TGF-β1 Mediates the EndoMt in High Glucose-Treated Human Retinal Microvascular Endothelial Cells

ABSTRACT

The purpose of our study was to investigate the role of TGF-β1 in the endothelial-to-mesenchymal transition (EndoMT) and fibrosis in high glucose (HG)-treated human retinal microvascular endothelial cells (HRMECs). HRMECs were cultured not only under normal glucose (NG) conditions with or without TGF-β1, but also under HG conditions with or without the TGF-β1 inhibitor SB431542. The expression of TGF-β1 was detected by real time-PCR and enzyme-linked immunosorbent assay. Morphological changes and migration of the HRMECs were observed using electron microscopy and scratch-wound assay. Endothelial markers, such as CD31 and vascular endothelial (VE)-cadherin, and the acquisition of fibrotic markers, such as alpha smooth muscle actin (α-SMA) and fibroblast-specific protein-1 (FSP-1), were determined by immunofluorescent staining and western blot. The level of TGF-β1 was significantly upregulated in HG-treated HRMECs. And HG stimulation promoted obvious morphological changes and the migration ability in HRMECs. Our results also demonstrated increased expression of α-SMA and FSP-1, and decreased expression of CD31 and VE-cadherin, in HG-treated HRMECs. These EndoMT-related changes were promoted by TGF-β1 and abrogated by SB431542. The results of this study demonstrated the important role of TGF-β1 in HG-induced vitreoretinal fibrosis. EndoMT is likely to be involved in the associated effects.

KEYWORDS:

• Diabetic retinopathy
• EndoMT
• Hrmecs
• TGF-β1
• Vitreoretinal fibrosis

Acknowledgments

This manuscript has been edited and proofread by Textcheck Bioscience Limited.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

JL conceived and designed the experiments. JY and PP W performed the experiments and wrote the manuscript. XH and XJ performed the statistical analysis. All authors gave final approval of the version to be published.

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Ethics approval and consent to participate

All experiments conformed to internationally accepted standards and were approved by the appropriate institutional review body.

Additional information

Funding

The current study was supported by Health Development Planning Commission Science Foundation of Zhejiang Province (Grant Nos 2020364455, 2022ky1434).