Department of Endocrinology, The Central Hospital of Wuhan, Wuhan, Hubei, China.
Eur Rev Med Pharmacol Sci. 2020 Feb;24(3):1325-1337. doi: 10.26355/eurrev_202002_20190.
LncRNA nuclear enriched abundant transcript 1 (NEAT1) has been reported to play an oncogenic role in the occurrence and development of diabetic nephropathy (DN). The aim of our study was to investigate the potential mechanism by which NEAT1 facilitates the progression of DN.
Quantitative Real-time polymerase chain reaction (qRT-PCR) was carried out to determine the abundance of NEAT1, kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), proliferating cell nuclear antigen (PCNA), Cyclin D1, P38, apoptosis signal-regulating kinase 1 (ASK1), Fibronectin, α smooth muscle actin (α-SMA) and miR-23c in the serum of DN patients, normal patients and mouse mesangial cells (MMCs). Cell proliferation was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), qRT-PCR and Western blot assays. Flow cytometry and Western blot were applied to measure apoptosis of MMCs. Cell fibrosis and epithelial-to-mesenchymal transition (EMT) were analyzed by qRT-PCR and Western blot. The binding sites between miR-23c and NEAT1 were predicted by starBase bioinformatics software, and the relationship was verified by dual-luciferase reporter assay.
The enrichment of NEAT1 was elevated in the serum of DN patients and MMCs induced by high concentration of glucose. NEAT1 overexpression accelerated proliferation, fibrosis and EMT and restrained apoptosis of MMCs induced by high concentration of glucose. MiR-23c bound to NEAT1, and the inhibition of miR-23c counteracted the suppressive effect of NEAT1 depletion on proliferation, fibrosis and EMT of MMCs induced by high concentration glucose.
LncRNA NEAT1 promoted proliferation, fibrosis and EMT while impeded apoptosis of MMCs through sponging miR-23c. LncRNA NEAT1 and miR-23c might be underlying therapeutic targets for the treatment of DN.
长链非编码 RNA 核富集丰富转录本 1（NEAT1）已被报道在糖尿病肾病（DN）的发生和发展中发挥致癌作用。本研究旨在探讨 NEAT1 促进 DN 进展的潜在机制。
采用实时定量聚合酶链反应（qRT-PCR）测定 DN 患者、正常患者和小鼠肾小球系膜细胞（MMC）血清中 NEAT1、肾损伤分子-1（KIM-1）、中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、增殖细胞核抗原（PCNA）、细胞周期蛋白 D1、P38、凋亡信号调节激酶 1（ASK1）、纤维连接蛋白、α平滑肌肌动蛋白（α-SMA）和 miR-23c 的丰度。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、qRT-PCR 和 Western blot 测定细胞增殖。流式细胞术和 Western blot 用于测定 MMC 凋亡。通过 qRT-PCR 和 Western blot 分析细胞纤维化和上皮间质转化（EMT）。通过 starBase 生物信息学软件预测 miR-23c 与 NEAT1 的结合位点，并通过双荧光素酶报告基因实验验证。
DN 患者和高浓度葡萄糖诱导的 MMC 血清中 NEAT1 的丰度升高。NEAT1 过表达加速了高浓度葡萄糖诱导的 MMC 增殖、纤维化和 EMT，并抑制了凋亡。miR-23c 与 NEAT1 结合，抑制 miR-23c 可逆转 NEAT1 耗竭对高浓度葡萄糖诱导的 MMC 增殖、纤维化和 EMT 的抑制作用。
长链非编码 RNA NEAT1 通过海绵 miR-23c 促进 MMC 增殖、纤维化和 EMT，同时抑制其凋亡。LncRNA NEAT1 和 miR-23c 可能是治疗糖尿病肾病的潜在治疗靶点。
