Department of Gerontology, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Eur Rev Med Pharmacol Sci. 2020 Nov;24(21):11259-11265. doi: 10.26355/eurrev_202011_23615.
The purpose of this study was to uncover the potential role of non-coding RNA activated by DNA damage (lncRNA NORAD) in the disease progression of diabetic cardiomyopathy (DCM) and the underlying mechanism.
Cell viability, 5-Ethynyl-2'-deoxyuridine (EdU)-positive ratio and apoptotic rate in human cardiomyocyte cell line AC16 undergoing treatment of normal-level (NG) or high-level glucose (HG) were assessed at first. NORAD level in HG-induced AC16 cells at different time points was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Subsequently, cell viability, EdU-positive ratio, and apoptotic rate in HG-induced AC16 cells overexpressing NORAD were evaluated. Next, subcellular distribution of NORAD was examined, and Dual-Luciferase reporter gene assay was performed to clarify the interaction among NORAD, miRNA-150-5p, and ZEB1. At last, rescue experiments were conducted to clarify the role of NORAD/miRNA-150-5p/ZEB1 axis in influencing the proliferation and apoptosis in HG-induced AC16 cells.
Results revealed that HG treatment suppressed the proliferative ability and stimulated apoptosis in AC16 cells. Besides, NORAD was time-dependently downregulated in HG-induced AC16 cells, and it was mainly distributed in cytoplasm. In addition, the overexpression of NORAD enhanced proliferative ability, attenuated apoptosis, and increased Bcl-2/Bax ratio in HG-induced AC16 cells. Finally, NORAD/miRNA-150-5p/ZEB1 axis was verified to protect the malignant progression of DCM.
NORAD is upregulated under high-level glucose treatment. Overexpression of NORAD protects DCM development via miRNA-150-5p/ZEB1 axis.
本研究旨在揭示 DNA 损伤激活的非编码 RNA（lncRNA NORAD）在糖尿病心肌病（DCM）疾病进展中的潜在作用及其机制。
首先评估正常水平（NG）或高水平葡萄糖（HG）处理下人心肌细胞系 AC16 的细胞活力、5-乙炔基-2'-脱氧尿苷（EdU）阳性率和凋亡率。通过实时定量聚合酶链反应（qRT-PCR）检测不同时间点 HG 诱导的 AC16 细胞中 NORAD 的水平。随后，评估过表达 NORAD 的 HG 诱导的 AC16 细胞的细胞活力、EdU 阳性率和凋亡率。接下来，检查 NORAD 的亚细胞分布，并进行双荧光素酶报告基因检测以阐明 NORAD、miRNA-150-5p 和 ZEB1 之间的相互作用。最后，进行挽救实验以阐明 NORAD/miRNA-150-5p/ZEB1 轴在影响 HG 诱导的 AC16 细胞增殖和凋亡中的作用。
结果表明，HG 处理抑制了 AC16 细胞的增殖能力并刺激其凋亡。此外，HG 诱导的 AC16 细胞中 NORAD 呈时间依赖性下调，主要分布在细胞质中。此外，NORAD 的过表达增强了 HG 诱导的 AC16 细胞的增殖能力，减弱了其凋亡，并增加了 Bcl-2/Bax 比值。最后，验证了 NORAD/miRNA-150-5p/ZEB1 轴可保护 DCM 的恶性进展。
高水平葡萄糖处理下 NORAD 上调。NORAD 的过表达通过 miRNA-150-5p/ZEB1 轴保护 DCM 的发展。
