小檗碱对高糖环境下HCCLM3细胞凋亡与自噬的影响

刘倩; 傅缨; 资晓飞; 熊耀斌; 李媛

doi:10.13422/j.cnki.syfjx.20182322

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小檗碱对高糖环境下HCCLM3细胞凋亡与自噬的影响

药理 | 更新时间：2021-02-09

- 小檗碱对高糖环境下HCCLM3细胞凋亡与自噬的影响
- Effect of Berberine on HCCLM3 Cells Apoptosis and Autophagy Under High Glucose Condition
- 中国实验方剂学杂志 2019年25卷第2期页码：68-73
- 作者机构：
  
  1.南昌大学　第二附属医院，南昌　 330006；
  2.江西省医学科学院，南昌　 330006；
  3.南昌大学医院，南昌　 330006
- 作者简介：
  
  刘倩，硕士，从事中西医结合内科研究，E-mail：501170789@qq.com
  *傅缨，硕士，教授，硕士生导师，主任中医师，从事中医药肿瘤治疗与康复的研究，E-mail：yingfu-116@163.com
- 基金信息：
  
  江西省卫生计生委课题项目(A098;20185252)
- DOI：10.13422/j.cnki.syfjx.20182322
  中图分类号： R22; R242; R2-031; R273; R285.5
- 收稿日期：2018-04-27，
  
  网络出版日期：2018-09-12，
  
  纸质出版日期：2019-01-20
- 稿件说明：
移动端阅览
刘倩, 傅缨, 资晓飞, 等. 小檗碱对高糖环境下HCCLM3细胞凋亡与自噬的影响[J]. 中国实验方剂学杂志, 2019,25(2):68-73.

Qian LIU, Ying FU, Xiao-fei ZI, et al. Effect of Berberine on HCCLM3 Cells Apoptosis and Autophagy Under High Glucose Condition[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(2): 68-73.
刘倩, 傅缨, 资晓飞, 等. 小檗碱对高糖环境下HCCLM3细胞凋亡与自噬的影响[J]. 中国实验方剂学杂志, 2019,25(2):68-73. DOI： 10.13422/j.cnki.syfjx.20182322.

Qian LIU, Ying FU, Xiao-fei ZI, et al. Effect of Berberine on HCCLM3 Cells Apoptosis and Autophagy Under High Glucose Condition[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(2): 68-73. DOI： 10.13422/j.cnki.syfjx.20182322.

摘要

目的：

探讨小檗碱(berberine，BBR)对高糖环境下人肝癌HCCLM3细胞增殖、凋亡及自噬相关基因表达的影响，并探究其可能的抗肿瘤分子机制。

方法：

把HCCLM3细胞加入低、中、高质量浓度葡萄糖中，通过细胞增殖实验(cell counting kit-8，CCK-8)检测发现高质量浓度葡萄糖对HCCLM3细胞增殖的作用最明显。基于此，选择高糖培养HCCLM3细胞，加入BBR(5，10，20，30，40，50 μmol·L

－1

)进行体外干预24 h，研究各组药物对HCCLM3细胞增殖的抑制效果，并设顺铂组对照，通过流式细胞仪观察细胞凋亡率的变化，实时荧光定量PCR(Real-time PCR)检测B淋巴细胞瘤-2(B-cell lymphoma-2，Bcl-2)和自噬相关基因Atg5，Beclin1 mRNA表达水平。

结果：

与空白组比较，随着葡萄糖浓度的增加，HCCLM3细胞在高糖组中增殖能力最强(

0.05)；BBR对高糖培养下HCCLM3增殖的抑制作用与BBR浓度有相关性，BBR(5，10，20，30，40，50 μmol·L

－1

)组抑制率依次为33.86%，40.75%(

0.05)，49.22%，55.1%，60.12%，61.42%(

0.01)；细胞凋亡率随BBR浓度增加有显著上升(

0.01)；与单纯高糖组比较，各干预组中凋亡基因Bcl-2与自噬基因Atg5，Beclin1 mRNA表达上调(

0.05，

0.01)。

结论：

BBR对高糖环境下HCCLM3细胞增殖具有抑制作用，可能是通过调控Bcl-2的表达，诱导该细胞凋亡，同时又上调Beclin1，Atg5 mRNA表达水平，促进自噬发挥抗肿瘤作用。

Abstract

Objective:

To explore the anti-tumor molecular mechanism of berberine (BBR)by observing and analyzing its effect on proliferation

apoptosis and autophagy-related gene expression for HCCLM3 cells under high glucose condition.

Method:

HCCLM3 cells were added into low

medium or high-concentration groups of glucose. It was found in cell counting kit-8(CCK-8)that the high concentration of glucose had the most obvious effect on HCCLM3 cells proliferation. Based on the above experimental result

HCCLM3 cells treated with high concentration of glucose was selected and then different concentrations of berberine (5

50 μmol·L

-1

) was added for

in vitro

intervention for 24 h. Then the effect of each drug group on the proliferation of HCCLM3 cells were studied. At the same time

the control group of metformin was arranged. After that

the changes of apoptosis rate were observed by flow cytometry

and the expression levels of B-cell lymphoma-2(Bcl-2)and autophagy genes Atg5

Beclin1 were detced by Real-time polymerase chain reaction (Real-time PCR).

Result:

With the increase of glucose concentration

HCCLM3 cell had the strongest migration and proliferation ability in high glucose group (

0.05). The inhibitory effect of BBR on the proliferation of HCCLM3 in high glucose culture was correlated with the concentration of BBR(5

50 μmol·L

-1

)

and the inhibition rate was 33.86%

40.75% (

0.05)

49.22%

55.1%

60.12%

and 61.42% respectively from low concentration to high concentration (

0.01). The apoptosis rate was significantly increased with the increase of BBR concentration (

0.01). The expression levels of Bcl-2

Atg5 and Beclin1 gene were significantly up-regulated in all intervening groups as compared with those in single high glucose group (

0.05

0.01).

Conclusion:

BBR could inhibit the proliferation of HCCLM3 cells in high glucose environment. Its inhibition effect for HCCLM3 cells might be achieved by inducing apoptosis of the cells

regulating Bcl-2 and up-regulating the expression levels of autophagy gene Beclin1 and Atg5.Thus BBR plays an anti-tumor role through promoting autophagy in high glucose environment.

关键词

Keywords

references

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