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Downregulation of glycoprotein non-metastatic melanoma protein B prevents high glucose-induced angiogenesis in diabetic retinopathy

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Abstract

Diabetic retinopathy (DR), a microvascular complication characterized by abnormal angiogenesis, is the most common reason for irreversible blindness. Glycoprotein non-metastatic melanoma protein B (GPNMB), as a transmembrane protein, was found to be associated with angiogenesis. This study aims to investigate the role of GPNMB in DR. The levels of GPNMB and Integrin β1 were detected by real-time PCR and western blot and were found to be increased in human retinal microvascular endothelial cells (HRMECs) with high glucose (HG, 25 mmol/L) treatment. Knockdown of GPNMB was mediated by lentivirus carrying shRNA targeting GPNMB in vivo and in vitro. Functional experiments, including cell counting kit-8 (CCK-8), scratch, and tube formation assays, showed the anti-proliferative, anti-migrative, and anti-angiogenic roles of GPNMB knockdown in HRMECs using the lentivirus system following HG challenge. Additionally, increased GPNMB levels were detected in the retina of DR rats induced by a single intraperitoneal injection of streptozotocin (60 mg/kg) using real-time PCR, western blot, and immunofluorescence assays. Downregulation of GPNMB inhibited the angiogenesis and vascular endothelial growth factor production in the retina of rats with DR. Furthermore, overexpression of Integrin β1 led to increased angiogenesis in DR. Integrin β1 was indicated as a target protein of GPNMB. Upregulated-Integrin β1 restored GPNMB knockdown-induced inhibition of cell viability, migration, and tube formation in HRMECs. In conclusion, we provide evidence for the angiogenic role of GPNMB and demonstrate that silencing GPNMB may represent a therapeutic potential in the treatment of DR.

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Data availability

Data in this study is available from the corresponding author.

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Funding

This study was supported by the Key Scientific Research Project of Colleges and Universities in Henan Province (19A320059).

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Authors and Affiliations

  1. Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, No. 1, Jianshe East Road, Zhengzhou, Henan, People’s Republic of China

    Tingyu Qin, Xiangying Xi & Zhipeng Wu

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  2. Xiangying Xi
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Contributions

QT: designed and carried out the experiments, performed data analysis and wrote the manuscript. XX: carried out the experiments and revised the manuscript. WZ: performed data analysis and revised the manuscript.

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Correspondence to Tingyu Qin.

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All procedures in this study were performed in agreement with the Ethics Committee in The First Affiliated Hospital of Zhengzhou University.

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Qin, T., Xi, X. & Wu, Z. Downregulation of glycoprotein non-metastatic melanoma protein B prevents high glucose-induced angiogenesis in diabetic retinopathy. Mol Cell Biochem 478, 697–706 (2023). https://doi.org/10.1007/s11010-022-04537-7

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  • DOI: https://doi.org/10.1007/s11010-022-04537-7

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