Regulatory effect of Gpr124 on proliferation, migration and angiogenesis of retinal microvascular endothelial cells under high-glucose condition

Yuwen WANG; Furong LI; Rongdi YUAN

doi:10.16016/j.2097-0927.202403058

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Regulatory effect of Gpr124 on proliferation, migration and angiogenesis of retinal microvascular endothelial cells under high-glucose condition

Basic Medicine | 更新时间：2026-01-09

- Regulatory effect of Gpr124 on proliferation, migration and angiogenesis of retinal microvascular endothelial cells under high-glucose condition
- Journal of Army Medical University Vol. 46, Issue 18, Pages: 2101-2109(2024)
- 作者机构：
  
  400037 重庆，陆军军医大学(第三军医大学)第二附属医院眼科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202403058
  CLC： R322.13;R363.13;R774.1
- Received：13 March 2024，
  
  Revised：2024-04-30，
  
  Published：30 September 2024
- 稿件说明：
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Yuwen WANG, Furong LI, Rongdi YUAN. Regulatory effect of Gpr124 on proliferation, migration and angiogenesis of retinal microvascular endothelial cells under high-glucose condition[J]. Journal of Army Medical University, 2024, 46(18): 2101-2109.
DOI：

Yuwen WANG, Furong LI, Rongdi YUAN. Regulatory effect of Gpr124 on proliferation, migration and angiogenesis of retinal microvascular endothelial cells under high-glucose condition[J]. Journal of Army Medical University, 2024, 46(18): 2101-2109. DOI： 10.16016/j.2097-0927.202403058.

摘要

目的

研究高糖环境下人视网膜微血管内皮细胞(human retinal microvascular endothelial cell，HRMEC)中G蛋白偶联受体124(Gpr124)表达的变化，以及干扰Gpr124对HRMEC的调控作用。

方法

免疫荧光观察Gpr124在HRMEC中的表达；高糖刺激HRMEC，将细胞分为空白组和高糖组，CCK-8、EdU检测细胞活力与增殖情况；病毒抑制Gpr124表达后，将细胞分为空白对照组、高糖对照组、高糖+shRNA组以及高糖+Gpr124 shRNA组，划痕实验评估细胞迁移，基质凝胶评估细胞小管形成能力。Western blot检测Gpr124、VEGFA、MMP9和β-catenin相对蛋白表达水平。

结果

相较于空白组，HRMEC在高糖组中显示出显著提升的细胞活力和增殖细胞数(

0.01)。高糖环境下，Gpr124、VEGFA以及MMP9的表达水平也显著增加(

0.01)。同时，在各个时间点，高糖对照组的迁移面积以及体外血管形成面积和管径长度均显著高于空白对照组(

0.01)。然而，在敲低Gpr124的表达后，高糖+Gpr124 shRNA组的HRMEC在迁移和体外血管形成能力上相较于高糖+shRNA组有显著的降低(

0.01)，并且高糖+Gpr124 shRNA组相比于高糖+shRNA组VEGFA、MMP9以及β-catenin蛋白表达减少(

0.01)。

结论

Gpr124能够调控高糖环境下HRMEC的增殖、迁移以及管形成能力，并且还可以抑制VEGFA和MMP9的过度表达，这一影响可能是通过调节Wnt/β-catenin经典通路来实现的。

Abstract

Objective

To investigate the changes in G protein-coupled receptor 124 (Gpr124) expression in human retinal microvascular endothelial cells (HRMEC) under high-glucose condition and the regulatory effect of Gpr124 interference on HRMEC.

Methods

Immunofluorescence assay was used to observe the expression of Gpr124 in HRMEC. The cells were divided into a blank group and a high glucose group. CCK-8 assay and EdU cell proliferation assay were used to detect cell viability and proliferation

respectively. The Gpr124 expression was knocked down by transducting lentiviral virus carrying shRNA targeting Gpr124. Thus

the cells were divided into a blank control group

a high glucose control group

a high glucose+shRNA group

and a high glucose+Gpr124 shRNA group. Cell scratch test was performed to evaluate cell migration

and Matrigel plug assay was employed to assess the cell tubule formation. Western blotting was applied to detect the protein levels of Gpr124

VEGFA

MMP9

and β-catenin.

Results

The HRMEC from the high glucose group showed enhanced cell viability and increased number of proliferating cells compared to the cells of the blank group (

0.01). High glucose treatment also induced increased expression levels of Gpr124

VEGFA and MMP9 (

0.01)

and larger migratory area and

in vitro

angiogenic area as well as longer tube diameter at all time points when compared with the conditions in the blank control group (

0.01). However

knockdown of Gpr124 resulted in a significant decrease in HRMEC migration and

in vitro

angiogenic capacity

accompanied by decreases in protein expression of VEGFA

MMP9 and β-catenin (

0.01).

Conclusion

Gpr124 can regulate the proliferation

migration

and tube formation ability of HRMEC under high-glucose condition

and also inhibit the overexpression of VEGFA and MMP9

which may be through regulating the classical Wnt/β-catenin pathway.

关键词

Keywords

references

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