目的
糖尿病性视网膜病（DR）是糖尿病的血管并发症，可导致视力障碍和失明。长非编码RNA（lncRNA）已被发现参与包括DR在内的多种疾病的生物学过程。我们设计了本研究，以研究TPTEP1在DR中的特定作用。
方法
首先，我们通过高葡萄糖（HG）刺激人视网膜血管内皮细胞（HRVEC）来模拟糖尿病，并使用RT-qPCR分析测量HG刺激的HRVECs中TPTEP1的表达。然后，进行了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，通过生物信息学分析，RNA下拉，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HRVECs中TPTEP1，转录信号转导和转录激活因子3（STAT3）与血管内皮生长因子A（VEGFA）的关系。 。最后，为了验证HG刺激的HRVEC中TPTEP1 / STAT3 / VEGFA轴的调控，在HG刺激的HRVEC中进行了抢救实验。
结果
TPTEP1在HG刺激的HRVEC中表现出明显的下调。此外，TPTEP1过表达降低了HG刺激的HRVECs的活力，迁移和血管生成。此外，TPTEP1抑制STAT3的磷酸化和核易位，从而下调VEGFA mRNA和蛋白水平。此外，STAT3上调或VEGFA上调抵消了TPTEP1过表达介导的HRVECs中HG诱导的功能障碍的抑制。
结论
TPTEP1通过与STAT3相互作用并靶向VEGFA减轻了HRVEC中HG引起的功能障碍。
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.
