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<p><i>Diabetic</i> <i>retinopathy</i> (<i>DR</i>) is a microvascular complication of diabetes. <i>Insulin-like growth factor 1 receptor</i> (<i>IGF1R</i>) has been implicated in the pathogenesis of <i>DR</i>; however, the underlying mechanism remains unclear. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess <i>IGF1R</i> mRNA expression. Western blotting assays were performed to analyze the protein expression of <i>IGF1R</i>, <i>ubiquitin-specific peptidase 14</i> (<i>USP14</i>), and <i>BRCA1-associated protein 1</i> (<i>BAP1</i>). Cell viability, apoptosis, interleukin-1 beta (IL-1β), and <i>tumor necrosis factor-alpha</i> (<i>TNF-α</i>) levels were analyzed using cell counting kit-8 assay, flow cytometry, and enzyme-linked immunosorbent assays, respectively. Fluorescent microscopy and flow cytometry were performed for <i>reactive oxygen species</i> (<i>ROS</i>) level assessment, and colorimetric assays for <i>iron</i> (<i>Fe</i><sup><i>2</i>+</sup>) and <i>glutathione</i> (<i>GSH</i>) levels. Co-immunoprecipitation assays and/or colocalization techniques were employed to validate the association of <i>IGF1R</i> with <i>USP14</i> and <i>BAP1</i>. Treatment with high glucose (HG) increased the protein expression of <i>IGF1R</i>, <i>USP14</i>, and <i>BAP1</i> in <i>ARPE-19</i> cells. Silencing of <i>IGF1R</i> mitigated HG-induced apoptosis, inflammatory response, and ferroptosis in <i>ARPE-19</i> cells. <i>USP14</i> was found to stabilize <i>IGF1R</i> protein expression through deubiquitination. Overexpression of <i>USP14</i> exacerbated HG-induced cellular injury, whereas silencing of <i>USP14</i> protected <i>ARPE-19</i> cells by reducing <i>IGF1R</i> expression. Interaction between <i>IGF1R</i> and <i>BAP1</i> was confirmed in <i>ARPE-19</i> cells and <i>IGF1R</i> silencing protected cells from HG-induced injury by regulating <i>BAP1</i> expression. Thus, <i>USP14</i>-dependent regulation of <i>IGF1R</i> expression and its interaction with <i>BAP1</i> play a crucial role in the pathogenesis of high glucose-induced <i>diabetic retinopathy</i>.</p> Graphical Abstract <p></p>

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USP14-Dependent IGF1R Aggravates High Glucose-Induced Diabetic Retinopathy by Upregulating BAP1

  • 期刊论文

  • 发表日期：

    2025年3月31日

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  • Li Yu,
  • Xia Zheng,
  • Yan Wu,
  • Kui Ge

摘要

Diabetic retinopathy (DR) is a microvascular complication of diabetes. Insulin-like growth factor 1 receptor (IGF1R) has been implicated in the pathogenesis of DR; however, the underlying mechanism remains unclear. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to assess IGF1R mRNA expression. Western blotting assays were performed to analyze the protein expression of IGF1R, ubiquitin-specific peptidase 14 (USP14), and BRCA1-associated protein 1 (BAP1). Cell viability, apoptosis, interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) levels were analyzed using cell counting kit-8 assay, flow cytometry, and enzyme-linked immunosorbent assays, respectively. Fluorescent microscopy and flow cytometry were performed for reactive oxygen species (ROS) level assessment, and colorimetric assays for iron (Fe2+) and glutathione (GSH) levels. Co-immunoprecipitation assays and/or colocalization techniques were employed to validate the association of IGF1R with USP14 and BAP1. Treatment with high glucose (HG) increased the protein expression of IGF1R, USP14, and BAP1 in ARPE-19 cells. Silencing of IGF1R mitigated HG-induced apoptosis, inflammatory response, and ferroptosis in ARPE-19 cells. USP14 was found to stabilize IGF1R protein expression through deubiquitination. Overexpression of USP14 exacerbated HG-induced cellular injury, whereas silencing of USP14 protected ARPE-19 cells by reducing IGF1R expression. Interaction between IGF1R and BAP1 was confirmed in ARPE-19 cells and IGF1R silencing protected cells from HG-induced injury by regulating BAP1 expression. Thus, USP14-dependent regulation of IGF1R expression and its interaction with BAP1 play a crucial role in the pathogenesis of high glucose-induced diabetic retinopathy.

Graphical Abstract