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

IF 3.1 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Li Yu, Xia Zheng, Yan Wu, Kui Ge
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引用次数: 0

Abstract

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.

usp14依赖性IGF1R通过上调BAP1加重高糖诱导的糖尿病视网膜病变
糖尿病视网膜病变(DR)是糖尿病的微血管并发症。胰岛素样生长因子1受体(IGF1R)与糖尿病的发病机制有关;然而,其潜在机制尚不清楚。采用定量反转录聚合酶链反应(qRT-PCR)检测IGF1R mRNA的表达。Western blotting检测IGF1R、泛素特异性肽酶14 (USP14)和brca1相关蛋白1 (BAP1)的蛋白表达。分别采用细胞计数试剂盒-8、流式细胞术和酶联免疫吸附法分析细胞活力、凋亡、白细胞介素-1β (IL-1β)和肿瘤坏死因子-α (TNF-α)水平。荧光显微镜和流式细胞术检测活性氧(ROS)水平,比色法检测铁(Fe2+)和谷胱甘肽(GSH)水平。采用共免疫沉淀法和/或共定位技术验证IGF1R与USP14和BAP1的关联。高糖(HG)处理增加了ARPE-19细胞中IGF1R、USP14和BAP1的蛋白表达。IGF1R沉默可减轻hg诱导的ARPE-19细胞凋亡、炎症反应和铁下垂。发现USP14通过去泛素化稳定IGF1R蛋白的表达。USP14的过表达加重了hg诱导的细胞损伤,而USP14的沉默通过降低IGF1R的表达来保护ARPE-19细胞。在ARPE-19细胞中证实了IGF1R和BAP1的相互作用,IGF1R通过调节BAP1的表达来保护细胞免受hg诱导的损伤。因此,依赖usp14的IGF1R表达调控及其与BAP1的相互作用在高糖诱导的糖尿病视网膜病变的发病机制中起着至关重要的作用。
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来源期刊
Applied Biochemistry and Biotechnology
Applied Biochemistry and Biotechnology 工程技术-生化与分子生物学
CiteScore
5.70
自引率
6.70%
发文量
460
审稿时长
5.3 months
期刊介绍: This journal is devoted to publishing the highest quality innovative papers in the fields of biochemistry and biotechnology. The typical focus of the journal is to report applications of novel scientific and technological breakthroughs, as well as technological subjects that are still in the proof-of-concept stage. Applied Biochemistry and Biotechnology provides a forum for case studies and practical concepts of biotechnology, utilization, including controls, statistical data analysis, problem descriptions unique to a particular application, and bioprocess economic analyses. The journal publishes reviews deemed of interest to readers, as well as book reviews, meeting and symposia notices, and news items relating to biotechnology in both the industrial and academic communities. In addition, Applied Biochemistry and Biotechnology often publishes lists of patents and publications of special interest to readers.
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