Sun Xincheng, Chen Chen, Liu Hu, Tang Shaowen
Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Department of Ophthalmology, The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu 213003, P.R. China.
Exp Ther Med. 2021 Dec;22(6):1411. doi: 10.3892/etm.2021.10847. Epub 2021 Oct 7.
Diabetic retinopathy, a common complication of diabetes, is the leading cause of blindness globally. Müller cells are key players in diabetes-associated retinal inflammation and dysfunction. However, the pathological changes of Müller cells in response to high glucose (HG) and the underlying mechanism remain unclear. The aim of the present study was to investigate the key role of heat shock protein 47 (HSP47) in HG-induced unfolded protein and inflammatory responses. Primary mouse Müller cells were starved in serum-free DMEM overnight and then treated with HG (30 mM) for 0, 6, 12 or 24 h. It was observed that HG (30 mM) significantly induced the protein expression of HSP47, inositol-requiring transmembrane kinase and endonuclease-1α (IRE1α) and spliced X-box-binding protein 1 (XBP1s) in primary mouse Müller cells compared with the untreated group. In addition, the immunoprecipitation results revealed that HSP47 directly interacted with IRE1α, and this interaction was significantly enhanced by HG exposure for 12 or 24 h compared with the untreated group. Furthermore, small interfering RNA-mediated silencing of HSP47 significantly suppressed HG-induced activation of the IRE1α/XBP1s/hypoxia inducible factor-1 subunit α (HIF-1α) pathway and upregulation of the mRNA expression levels of the inflammatory cytokines vascular endothelial growth factor, platelet-derived growth factor subunit B, inducible nitric oxide synthase and angiopoietin-2 in Müller cells. Furthermore, overexpression of IRE1α or HIF-1α partially attenuated HSP47-siRNA-mediated inhibition of inflammatory cytokine expression in Müller cells. Collectively, these results indicated that HG may induce HSP47 expression and promote the inflammatory response through enhancing the interaction between HSP47 and IRE1α, and activating the IRE1α/XBP1s/HIF-1α pathway in retinal Müller cells.
糖尿病视网膜病变是糖尿病常见的并发症,是全球失明的主要原因。穆勒细胞是糖尿病相关视网膜炎症和功能障碍的关键因素。然而,穆勒细胞对高糖(HG)反应的病理变化及其潜在机制仍不清楚。本研究的目的是探讨热休克蛋白47(HSP47)在HG诱导的未折叠蛋白和炎症反应中的关键作用。原代小鼠穆勒细胞在无血清DMEM中饥饿过夜,然后用HG(30 mM)处理0、6、12或24小时。观察到与未处理组相比,HG(30 mM)显著诱导原代小鼠穆勒细胞中HSP47、肌醇需求跨膜激酶和核酸内切酶-1α(IRE1α)以及剪接的X盒结合蛋白1(XBP1s)的蛋白表达。此外,免疫沉淀结果显示HSP47直接与IRE1α相互作用,与未处理组相比,HG暴露12或24小时后这种相互作用显著增强。此外,小干扰RNA介导的HSP47沉默显著抑制了HG诱导的穆勒细胞中IRE1α/XBP1s/缺氧诱导因子-1亚基α(HIF-1α)途径的激活以及炎症细胞因子血管内皮生长因子、血小板衍生生长因子亚基B、诱导型一氧化氮合酶和血管生成素-2的mRNA表达水平上调。此外,IRE1α或HIF-1α的过表达部分减弱了HSP47-siRNA介导的穆勒细胞中炎症细胞因子表达的抑制。总的来说,这些结果表明HG可能通过增强HSP47与IRE1α之间的相互作用并激活视网膜穆勒细胞中的IRE1α/XBP1s/HIF-1α途径来诱导HSP47表达并促进炎症反应。