Yang Ying
Department of Ophthalmology, Sichuan Provincial People's Hospital, Chengdu, China; School of Medicine, University of Electronic Science and Technology of China, Chengdu, China.
Tissue Cell. 2025 Feb;92:102620. doi: 10.1016/j.tice.2024.102620. Epub 2024 Nov 17.
Diabetic retinopathy (DR) has been proven to be a leading cause of blindness. This study aimed to investigate the effect of Yes-associated protein 1 (YAP1) on the hypoxia-induced DR mice retinal microvascular endothelial cells (MRMECs) model. The hypoxia-induced DR MRMECs model was generated by treating in hypoxia circumstance (5 % CO and 3 % O) for 48 h. This study constructed YAP1 overexpression and taurine-upregulated gene 1 (TUG1) silencing lentiviral vectors, both of which were used to infect the DR MRMECs model. Quantitative real-time PCR (qRT-PCR) was used to amplify the YAP1, TUG1, vascular endothelial growth factor A (VEGFA), and miR-144-3p gene. Western blot was used to identify the expression of YAP1 and VEGFA. The CCK-8 assay was used to evaluate proliferation and the flow cytometry assay was used to determine apoptosis of MRMECs. Cell migration and tube formation were also evaluated. The results showed that YAP1 overexpression and TUG1 silencing lentivirus were successfully constructed. YAP1 overexpression significantly promoted, but TUG1 silence inhibited cell proliferation and migration compared to DR MRMECs model (P<0.05). YAP1 markedly promoted TUG1/VEGFA and reduced miR-144-3p gene transcription compared to those of the DR MRMECs model (P<0.05). YAP1 overexpression and TUG1 silence demonstrated the opposite effects on VEGFA expression. YAP1 overexpression obviously promoted tube formation of MRMECs. In conclusion, overexpression of YAP1 promoted cell proliferation, cell migration, TUG1 and VEGFA expression, and reduced the transcription of the miR-144-3p gene in DR MRMECs. Overexpression of YAP1 aggravated the progress of DR in MRMECs by activating the TUG1/miR-144-3p/VEGFA signaling pathway.
糖尿病视网膜病变（DR）已被证明是导致失明的主要原因。本研究旨在探讨Yes相关蛋白1（YAP1）对缺氧诱导的DR小鼠视网膜微血管内皮细胞（MRMECs）模型的影响。通过在缺氧环境（5%二氧化碳和3%氧气）中处理48小时构建缺氧诱导的DR MRMECs模型。本研究构建了YAP1过表达和牛磺酸上调基因1（TUG1）沉默慢病毒载体，二者均用于感染DR MRMECs模型。采用定量实时PCR（qRT-PCR）扩增YAP1、TUG1、血管内皮生长因子A（VEGFA）和miR-144-3p基因。采用蛋白质免疫印迹法检测YAP1和VEGFA的表达。采用CCK-8法评估增殖情况，采用流式细胞术检测MRMECs的凋亡情况。同时评估细胞迁移和管腔形成情况。结果显示成功构建了YAP1过表达和TUG1沉默慢病毒。与DR MRMECs模型相比，YAP1过表达显著促进细胞增殖和迁移，但TUG1沉默则抑制细胞增殖和迁移（P<0.05）。与DR MRMECs模型相比，YAP1显著促进TUG1/VEGFA表达并降低miR-144-3p基因转录（P<0.05）。YAP1过表达和TUG1沉默对VEGFA表达具有相反的影响。YAP1过表达明显促进MRMECs的管腔形成。总之，YAP1过表达促进了DR MRMECs的细胞增殖、细胞迁移、TUG1和VEGFA表达，并降低了miR-144-3p基因的转录。YAP1过表达通过激活TUG1/miR-144-3p/VEGFA信号通路加重了MRMECs中DR的进展。
