长链非编码RNA AQP4-AS1靶向Müller细胞反应胶质化调控糖尿病视网膜微血管病变

Diabetic retinopathy is a leading cause of blindness, which is characterized by microvascular pathology, Müller cell gliotic response and retinal ganglion cell apoptosis. It has been shown that Müller cell gliotic response occurs earlier than microvascular changes, yet the underlying mechanism is still not fully understood. lncRNAs have emerged as novel regulatory components of eukaryotic genomes and key players in numerous physiologic and pathological processes. Our preliminary results have revealed that lncRNA AQP4-AS1 is mainly expressed in the Müller cells，and AQP4-AS1 knockdown could decrease gliotic response and microvascular dysfunction. We speculate that AQP4-AS1 could affect the gliotic response, which subsequently affect microvascular dysfunction during the progress of diabetic retinopathy. In this project, we will apply the diabetic model in vitro or in vivo to detect AQP4-AS1 expression pattern in Müller cells, clarify the role of AQP4-AS1 in Müller cell gliosis and microvascular dysfunction through in vitro or in vivo intervention of AQP4-AS1 expression, and reveal the molecular mechanism of AQP4-AS1-mediated Müller dysfunction. The potential results will provide novel targets for the prevention and treatment of diabetic-induced microvascular dysfunction.

糖尿病视网膜病变（Diabetic retinopathy，DR）是重要的致盲性眼病，其病理表现包括微血管病变、Müller细胞反应胶质化和神经节细胞凋亡等。最近研究发现，Müller细胞反应胶质化是最早发生的病理改变，其病变机制尚不完全清楚。长链非编码RNA（lncRNAs）是一类重要的基因元件，参与调控众多生理和病理过程。我们预实验发现，lncRNA-AQP4-AS1主要表达于Müller胶质细胞中，其表达干预后影响糖尿病性反应胶质化和微血管病变的进程。我们推测，AQP4-AS1有望靶向视网膜反应胶质化，进而调控视网膜血管病变进程。本项目拟构建离体和在体糖尿病模型，分析AQP4-AS1的表达规律；基于AQP4-AS1表达干预，明确AQP4-AS1对反应胶质化和微血管病变的调控作用；基于生物信息学和分子生物学技术，阐明AQP4-AS1调控DR的分子机制。研究有望为防治DR提供新的治疗靶点。

糖尿病性视网膜病变(DR)作为糖尿病常见的并发症之一，病变过程中可导致神经系统和血管系统功能紊乱，其已成为全世界致盲的主要原因。本研究旨在探究长链非编码RNA(Long non-coding RNA) AQP4-AS1在糖尿病性视网膜病变神经血管功能障碍中的调控作用以及分子机制。qRT-PCR检测发现，AQP4-AS1在糖尿病小鼠视网膜中表达异常增加，在Müller细胞高糖以及H2O2刺激下表达异常增加。在体实验发现，AQP4-AS1表达沉默可以抑制病理性血管形成，缓解视网膜胶质化以及神经节细胞的凋亡。离体实验发现，AQP4-AS1表达沉默降低了Müller细胞的活性，线粒体膜电位以及增殖能力，促进了细胞凋亡，共培养实验表明，AQP4-AS1表达沉默间接调节视网膜内皮和神经细胞的功能。机制研究发现，AQP4-AS1表达沉默下调了视网膜中VEGF, Ang-1, IL-6, IL-1β和ICAM-1细胞因子的表达同时AQP4-AS1选择性结合AQP4 mRNA，增加了AQP4的表达，从而调控糖尿病诱导的神经血管病变过程。长链非编码RNA AQP4-AS1能够参与调节视网膜神经血管病变过程，AQP4-AS1沉默导致AQP4表达增加，从而减轻糖尿病所致的神经血管功能障碍，AQP4-AS1有望成为治疗糖尿病诱导的视网膜神经血管病变的新靶点。