Effect of metformin on autophagy in human umbilical vein endothelial cells

Bingyu LI; Weiyuan LIU; Xin LIN; Zhiying ZHAO; Zhi WANG; Yangdong LIU; Yun WANG

doi:10.16016/j.1000-5404.201911118

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Effect of metformin on autophagy in human umbilical vein endothelial cells

更新时间：2026-01-09

- Effect of metformin on autophagy in human umbilical vein endothelial cells
- Journal of Third Military Medical University Vol. 42, Issue 8, Pages: 783-789(2020)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the Talent Support Program of Army Medical University （Third Military Medical University）(2019);the Science and Technology Research Project of Chongqing Education Commission(KJ1600208);the Foundation of Health and Family Planning Commission of Chongqing(2016ZDXM004);the General Program of National Natural Science Foundation of China(81470963)
- DOI：10.16016/j.1000-5404.201911118
  CLC：
- Received：14 November 2019，
  
  Published：30 April 2020
- 稿件说明：
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Bingyu LI, Weiyuan LIU, Xin LIN, et al. Effect of metformin on autophagy in human umbilical vein endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(8): 783-789.
DOI：

Bingyu LI, Weiyuan LIU, Xin LIN, et al. Effect of metformin on autophagy in human umbilical vein endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(8): 783-789. DOI： 10.16016/j.1000-5404.201911118.

摘要

Abstract

Objective

To observe the effects of metformin on the autophagy of endothelial cells and explore the underlying mechanism.

Methods

Human umbilical vein endothelial cells (HUVECs) were treated with 0

2.5

20 and 40 mmol/L metformin for 24 h or with 5 mmol/L metformin for 24

48 and 72 h

and cell survival after the treatment was assessed using CCK-8 assay. The formation of autophagosomes in HUVECs treated with metformin and transforming growth factor-β1 (TGF-β1)

alone or in combination

was observed with transmission electron microscopy

and the expression levels of autophagy-related molecules LC3II and Beclin-1 were detected using immunofluorescence assay and Western blotting. The effects of XMU-MP-1 (an inhibitor of Hippo signaling pathway) and verteporfin (a blocker of Hippo signal transcription factor YAP) in the absence or presence of metformin or TGF-β1 on the expression levels of LC3II

YAP

pYAP and CTGF (the target protein of pYAP) were detected using Western blotting.

Results

CCK-8 assay showed that compared with the control cells

HUVECs treated with 10

20 and 40 mmol/L metformin for 24 h exhibited significantly reduced cell survival rates (all

0.01)

and we therefore used 5 mmol/L metformin for 24 h in the subsequent experiments. The results of electron microscopy

immunofluorescence assay and Western blotting showed that treatment with TGF-β1 significantly increased the number of autophagosomes

enhanced the

expression of LC3II and Beclin-1 (

0.05)

lowered the expression of pYAP (

0.01)

and up-regulated CTGF expression in HUVECs (

0.01). These changes were significantly blocked by metformin. The expression of levels LC3II and CTGF were also inhibited in the cells by treatment with verteporfin and TGF-β1 (

0.01). XMU-MP-1 showed similar effects to TGF-β1 and significantly up-regulated the expression of LC3II (

0.01)

decreased the expression of pYAP (

0.01)

and enhanced the expression of CTGF (

0.01); such effects of XMU-MP-1 were also significantly blocked by metformin (all

0.05). The expression levels of CTGF and LC3II were obviously down-regulated in the cells treated with both verteporfin and XMU-MP-1 (

0.01).

Conclusion

Metformin may protect the veins by inhibiting TGF-β1-induced autophagy and blocking the activation of YAP signaling in the endothelial cells.

关键词

Keywords

references

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