Department of Ophthalmology, Affiliated First Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Anhui, 230001, China.
Acta Diabetol. 2022 Mar;59(3):369-380. doi: 10.1007/s00592-021-01813-8. Epub 2021 Oct 30.
Diabetes retinopathy (DR) is associated with retinal microvascular system injury induced by high glucose (HG). This study aims to explore the role and mechanism of long non-coding RNA THRIL in regulating cell proliferation and migration of human retina microvascular endothelial cells (hRMECs) under HG condition.
The gene and protein expression were detetced by RT-PCR and western blot, respectively. Cell proliferation and migration of hRMECs were examined using MTT assay and Transwell assay, respectively. The interaction between miR-125b-5p and THRIL or autophagy-related gene 4D (ATG4D) was analyzed using luciferase activity assay.
THRIL expression was induced by HG in hRMECs. THRIL overexpression enhanced the proliferation and migration of hRMECs induced by HG, whereas THRIL silencing yielded the opposite results. Furthermore, THRIL overexpression induced autophagy activation, and inhibition of autophagy by 3-methyladenine abrogated the promotory effects of THRIL overexpression on cell proliferation and migration of hRMECs. Mechanismly, THRIL inhibited miR-125b-5p to upregulate the expression of ATG4D (an important autophagy-related gene), thereby promoting autophagy. Moreover, miR-125b-5p overexpression or ATG4D silencing alone abolished the promoting effects of THRIL overexpression on HG-induced autophagy, proliferation and migration of hRMECs.
THRIL promotes HG-induced cell proliferation and migration of hRMECs through activation of autophagy via the miR-125b-5p/ATG4D axis. THRIL may serve as a potential therapeutic target for DR.
糖尿病视网膜病变（DR）与高血糖（HG）诱导的视网膜微血管系统损伤有关。本研究旨在探讨长链非编码 RNA THRIL 在调节 HG 条件下人视网膜微血管内皮细胞（hRMEC）增殖和迁移中的作用和机制。
分别通过 RT-PCR 和 Western blot 检测基因和蛋白表达。使用 MTT 测定法和 Transwell 测定法分别检测 hRMEC 细胞的增殖和迁移。通过荧光素酶活性测定分析 miR-125b-5p 与 THRIL 或自噬相关基因 4D（ATG4D）之间的相互作用。
HG 诱导 hRMEC 中 THRIL 的表达。THRIL 过表达增强了 HG 诱导的 hRMEC 增殖和迁移，而 THRIL 沉默则产生了相反的结果。此外，THRIL 过表达诱导自噬激活，而 3-甲基腺嘌呤抑制自噬则消除了 THRIL 过表达对 hRMEC 细胞增殖和迁移的促进作用。机制上，THRIL 抑制 miR-125b-5p 以上调 ATG4D（一种重要的自噬相关基因）的表达，从而促进自噬。此外，miR-125b-5p 过表达或 ATG4D 沉默均可消除 THRIL 过表达对 HG 诱导的 hRMEC 自噬、增殖和迁移的促进作用。
THRIL 通过 miR-125b-5p/ATG4D 轴促进 HG 诱导的 hRMEC 细胞增殖和迁移，从而促进自噬。THRIL 可能成为 DR 的潜在治疗靶点。
