MALAT1通过靶向miR-126-5p调控增殖性糖尿病视网膜病变的机制
Mechanisms of MALAT1 regulating proliferative diabetic retinopathy via targeting miR-126-5p.
作者信息
Zhao Wei, Liu Yanli, Li Cairui, Yin Zhijian, Liu Yi, An Meixia
机构信息
The Third Affiliated Hospital, Southern Medical University Guangzhou 510630, Guangdong, China.
Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases Guangzhou 510630, Guangdong, China.
出版信息
Am J Transl Res. 2023 May 15;15(5):3279-3289. eCollection 2023.
OBJECTIVE
Diabetic retinopathy (DR) is the primary reason for blindness among the middle-aged and elderly. It can progress to proliferative diabetic retinopathy (PDR), a condition that is accompanied by retinal neovascularization as the disease worsens. Understanding the pathogenesis of PDR can facilitate the development of treatments. In this study, we aimed to investigate the involvement in the lncRNA MALAT1 (MALAT1)/miR-126-5p axis in modulating PDR progression.
METHODS
Rat retinal endothelial cells (RECs) was induced with 30 mM glucose to build an PDR model. MALAT1 was down-regulated using siRNA sequences, and miR-126-5p was up-regulated with miRNA mimics. Dual-luciferase reporter assay and RNA immunoprecipitation assay were carried out to identify and validate the targeting relationship between MALAT1 and miR-126-5p. Angiogenesis, cell proliferation and cell migration were detected using tubule formation, CCK-8, and scratch assays, respectively. Western blots quantified angiogenesis- and migration-associated genes vascular endothelial growth factor (VEGF), MMP2 and MMP9, while qPCR measured MALAT1 and miR-126-5p levels.
RESULTS
In high-glucose induced RECs, MALAT1 was up-regulated while miR-126-5p was down-regulated. The angiogenesis as well as the proliferation and migration capacities of high glucose-induced RECs were suppressed when MALAT1 was down-regulated or miR-126-5p was up-regulated, accompanied by reductions in VEGF, MMP-2 and MMP9. RNA immunoprecipitation assay confirmed that miR-126-5p could be enriched in MALAT1 sequences. Dual-luciferase reporter assay confirmed the targeted inhibition of miR-126-5p by MALAT1. Downregulating miR-126-5p counteracted the effect of MALAT1 downregulation on RECs induced by high glucose.
CONCLUSIONS
MALAT1 promotes PDR by inhibiting miR126-5p and inducing REC proliferation, migration and angiogenesis.
目的
糖尿病视网膜病变（DR）是中老年人群失明的主要原因。它可进展为增殖性糖尿病视网膜病变（PDR），随着病情加重会出现视网膜新生血管形成。了解PDR的发病机制有助于开发治疗方法。在本研究中，我们旨在探讨长链非编码RNA MALAT1（MALAT1）/miR-126-5p轴在调节PDR进展中的作用。
方法
用30 mM葡萄糖诱导大鼠视网膜内皮细胞（RECs）构建PDR模型。使用小干扰RNA序列下调MALAT1，并使用miRNA模拟物上调miR-126-5p。进行双荧光素酶报告基因检测和RNA免疫沉淀检测，以鉴定和验证MALAT1与miR-126-5p之间的靶向关系。分别使用小管形成、CCK-8和划痕试验检测血管生成、细胞增殖和细胞迁移。蛋白质印迹法定量检测血管生成和迁移相关基因血管内皮生长因子（VEGF）、基质金属蛋白酶2（MMP2）和基质金属蛋白酶9（MMP9），而定量聚合酶链反应（qPCR）检测MALAT1和miR-126-5p水平。
结果
在高糖诱导的RECs中，MALAT1上调而miR-126-5p下调。当MALAT1下调或miR-126-5p上调时，高糖诱导的RECs的血管生成以及增殖和迁移能力受到抑制，同时VEGF、MMP-2和MMP9减少。RNA免疫沉淀检测证实miR-126-5p可富集于MALAT1序列中。双荧光素酶报告基因检测证实MALAT1对miR-126-5p有靶向抑制作用。下调miR-126-5p可抵消MALAT1下调对高糖诱导的RECs的影响。
结论
MALAT1通过抑制miR126-5p并诱导RECs增殖、迁移和血管生成来促进PDR。
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