Cao Xin, Xue Li-Dan, Di Yue, Li Tao, Tian Ya-Jing, Song Yu
Department of Ophthalmology, The Second Affiliated Hospital of Nantong University, Nantong 226000, Jiangsu Province, PR China.
Department of Ophthalmology, The Second Affiliated Hospital of Nantong University, Nantong 226000, Jiangsu Province, PR China.
Life Sci. 2021 May 1;272:119232. doi: 10.1016/j.lfs.2021.119232. Epub 2021 Feb 16.
Diabetic retinopathy (DR) is the most common complication of type 2 diabetes mellitus, which could result in visual impairment. Accumulating studies have shown the implication of long non-coding RNAs (lncRNAs) in the pathogenesis of DR. Our aims are to investigate whether lncRNA SNHG7 plays a role during DR pathogenesis.
Human retinal microvascular endothelial cells (HRMECs) were treated with high glucose (HG) to build cell model. Relative expression of RNAs were examined using qPCR, and western blot or immunofluorescence analysis was adopted to detect the protein expression. Cell viability, migration and angiogenic capacity of HRMECs were estimated through CCK-8, transwell and tube formation experiments, respectively. Dual-luciferase reporter and RNA pull down assays were employed to verify the interplay between miR-34a-5p and SNHG7 or XBP1. Mesenchymal stem cells (MSCs) were identified by examining typical surface makers using flow cytometry and the differentiation abilities via Alizarin red, Oil red O and Alcian blue staining. MSC-derived exosomes were verified by transmission electron microscopy and western blot.
LncRNA SNHG7 sponged to and negatively regulated miR-34a-5p. SNHG7 overexpression repressed HG induced endothelial-mesenchymal transition (EndMT) and tube formation of HRMECs, while miR-34a-5p overexpression could reverse this effect. miR-34a-5p targeted and negative regulated XBP1. Knockdown of miR-34a-5p repressed HG induced EndMT and tube formation, which were partially blocked by XBP1 inhibition. MSC-derived exosomes could transfer SNHG7 to HRMECs and modulated EndMT and tube formation.
The MSC-derived exosomal lncRNA SNHG7 suppresses EndMT and tube formation in HRMECs via miR-34a-5p/XBP1 axis.
糖尿病视网膜病变（DR）是2型糖尿病最常见的并发症，可导致视力损害。越来越多的研究表明长链非编码RNA（lncRNAs）参与了DR的发病机制。我们的目的是研究lncRNA SNHG7在DR发病过程中是否发挥作用。
用高糖（HG）处理人视网膜微血管内皮细胞（HRMECs）以建立细胞模型。采用qPCR检测RNA的相对表达量，并用蛋白质免疫印迹法或免疫荧光分析法检测蛋白质表达。分别通过CCK-8、Transwell和管腔形成实验评估HRMECs的细胞活力、迁移能力和血管生成能力。采用双荧光素酶报告基因检测和RNA下拉实验验证miR-34a-5p与SNHG7或XBP1之间的相互作用。通过流式细胞术检测典型表面标志物以及茜素红、油红O和阿尔辛蓝染色检测分化能力来鉴定间充质干细胞（MSCs）。通过透射电子显微镜和蛋白质免疫印迹法验证MSC来源的外泌体。
lncRNA SNHG7与miR-34a-5p结合并对其起负向调控作用。SNHG7过表达抑制HG诱导的HRMECs内皮-间充质转化（EndMT）和管腔形成，而miR-34a-5p过表达可逆转这种作用。miR-34a-5p靶向XBP1并对其起负向调控作用。敲低miR-34a-5p可抑制HG诱导的EndMT和管腔形成，而XBP1抑制可部分阻断这种作用。MSC来源的外泌体可将SNHG7转移至HRMECs并调节EndMT和管腔形成。
MSC来源的外泌体lncRNA SNHG7通过miR-34a-5p/XBP1轴抑制HRMECs中的EndMT和管腔形成。
