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miR-16-5p Regulates Proliferation and Apoptosis in High Glucose-Treated Human Retinal Microvascular Endothelial Cells

JianFeng Zhao1, YanFei Zhang1, Yuan Xia1

  • 1Department of Ophthalmology, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan, China.

Journal of Ophthalmology
|April 1, 2025
PubMed

Insights

MicroRNA-16-5p (miR-16-5p) plays a protective role in diabetic retinopathy (DR). Its reduction in high glucose conditions promotes retinal cell proliferation and inhibits apoptosis by upregulating VEGFA and TGFBR1.

Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Endocrinology

Background:

  • Diabetic retinopathy (DR) is a leading cause of vision loss in diabetic patients.
  • MicroRNAs (miRNAs) are implicated in the pathogenesis of DR.
  • Understanding the role of specific miRNAs, like miR-16-5p, is crucial for developing targeted therapies.

Purpose of the Study:

  • To investigate the effect of miR-16-5p on high glucose (HG)-induced human retinal microvascular endothelial cells (HRECs).
  • To explore the modulation of vascular endothelial growth factor A (VEGFA) and transforming growth factor beta receptor 1 (TGFBR1) by miR-16-5p in DR.
  • To elucidate the underlying mechanisms of miR-16-5p in regulating HREC proliferation and apoptosis.

Main Methods:

  • Establishment of a DR cell model using HG concentrations (5-30 mM) on HRECs.
  • Quantification of miR-16-5p, VEGFA, and TGFBR1 expression via RT-qPCR and Western blot.
  • Assessment of HREC proliferation using MTT assay and apoptosis using flow cytometry.
  • Validation of miR-16-5p targeting of VEGFA and TGFBR1 using dual-luciferase reporter assays.

Main Results:

  • HG treatment reduced miR-16-5p expression while increasing VEGFA and TGFBR1 mRNA and protein levels in HRECs.
  • Knockdown of miR-16-5p promoted HREC proliferation and inhibited apoptosis under HG conditions.
  • Inhibition of VEGFA and TGFBR1 reversed the effects of miR-16-5p knockdown.
  • Dual-luciferase assays confirmed VEGFA and TGFBR1 as direct targets of miR-16-5p.

Conclusions:

  • miR-16-5p downregulation contributes to DR pathogenesis by promoting HREC proliferation and inhibiting apoptosis.
  • Upregulation of VEGFA and TGFBR1 by reduced miR-16-5p is a key mechanism in HG-induced HREC dysfunction.
  • Restoring miR-16-5p levels or inhibiting VEGFA/TGFBR1 may offer therapeutic strategies for diabetic retinopathy.
Keywords:
TGFBR1VEGFAdiabetic retinopathymiR-16-5pproliferationretinal microvascular endothelial cells

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