Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H<sub>2</sub>O<sub>2</sub>

Jing GUO; You-sheng MO; Qi WANG; Shui-qing HUANG

doi:10.13422/j.cnki.syfjx.20202038

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Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H₂O₂

Classic Prescriptions | 更新时间：2020-09-16

- Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H₂O₂
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 20, Pages: 39-45(2020)
- 作者机构：
  
  广州中医药大学科技创新中心，广州 510405
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20202038
  CLC： R2-0;R289;R54;R743
- Received：08 April 2020，
  
  Published Online：13 August 2020，
  
  Published：20 October 2020
- 稿件说明：
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郭静,莫友胜,王奇等.当归补血汤对内皮祖细胞氧化应激损伤的保护作用[J].中国实验方剂学杂志,2020,26(20):39-45.

GUO Jing,MO You-sheng,WANG Qi,et al.Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H₂O₂[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):39-45.
郭静,莫友胜,王奇等.当归补血汤对内皮祖细胞氧化应激损伤的保护作用[J].中国实验方剂学杂志,2020,26(20):39-45. DOI： 10.13422/j.cnki.syfjx.20202038.

GUO Jing,MO You-sheng,WANG Qi,et al.Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H₂O₂[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):39-45. DOI： 10.13422/j.cnki.syfjx.20202038.

摘要

目的

研究当归补血汤（DBT）对过氧化氢（H

）诱导的小鼠骨髓来源内皮祖细胞（EPCs）氧化应激损伤的保护作用。

方法

采用密度梯度离心法获取小鼠骨髓来源的单核细胞，通过特定培养基培养并鉴定获得EPCs，实验分为空白组，模型组，DBT组（100，200，400 mg·L

-1

）。采用噻唑蓝（MTT）比色法测定EPCs细胞存活率，构建H

诱导的细胞损伤模型；通过MTT，迁移小室（transwell），Matrigel基质胶，超氧化物荧光阴离子探针（DHE），分别检测EPCs增殖、迁移与体外血管形成能力，以及活性氧（ROS）水平，蛋白免疫印迹法（Western blot）检测细胞自噬相关蛋白泛素结合蛋白SQSTM1（p62），微管相关蛋白1轻链3Ⅱ型蛋白（LC3-Ⅱ）的蛋白表达。

结果

与空白组比较，模型组EPCs细胞增殖能力、迁移能力、成管腔数及小管分支长度均显著降低（

0.01），胞内ROS水平显著升高（

0.01），自噬标志性蛋白p62显著升高(

0.01），LC3-Ⅱ蛋白表达降低（

0.01）；与模型组比较，DBT组能呈浓度依赖性地升高EPCs增殖能力与迁移能力（

0.01），增加细胞成管腔数和小管分支长度（

0.01），降低胞内ROS水平（

0.01）；此外，DBT呈浓度依赖性上调了LC3-Ⅱ蛋白的表达（

0.01），降低p62蛋白的表达（

0.01）。

结论

当归补血汤能提高氧化应激状态下的EPCs自噬水平，促进损伤的EPCs迁移、增殖能力与血管形成能力，保护氧化应激条件下内皮祖细胞的生物学功能。

Abstract

Objective

To observe protective effect of Danggui Buxuetang (DBT) on oxidative stress injury of mouse bone marrow-derived endothelial progenitor cells (EPCs) against induced by hydrogen peroxide (H

Method

Monocytes from bone marrow of mice were obtained by density gradient centrifugation

and EPCs were obtained by specific culture medium. The experiment was divided into blank group，model group，DBT group (100，200，400 mg·L

-1

). Methyl thiazolyl tetrazolium（MTT） assay was used to determine the survival rate of EPCs and establish the cell injury model induced by H

. MTT，transwell chamber，matrigel and superoxide fluorescent anion probe (DHE) were used to detect the proliferation，migration，

in vitro

angiogenesis and ROS level，detection of autophagy by Western blot.

Result

Compared with blank group，the proliferation ability，migration ability，the number of lumens and the length of tubule branches of EPCs in the model group were significantly reduced (

0.01)，the level of intracellular reactive oxygen species （ROS） was significantly increased (

0.01)，the expression of p62，the light chain microtubule associated protein 1 protein light chain 3 Ⅱ type （LC3-Ⅱ） protein of microtubule associated protein 1，was significantly increased (

0.01). Compared with model group，DBT group increased the ability of cell proliferation and migration （

0.01）. In addition，DBT increased the expression of LC3-Ⅱ protein in a concentration dependent manner (

0.01)，and decreased the expression of p62 protein (

0.01).

Conclusion

DBT can improve the autophagy level of EPCs under oxidative stress

promote the proliferation

migration and angiogenesis of injured EPCs

and protect the biological function of EPCs under oxidative stress.

关键词

Keywords

references

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Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H2O2

DOI：10.13422/j.cnki.syfjx.20202038

摘要

Abstract

关键词

Keywords

references

Protective Effect of Danggui Buxuetang on Endothelial Progenitor Cells Against Oxidative Stress Injury Induced by H₂O₂