has_circ_0000371靶向调控miR-181c/FoxO1轴在糖尿病视网膜病变微血管内皮细胞损伤中的机制研究

Diabetic retinopathy is characteristic of progressive dysfunction of retinal microvascular, which was recently found to be related to abnormal circular RNA regulation, but the specific mechanism is still unclear. In the pilot study of this project, intraocular tissues of PDR and NDM patients were used for RNA sequence analysis. It was found that circ_0000371 was significantly up-regulated in the PDR group with down regulation of miR-181c and greatly increased expression of FoxO1 mRNA. Both miR-181c and FoxO1 mRNA have already been confirmed involving the pathogenesis of DR microvascular dysfunction. Based on the previous researches and our pilot study, we hypothesized that circ_0000371 would regulate the microvascular function of retina under high glucose by miR-181c/FoxO1 axis and downstream molecular. Vitro and vivo experiments will be utilized to investigate the influence of circ_0000371 on cell function, cell and retina molecular expression, retina function and vascular morphology under high glucose. Luciferase reporter gene system will be used to validate the regulation network among circ_0000371, miR-181c and FoxO1. Discovering the molecular mechanism of circ_0000371 on retinal vascular endothelial function will provide a theoretical basis for the new strategy of diabetic retinopathy.

环状RNA（circRNA）在糖尿病视网膜病变（DR）微血管内皮细胞损伤中的生物学作用亟待阐明。申请人前期高通量RNA-seq筛选和低通量qPCR验证，鉴定出circ_0000371在DR眼组织样本中显著上调；生物信息学分析显示，circ_0000371与miR-181c存在紧密结合位点；并证实DR样本中FoxO1上调、miR-181c下调，circ_0000371、FoxO1、miR-181c三者表达趋势符合“海绵机制”模式。已知miR-181c通过FoxO1介导视网膜微血管内皮细胞损伤。因此，可能存在“circ_0000371/miR-181c/FoxO1/内皮损伤”环路。拟从细胞、动物水平，探讨circ_0000371作为miR-181c海绵调控FoxO1表达介导视网膜微血管内皮细胞损伤的机制。本研究将从circRNA转录后调控这一新视角丰富对DR机制的阐释，并为DR防治提供新思路。

糖尿病视网膜病变(DR)缺乏早期检测的生物标志物，新发现的circRNA似乎是新型分子标记和治疗靶标的理想候选者。本项目聚焦于circRNA在DR中的潜在作用和机制，采用标准荟萃分析方法，系统总结了circRNA调控糖尿病视网膜病变发生的功能和机制，揭示在DR发生发展过程中，circRNA转录后调控是重要的环节；建立糖尿病患者数据库，高度近视也与VEGF、IL等因子密切相关，通过分析多模态数据，发现DR与高度近视重要表型独立相关，可能与眼轴增长引发的视网膜的低血流灌注状态、VEGF等细胞因子的调节作用有关。通过高通量分析和低通量验证，发现VEGFA、VEGFB、IL6、CXCL-1、IGF-1明显上升，与circ_0000371相关。衰老和肾功能也与微循环及炎症密切相关，是开发circRNA生物标志物或者治疗靶点时需要考虑的混杂因素。本项目为探索糖尿病视网膜病变微循环损伤机制提供了新思路。项目研究成果在Acta Diabetol等业内高水平期刊发表论文6篇。