Targeting long noncoding RNA-AQP4-AS1 for the treatment of retinal neurovascular dysfunction in diabetes mellitus.
Targeting long noncoding RNA-AQP4-AS1 for the treatment of retinal neurovascular dysfunction in diabetes mellitus.
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靶向长非编码RNA-AQP4-AS1治疗糖尿病视网膜神经血管功能障碍
DOI：
10.1016/j.ebiom.2022.103857
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发表时间：
2022-03
期刊：
影响因子：
11.1
通讯作者：
Jiang Q
中科院分区：
文献类型：
--
作者：
Li X;Zhu J;Zhong Y;Liu C;Yao M;Sun Y;Yao W;Ni X;Zhou F;Yao J;Jiang Q
糖尿病视网膜病变(DR)是导致工作年龄人群失明的主要原因，其特征是视网膜神经变性和血管功能障碍。长非编码RNA(LncRNAs)已成为多种生物学过程和疾病进展的重要调节因子。在这里，我们研究了lncRNA AQP4-AS1在糖尿病引起的视网膜神经血管功能障碍中的作用。定量RT-PCR检测糖尿病应激时AQP4-AS1的表达模式。采用视觉电生理、TUNEL染色、伊文思蓝染色、视网膜胰酶消化和免疫荧光染色等方法检测AQP4-AS1在体内视网膜神经血管功能障碍中的作用。采用四甲基偶氮唑盐比色法、原位缺口末端标记法、PI/Calcein-AM染色、EDU掺入实验、Transwell实验和小管形成实验检测AQP4-AS1对体外培养的视网膜细胞功能的影响。QRT-PCR、Western印迹和体内研究揭示了AQP4-AS1介导的视网膜神经血管功能障碍的机制。AQP4-AS1在糖尿病视网膜病变患者、高糖处理的Müller细胞和糖尿病小鼠模型视网膜的临床样本中显著增加。体内沉默AQP4-AS1可减轻视网膜神经退行性变和血管功能障碍，表现为改善视网膜毛细血管变性，减少反应性胶质增生，减少RGC丢失。AQP4-AS1在体外直接调节Müler细胞功能，间接影响内皮细胞和RGC功能。机制上，AQP4-AS1通过影响AQP4水平调节视网膜神经血管功能障碍。本研究得到了中国国家自然科学基金(批准号：81800858、82070983、81870679、81970823)、南京市医学科技发展专项基金(批准号：ZKX17053、YKK19158)、江苏省创新团队项目基金(批准号：：和南京市科技发展计划项目基金(批准号：201716007、201805007和201803058)。
Diabetic retinopathy (DR) is a leading cause of blindness in the working-age population, which is characterized by retinal neurodegeneration and vascular dysfunction. Long non-coding RNAs (LncRNAs) have emerged as critical regulators in several biological processes and disease progression. Here we investigated the role of lncRNA AQP4-AS1 in retinal neurovascular dysfunction induced by diabetes. Quantitative RT-PCR was used to detect the AQP4-AS1 expression pattern upon diabetes mellitus-related stresses. Visual electrophysiology examination, TUNEL staining, Evans blue staining, retinal trypsin digestion and immunofluorescent staining were conducted to detect the role of AQP4-AS1 in retinal neurovascular dysfunction in vivo. MTT assays, TUNEL staining, PI/Calcein-AM staining, EdU incorporation assay transwell assay and tube formation were conducted to detect the role of AQP4-AS1 in retinal cells function in vitro. qRT-PCR, western blot and in vivo studies were conducted to reveal the mechanism of AQP4-AS1-mediated retinal neurovascular dysfunction. AQP4-AS1 was significantly increased in the clinical samples of diabetic retinopathy patients, high glucose-treated Müller cells, and diabetic retinas of a murine model. AQP4-AS1 silencing in vivo alleviated retinal neurodegeneration and vascular dysfunction as shown by improved retinal capillary degeneration, decreased reactive gliosis, and reduced RGC loss. AQP4-AS1 directly regulated Müller cell function and indirectly affected endothelial cell and RGC function in vitro. Mechanistically, AQP4-AS1 regulated retinal neurovascular dysfunction through affecting AQP4 levels. This study reveals AQP4-AS1 is involved in retinal neurovascular dysfunction and expected to become a promising target for the treatment of neurovascular dysfunction in DR. This work was generously supported by the grants from the National Natural Science Foundation of China (Grant No. 81800858, 82070983, 81870679 and 81970823), grants from the Medical Science and Technology Development Project Fund of Nanjing (Grant No ZKX17053 and YKK19158), grants from Innovation Team Project Fund of Jiangsu Province (No. CXTDB2017010), and the Science and Technology Development Plan Project Fund of Nanjing (Grant No 201716007, 201805007 and 201803058).
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