GATA2/FGF21 Axis Regulates the Effects of High Glucose on the Apoptosis, Autophagy and Oxidative Stress of Human Umbilical Vein Endothelial Cell via PI3K/AKT/mTOR Pathway

Ann Clin Lab Sci. 2022 Mar;52(2):278-291.

Abstract

Objective: We investigated the role and mechanism of GATA-binding factor 2/Fibroblast growth factor 21 (GATA2/FGF21) axis in high glucose (HG)-induced injury in human umbilical vein endothelial cells (HUVEC).

Methods: After HG treatment and transfection, the viability and apoptosis of HUVECs were determined via Cell Counting Kit-8 and Hoechst 33258 staining methods, and the content of lactate dehydrogenase (LDH) and reactive oxygen species (ROS) was measured via colorimetric assay and DCFH-DA staining. The potential transcription factor of FGF21 was predicted with bioinformatic analysis and confirmed via dual-luciferase reporter assay and chromatin immunoprecipitation. The expressions of GATA2/FGF21, apoptosis-, autophagy- and phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway-related factors were quantified with quantitative real-time polymerase chain reaction or Western blot.

Results: Overexpressed FGF21 abolished the effects of HG on repressing the expressions of FGF21 and cell viability, and promoting apoptosis, the levels of LDH and ROS and autophagy in HUVECs, with increased Bcl-2 and p62 expression yet decreased Bax, Cleaved PARP, Cleaved caspase-3, LC3 II/LC3 I ratio and Beclin 1. GATA2 was the transcription factor of FGF21 and was downregulated after HG treatment, and the effects of overexpressed FGF21 in HG-treated HUVECs were all reversed after the silence of GATA2. Besides, overexpressed FGF21 promoted the activation of PI3K/AKT/mTOR pathway, with increased phosphorylation levels of PI3K, AKT and mTOR, whereas silencing GATA2 abolished the trend.

Conclusion: GATA2/FGF21 axis has a protective function against HG in HUVEC via regulating PI3K/AKT/mTOR pathway.

Keywords: GATA-binding factor 2/Fibroblast growth factor 21 (GATA2/FGF21) axis; Gestational diabetes mellitus; Human umbilical vein endothelial cells; phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) pathway.

MeSH terms

  • Apoptosis
  • Autophagy
  • Fibroblast Growth Factors
  • GATA2 Transcription Factor
  • Glucose / metabolism
  • Glucose / pharmacology
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Oxidative Stress
  • Phosphatidylinositol 3-Kinase* / metabolism
  • Phosphatidylinositol 3-Kinase* / pharmacology
  • Phosphatidylinositol 3-Kinases* / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Reactive Oxygen Species / metabolism
  • Signal Transduction
  • TOR Serine-Threonine Kinases / metabolism
  • Transcription Factors / metabolism

Substances

  • GATA2 Transcription Factor
  • GATA2 protein, human
  • Reactive Oxygen Species
  • Transcription Factors
  • fibroblast growth factor 21
  • Fibroblast Growth Factors
  • MTOR protein, human
  • Phosphatidylinositol 3-Kinase
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Glucose