LncRNA NEAT1 aggravates human microvascular endothelial cell injury by inhibiting the Apelin/Nrf2/HO-1 signalling pathway in type 2 diabetes mellitus with obstructive sleep apnoea.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Epigenetics Pub Date : 2024-12-01 Epub Date: 2024-01-17 DOI:10.1080/15592294.2023.2293409
Kai Chen, Baiqing Ou, Quan Huang, Daqing Deng, Yi Xiang, Fang Hu
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引用次数: 0

Abstract

Long noncoding RNAs (lncRNAs) regulate the progression of type 2 diabetes mellitus complicated with obstructive sleep apnoea (T2DM-OSA). However, the role of the lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) in T2DM-OSA remains unknown. This study aimed to reveal the function of NEAT1 in T2DM-OSA and the underlying mechanism. KKAy mice were exposed to intermittent hypoxia (IH) or intermittent normoxia to generate a T2DM-OSA mouse model. HMEC-1 cells were treated with high glucose (HG) and IH to construct a T2DM-OSA cell model. RNA expression was detected by qRT-PCR. The protein expression of Apelin, NF-E2-related factor 2 (Nrf2), haem oxygenase-1 (HO-1), and up-frameshift suppressor 1 (UPF1) was assessed using western blot. Cell injury was evaluated using flow cytometry, enzyme-linked immunosorbent assay, and oxidative stress kit assays. RIP, RNA pull-down, and actinomycin D assays were performed to determine the associations between NEAT1, UPF1, and Apelin. NEAT1 expression was upregulated in the aortic vascular tissues of mice with T2DM exposed to IH and HMEC-1 cells stimulated with HG and IH, whereas Apelin expression was downregulated. The absence of NEAT1 protected HMEC-1 cells from HG- and IH-induced damage. Furthermore, NEAT1 destabilized Apelin mRNA by recruiting UPF1. Apelin overexpression decreased HG- and IH-induced injury to HMEC-1 cells by activating the Nrf2/HO-1 pathway. Moreover, NEAT1 knockdown reduced HG- and IH-induced injury to HMEC-1 cells through Apelin. NEAT1 silencing reduced HMEC-1 cell injury through the Apelin/Nrf2/HO-1 signalling pathway in T2DM-OSA.Abbreviations: LncRNAs, long non-coding RNAs; T2DM, type 2 diabetes mellitus; OSA, obstructive sleep apnoea; NEAT1, nuclear paraspeckle assembly transcript 1; IH, intermittent hypoxia; HMEC-1, human microvascular endothelial cells; HG, high glucose; Nrf2, NF-E2-related factor 2; UPF1, up-frameshift suppressor 1; HO-1, haem oxygenase-1; qRT-PCR, quantitative real-time polymerase chain reaction; ELISA, enzyme-linked immunosorbent assay; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TNF-α, tumour necrosis factor-α; CCK-8, Cell Counting Kit-8; IL-1β, interleukin-1β; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase; RIP, RNA immunoprecipitation; SD, standard deviations; GSH, glutathione; AIS, acute ischaemic stroke; HMGB1, high mobility group box-1 protein; TLR4, toll-like receptor 4.

LncRNA NEAT1 通过抑制阻塞性睡眠呼吸暂停的 2 型糖尿病患者的 Apelin/Nrf2/HO-1 信号通路,加重人体微血管内皮细胞损伤。
长非编码 RNA(lncRNA)可调控 2 型糖尿病并发阻塞性睡眠呼吸暂停(T2DM-OSA)的进展。然而,lncRNA核旁斑块组装转录本1(NEAT1)在T2DM-OSA中的作用仍然未知。本研究旨在揭示 NEAT1 在 T2DM-OSA 中的功能及其内在机制。将 KKAy 小鼠暴露于间歇性低氧(IH)或间歇性常氧,以产生 T2DM-OSA 小鼠模型。用高糖(HG)和IH处理HMEC-1细胞,构建T2DM-OSA细胞模型。通过 qRT-PCR 检测 RNA 表达。用 Western 印迹法评估了 Apelin、NF-E2 相关因子 2(Nrf2)、血氧合酶-1(HO-1)和上帧移位抑制因子 1(UPF1)的蛋白表达。细胞损伤采用流式细胞术、酶联免疫吸附测定法和氧化应激试剂盒测定法进行评估。为了确定 NEAT1、UPF1 和 Apelin 之间的关联,还进行了 RIP、RNA 拉取和放线菌素 D 检测。在暴露于 IH 的 T2DM 小鼠主动脉血管组织和受到 HG 和 IH 刺激的 HMEC-1 细胞中,NEAT1 表达上调,而 Apelin 表达下调。NEAT1 的缺失可保护 HMEC-1 细胞免受 HG 和 IH 诱导的损伤。此外,NEAT1 通过招募 UPF1 使 Apelin mRNA 失稳。Apelin 的过表达可通过激活 Nrf2/HO-1 通路减少 HG 和 IH 诱导的 HMEC-1 细胞损伤。此外,敲除 NEAT1 可通过 Apelin 减少 HG 和 IH 诱导的对 HMEC-1 细胞的损伤。在T2DM-OSA中,沉默NEAT1可通过Apelin/Nrf2/HO-1信号通路减少HMEC-1细胞损伤:缩写:LncRNAs,长非编码 RNAs;T2DM,2 型糖尿病;OSA,阻塞性睡眠呼吸暂停;NEAT1,核旁斑块组装转录本 1;IH,间歇性缺氧;HMEC-1,人微血管内皮细胞;HG,高血糖;Nrf2,NF-E2 相关因子 2;UPF1,上移抑制因子 1;HO-1,血氧合酶-1;qRT-PCR,定量实时聚合酶链反应;ELISA,酶联免疫吸附试验;GAPDH,3-磷酸甘油醛脱氢酶;TNF-α,肿瘤坏死因子-α;CCK-8,细胞计数试剂盒-8;IL-1β,白细胞介素-1β;ROS,活性氧;MDA,丙二醛;SOD:超氧化物歧化酶;RIP:RNA 免疫沉淀;SD:标准差;GSH:谷胱甘肽;AIS:急性缺血性中风;HMGB1:高迁移率组盒-1 蛋白;TLR4:收费样受体 4。
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来源期刊
Epigenetics
Epigenetics 生物-生化与分子生物学
CiteScore
6.80
自引率
2.70%
发文量
82
审稿时长
3-8 weeks
期刊介绍: Epigenetics publishes peer-reviewed original research and review articles that provide an unprecedented forum where epigenetic mechanisms and their role in diverse biological processes can be revealed, shared, and discussed. Epigenetics research studies heritable changes in gene expression caused by mechanisms others than the modification of the DNA sequence. Epigenetics therefore plays critical roles in a variety of biological systems, diseases, and disciplines. Topics of interest include (but are not limited to): DNA methylation Nucleosome positioning and modification Gene silencing Imprinting Nuclear reprogramming Chromatin remodeling Non-coding RNA Non-histone chromosomal elements Dosage compensation Nuclear organization Epigenetic therapy and diagnostics Nutrition and environmental epigenetics Cancer epigenetics Neuroepigenetics
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