Department of Ophthalmology, Lishui Municipal Central Hospital, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, PR China.
Department of Stomatology, Lishui Municipal Central Hospital, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, PR China.
Semin Ophthalmol. 2024 May;39(4):312-319. doi: 10.1080/08820538.2023.2300806. Epub 2024 Jan 8.
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.
我们的研究目的是探讨 TGF-β1 在高糖（HG）处理的人视网膜微血管内皮细胞（HRMECs）中的内皮到间充质转化（EndoMT）和纤维化中的作用。我们不仅在正常葡萄糖（NG）条件下培养 HRMECs，还在 HG 条件下培养 HRMECs，同时分别加入或不加入 TGF-β1 抑制剂 SB431542。通过实时 PCR 和酶联免疫吸附试验检测 TGF-β1 的表达。通过电子显微镜和划痕实验观察 HRMECs 的形态变化和迁移。通过免疫荧光染色和 Western blot 测定内皮标志物，如 CD31 和血管内皮（VE）-钙黏蛋白，以及纤维化标志物，如α平滑肌肌动蛋白（α-SMA）和成纤维细胞特异性蛋白-1（FSP-1）的获得。结果表明，HG 处理的 HRMECs 中 TGF-β1 的表达明显上调。HG 刺激促进了 HRMECs 中明显的形态变化和迁移能力。我们的结果还表明，HG 处理的 HRMECs 中 α-SMA 和 FSP-1 的表达增加，CD31 和 VE-cadherin 的表达减少。这些与 EndoMT 相关的变化是由 TGF-β1 促进的，并且被 SB431542 阻断。这项研究的结果表明 TGF-β1 在 HG 诱导的玻璃体内视网膜纤维化中起着重要作用。EndoMT 可能参与了相关的影响。
