Aims

Diabetic retinopathy (DR) is a vascular complication of diabetes mellitus that causes visual impairment and blindness. Long noncoding RNAs (lncRNAs) have been revealed to be involved in biological processes of several diseases including DR. We designed this study to investigate the specific role of TPTEP1 in DR.

Methods

First, we mimicked diabetic conditions with high glucose (HG) stimulation of human retinal vascular endothelial cells (HRVECs) and measured TPTEP1 expression in HG-stimulated HRVECs using RT-qPCR analysis. Then, CCK-8, Transwell, and Matrigel tube formation assays as well as western blot analysis were performed to reveal the biological functions of TPTEP1 in HG-stimulated HRVECs. Subsequently, bioinformatics analysis, RNA pull down, luciferase reporter and ChIP assays as well as western blot analysis evaluated the relationship of TPTEP1, signal transducer and activator of transcription 3 (STAT3) and vascular endothelial growth factor A (VEGFA) in HG-stimulated HRVECs. Finally, to verify the regulation of the TPTEP1/STAT3/VEGFA axis in HG-stimulated HRVECs, rescue experiments were carried out in HG-stimulated HRVECs.

Results

TPTEP1 presented a significant downregulation in HG-stimulated HRVECs. Additionally, TPTEP1 overexpression reduced viability, migration, and angiogenesis in HG-stimulated HRVECs. Moreover, TPTEP1 suppressed phosphorylation and nuclear translocation of STAT3, and thereby downregulated VEGFA mRNA and protein levels. Furthermore, TPTEP1 overexpression-mediated suppression of HG-induced dysfunction in HRVECs was countervailed by STAT3 upregulation or VEGFA upregulation.

Conclusions

TPTEP1 alleviated HG-induced dysfunction in HRVECs via interacting with STAT3 and targeting VEGFA.

中文翻译：

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TPTEP1 通过与 STAT3 相互作用并靶向 VEGFA 抑制高糖诱导的视网膜血管内皮细胞功能障碍

宗旨

糖尿病视网膜病变 (DR) 是糖尿病的血管并发症，可导致视力障碍和失明。已发现长链非编码 RNA (lncRNA) 参与包括 DR 在内的多种疾病的生物学过程。我们设计这项研究是为了调查 TPTEP1 在 DR 中的具体作用。

方法

首先，我们用高葡萄糖 (HG) 刺激人视网膜血管内皮细胞 (HRVEC) 模拟糖尿病状况，并使用 RT-qPCR 分析测量 HG 刺激的 HRVEC 中的 TPTEP1 表达。然后，进行 CCK-8、Transwell 和 Matrigel 管形成测定以及蛋白质印迹分析，以揭示 TPTEP1 在 HG 刺激的 HRVEC 中的生物学功能。随后，生物信息学分析、RNA pull down、荧光素酶报告基因和 ChIP 分析以及蛋白质印迹分析评估了 HG 刺激的 HRVEC 中 TPTEP1、信号转导和转录激活因子 3 (STAT3) 和血管内皮生长因子 A (VEGFA) 的关系. 最后，为了验证 HG 刺激的 HRVEC 中 TPTEP1/STAT3/VEGFA 轴的调节，在 HG 刺激的 HRVEC 中进行了拯救实验。

结果

TPTEP1 在 HG 刺激的 HRVEC 中表现出显着下调。此外，TPTEP1 过表达降低了 HG 刺激的 HRVEC 的活力、迁移和血管生成。此外，TPTEP1 抑制 STAT3 的磷酸化和核转位，从而下调 VEGFA mRNA 和蛋白水平。此外，TPTEP1 过表达介导的对 HG 诱导的 HRVEC 功能障碍的抑制被 STAT3 上调或 VEGFA 上调所抵消。

结论

TPTEP1 通过与 STAT3 相互作用并靶向 VEGFA 来减轻 HG 诱导的 HRVEC 功能障碍。