降低(

0.05)；平板细胞克隆形成实验的结果显示，随着佛手柑内酯浓度的增加，各组细胞克隆形成率呈递减趋势，且佛手柑内酯200 μmol·L

-1

组作用降低明显(

0.05)。

结论：

佛手柑内酯对人肝癌细胞HepG2和Hep3B的增殖存在抑制作用，其可能是通过PI3K/Akt信号通路诱导HepG2和Hep3B细胞的凋亡。

Abstract

Objective:

To investigate the effect of bergapten on the apoptosis of HepG2 and Hep3B cells through phosphatidylinositol 3 kinase (PI3K)/protein kinase B(Akt) pathway.

Method:

Bergamot (5

200 μmol·L

-1

) groups and blank group were set up. The effect of bergapten at different concentrations on proliferation of HepG2 and Hep3B cells for 24

48 h were detected by thiazolyl blue(MTT) assay. Apoptosis was detected by Annexin V-FITC/propidium iodide double staining. Quantitative real-time fluorescence reverse transcriptional polymerase chain reaction (Real-time PCR) and Western blot assay were used to detect relevant mRNA and proteins expressions. The clone formation rate and the effect of HepG2 and Hep3B cells in each group were evaluated by plate cell clone formation.

Result:

MTT assay showed that bergapten could significantly inhibit the proliferation activity of HepG2 and Hep3B cells in a time-dependent manner. Flow cytometry analysis showed that bergapten in 200 μmol·L

-1

concentration groups had significant pro-apoptotic effect on HepG2 and Hep3B cells after 48 h (

0.05). Western blot results showed that bergamolactone could up-regulate the protein expressions of Caspase-3

Caspase-8 (

0.05)

and down-regulate protein expressions of B-lymphocytoma-2 (Bcl-2)

PI3K (

0.05). Real-time PCR results showed that mRNA expressions of PI3K and Akt were decreased(

0.05). The results of plate cell clone formation experiment showed that with the increase of the concentration of bergamolide

the cell clone formation rate of each group showed a decreasing trend

particularly in 200 μmol·L

-1

concentration group (

0.05).

Conclusion:

Bergapten can inhibit the proliferation of HepG2 and Hep3B cells

which may be induced through the PI3K/Akt signaling pathway.

关键词

Keywords

references

赵文昊，刘雪莲，李宏伟，等．原发性肝癌的营养代谢特点及意义 [J]．临床肝胆病杂志， 2019 ， 35 ( 4 ): 905 - 907 .

艾望，夏玉坤，王秀萍，等．原发性肝癌中医药治疗研究进展 [J]．辽宁中医药大学学报， 2018 ， 20 ( 3 ): 133 - 136 .

李红波，杨勇．孙思邈治疗肝脏相关病症方药运用特色 [J]．中医学报， 2018 ， 33 ( 1 ): 88 - 90 .

吴孝雄，吴申，陈挺松．肝脏癌邪及其临床应用 [J]．中医临床研究， 2017 ， 9 ( 12 ): 118 - 121 .

耿丹丹，赵博，王建华．佛手柑内酯对大鼠成骨细胞增殖与分化的影响 [J]．天然产物研究与开发， 2017 ， 29 ( 9 ): 1563 - 1567 .

贾恺宁，朱英．从肾论治原发性肝癌 [J]．时珍国医国药， 2017 ， 28 ( 4 ): 932 - 934 .

丁燕，傅友，单兰兰，等．佛手柑内酯对双氧水诱导人脐静脉内皮细胞衰老的影响 [J]．中国组织工程研究， 2016 ， 20 ( 46 ): 6885 - 6892 .

黄雪涛，陈泰华，祁敏倪，等．佛手柑内酯抑制肺癌细胞分泌可溶性白细胞介素2受体 [J]．中华临床医师杂志：电子版， 2016 ， 10 ( 11 ): 1567 - 1573 .

林碧华，马晓娟，万树伟，等．佛手柑内酯对鼻咽癌细胞凋亡的影响 [J]．肿瘤防治研究， 2014 ， 41 ( 11 ): 1163 - 1170 .

林碧华，万树伟，刘付梅，等．佛手柑内酯对鼻咽癌细胞周期的影响 [J]．中国药学杂志， 2014 ， 49 ( 10 ): 837 - 842 .

董芳，刘汉柱，孙阳，等．北沙参中佛手柑内酯的分离鉴定及体外抗肿瘤活性的初步测定 [J]．植物资源与环境学报， 2010 ， 19 ( 1 ): 95 - 96 .

刘大江，杨媛，刘畅，等．双酚A激活PI3K/Akt通路促进子宫内膜癌细胞增殖的研究 [J]. 国际妇产科学杂志， 2019 ， 46 ( 3 ): 259 - 262 .

吴林斌，吴元肇，李晓丹，等．连翘苷经PI3K/Akt信号通路干预肾细胞癌的机制研究 [J]．中草药， 2019 ， 50 ( 10 ): 2377 - 2382 .

刚小青，吴衡慧，周文磊．白桦脂醇通过抑制PI3K/Akt途径抑制宫颈癌小鼠移植瘤增殖 [J]．中国免疫学杂志， 2019 ， 35 ( 10 ): 1227 - 1231 .

周珍，刘少平，周朗，等．黄芩素通过PI3K/Akt通路增强三氧化二砷对肝癌细胞的促凋亡作用 [J]．现代肿瘤医学， 2019 ， 27 ( 10 ): 1661 - 1668 .

陈奕涵，张燕堂． PI3K/Akt信号通路在中药治疗肝癌过程中的研究进展 [J]．临床医药文献电子杂志， 2018 ， 5 ( 7 ): 179 - 182 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Sodium Cantharidinate and Vitamin B6 Injection Promotes Apoptosis of Hepatocellular Carcinoma Cell Line HepG2 through Regulating PI3K/Akt Signaling Pathway

Discussion on Effects and Mechanism of Ginseng Polysaccharide Injection on Proliferation Inhibition of Hepatoma Cells

Treatment of Sepsis-induced Inflammatory Responses with Xijiao Dihuangtang by Modulation of PKM2-mediated One-carbon Metabolism Pathway

Mitochondrial Quality Control Regulating Pathogenesis of Sarcopenia and Its Intervention by Traditional Chinese Medicine： A Review

Regulation of Signaling Pathways Related to Myocardial Infarction by Traditional Chinese Medicine： A Review

Related Author

ZHANG Meng

LI Sheng-chao

QIAO Zhi-bin

HE Hong-tao

PENG Li

GAO Heng-yu

GUO Lin-na

LIU Dan

Related Institution

Affiliated Hospital of Nanjing University of Chinese Medicine

The First Clinical Medical College，Nanjing University of Chinese Medicine

Jiangsu Province Hospital of Chinese Medicine

The First Affiliated Hospital of Henan University of Chinese Medicine

The First Clinical Medical School of Henan University of Chinese Medicine

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰