Effects of exosomes derived from placental mesenchymal stem cells on high glucose-induced senescence in human dermal fibroblasts

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Effects of exosomes derived from placental mesenchymal stem cells on high glucose-induced senescence in human dermal fibroblasts

Basic Medicine | 更新时间：2026-01-09

- Effects of exosomes derived from placental mesenchymal stem cells on high glucose-induced senescence in human dermal fibroblasts
- Journal of Third Military Medical University Vol. 42, Issue 12, Pages: 1163-1173(2020)
- 作者机构：
  
  1.300070 天津，天津医科大学研究生院
  2.100048 北京，中国人民解放军总医院创新医学研究部组织修复与再生研究中心，皮肤损伤修复与再生北京市重点实验室
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81571905);the National Key Research Development Plan(2017YFC1103300)
- DOI：10.16016/j.1000-5404.202001190
  CLC： R322.99; R329.28; R587.102
- Received：22 January 2020，
  
  Revised：2020-03-08，
  
  Published：30 June 2020
- 稿件说明：
移动端阅览
Xiaowei BIAN, Cuiping ZHANG, Bingmin LI, et al. Effects of exosomes derived from placental mesenchymal stem cells on high glucose-induced senescence in human dermal fibroblasts[J]. Journal of Third Military Medical University, 2020, 42(12): 1163-1173.
DOI：

Xiaowei BIAN, Cuiping ZHANG, Bingmin LI, et al. Effects of exosomes derived from placental mesenchymal stem cells on high glucose-induced senescence in human dermal fibroblasts[J]. Journal of Third Military Medical University, 2020, 42(12): 1163-1173. DOI： 10.16016/j.1000-5404.202001190.

摘要

目的

2

探索高糖(high glucose

HG)促进体外培养的人皮肤成纤维细胞(human dermal fibroblasts

HDFs)的衰老条件，建立高糖老化模型；观察胎盘间充质干细胞来源的外泌体(exosomes

Exos)对高糖培养的成纤维细胞增殖、迁移及衰老的影响。

方法

2

分离人包皮组织的HDFs。实验分为3组：正常对照组(5.5 mmol/L葡萄糖)、HG1组(26 mmol/L葡萄糖)、HG2组(35 mmol/L葡萄糖)。CCK-8试剂盒检测各组细胞在1、3、5、7 d的增殖情况；各组培养7 d后进行划痕实验，观察各组HDFs在24 h内的迁移率；2′，7′二氯荧光素二醋酸(DCFH)法检测各组培养7 d后细胞内活性氧簇(ROS)水平；细胞衰老β-半乳糖苷酶染色试剂盒检测各组培养7 d后β-半乳糖苷酶活性。体外培养胎盘间充质干细胞(placenta mesenchymal stem cells

PMSCs)，差速高速离心法分离Exos。将上述培养的HDFs分为3组：HG2组(35 mmol/L葡萄糖)、HG2+低浓度Exos(5 μg/mL)组、HG2+高浓度Exos(50 μg/mL)组，培养3 d后，检测细胞增殖情况、迁移率、ROS水平和β-半乳糖苷酶活性；流式细胞术检测细胞周期；Western blot分析衰老相关蛋白p53和p21表达情况。

结果

2

与对照组相比，HG1和HG2组培养的HDFs增殖、迁移能力明显下降(

P

<

0.01)，ROS水平显著升高(

P

<

0.01)，β-半乳糖苷酶活性显著增加(

P

<

0.01)，而且各指标尤其以HG2组变化最为明显；Exos实验结果显示：高浓度Exos明显促进高糖培养HDFs的增殖和迁移(

P

<

0.01)，显著降低细胞内的ROS水平和β-半乳糖苷酶活性(

P

<

0.01)，同时使细胞内p53和p21蛋白的表达水平显著下降(

P

<

0.01)，而低浓度Exos无明显效果。

结论

2

胎盘间充质干细胞来源的Exos促进高糖培养的HDFs增殖、迁移，缓解细胞的氧化应激损伤和衰老状态，其机制可能与衰老相关蛋白p53和p21表达下调有关。

Abstract

Objective

2

To explore the optimal conditions for inducing senescence in human dermal fibroblasts (HDFs) cultured in high glucose and observe the effect of exosomes derived from human placental mesenchymal stem cells (PMSCs) on the proliferation

migration and senescence of high glucose-induced fibroblasts.

Methods

2

HDFs were isolated from human foreskin tissue and cultured in the presence of 5.5 mmol/L glucose (normal control group)

26 mmol/L glucose (HG1 group) and 35 mmol/L glucose (HG2 group). CCK-8 assay was used to detect cell proliferation in each group on days 1

3

5 and 7 of cell culture. The cells cultured for 7 d were tested for migration within 24 h using cell scratch assay; reactive oxygen species (ROS) production in the cells was detected using 2′

7′-dichlorofluorescein diacetic acid (DCFH) method

and β-galactosidase activity was analyzed using a β-galactosidase staining kit. The HDFs cultured in 35 mmol/L glucose for 7 d were exposed for 3 days to 5 μg/mL and 50 μg/mL exosomes

which were isolated from human PMSCs by differential ultracentrifugation

and the cell proliferation

migration

ROS levels and β-galactoside activity were assessed. Flow cytometry was used to analyze the cell cycle changes and Western blotting was performed to detect the expression of aging-related proteins p53 and p21 in the cells following treatment with the exosomes.

Results

2

Compared with the control cells

the HDFs in HG1 and HG2 groups exhibited significantly decreased proliferation and migration ability (

P

<

0.01) and increased ROS production (

P

<

0.01) and β-galactoside activity (

P

<

0.01). Treatment with the exosomes obviously promoted the proliferation and migration of HDFs cultured in high glucose (

P

<

0.01). At the concentration of 50 μg/mL

PMSC-derived exosomes significantly decreased ROS production and β-galactoside activity (

P

<

0.01) and obviously down-regulated the expression levels of p53 and p21 (

P

<

0.01) in the HDFs cultured in high glucose

whereas a low concentration (5 μg/mL) of the exosomes did not produce such effects.

Conclusion

2

Exosomes derived from PMSCs promote the proliferation and migration and alleviate oxidative stress-induced damage and cell senescence in high glucose-induced HDFs

the mechanism of which may involve the down-regulation of the aging-related proteins p53 and p21.

关键词

Keywords

references

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