Chen Lizhu, Yan Xiaoqiong
Department of Neurology, Hubei Provincial Hospital of Integrated Traditional and Western Medicine, Jianghan District, No. 11 Lingjiaohu Road, Wuhan, 430015 China.
Cytotechnology. 2025 Feb;77(1):41. doi: 10.1007/s10616-024-00691-6. Epub 2025 Jan 10.
Alzheimer's disease (AD) is a progressive neurological condition that causes brain shrinkage and cell death. This study aimed to identify the role of the NORAD/miR-26b-5p axis in AD. StarBase was used to examine the binding sequences of miR-26b-5p to LncRNA NORAD or its target genes, which were verified by a double luciferase reporter assay. PC12 cells were processed with Aβ to construct an AD model in vitro, and LncRNA NORAD and miR-26b-5p levels in PC12 cells were identified by RT-qPCR. Cell viability and apoptosis were measured using the MTT assay and flow cytometry, respectively. LDH release and oxidative stress-related indicators (MDA, SOD, and CAT) were detected using the corresponding kits, and the levels of Bcl-2 and Bax were assessed by western blotting and RT-qPCR. Aβ distinctly decreased LncRNA NORAD and membrane metalloendopeptidase (MME) levels in PC12 cells, while miR-26b-5p was generally increased. The LncRNA NORAD can adsorb miR-26b-5p, and the target gene of miR-26b-5p is neprilysin (MME). In the Aβ induced AD model, PC12 cell activity decreased, LDH release and apoptosis increased, oxidative stress level increased, Bax expression increased, and Bcl-2 expression decreased. LncRNA NORAD plays a protective role in AD cell models by abrogating miR-26b-5p levels. Inhibition of MME expression eliminated the protective effects of the miR-26b-5p inhibitor in AD cell models. LncRNA NORAD inhibits AD progression in vitro by modulating the miR-26b-5p-MME signaling axis. The LncRNA NORAD/miR-26b-5p is expected to be a prospective therapeutic candidate for AD.
The online version contains supplementary material available at 10.1007/s10616-024-00691-6.
阿尔茨海默病（AD）是一种进行性神经疾病，会导致脑萎缩和细胞死亡。本研究旨在确定NORAD/miR-26b-5p轴在AD中的作用。利用StarBase检测miR-26b-5p与长链非编码RNA（LncRNA）NORAD或其靶基因的结合序列，并通过双荧光素酶报告基因检测进行验证。用β-淀粉样蛋白（Aβ）处理PC12细胞以在体外构建AD模型，通过逆转录定量聚合酶链反应（RT-qPCR）鉴定PC12细胞中LncRNA NORAD和miR-26b-5p的水平。分别使用MTT法和流式细胞术检测细胞活力和细胞凋亡。使用相应试剂盒检测乳酸脱氢酶（LDH）释放和氧化应激相关指标（丙二醛、超氧化物歧化酶和过氧化氢酶），并通过蛋白质免疫印迹法和RT-qPCR评估Bcl-2和Bax的水平。Aβ明显降低PC12细胞中LncRNA NORAD和膜金属内肽酶（MME）的水平，而miR-26b-5p通常升高。LncRNA NORAD可吸附miR-26b-5p，且miR-26b-5p的靶基因是中性内肽酶（MME）。在Aβ诱导的AD模型中，PC12细胞活性降低，LDH释放和细胞凋亡增加，氧化应激水平升高，Bax表达增加，Bcl-2表达降低。LncRNA NORAD通过消除miR-26b-5p水平在AD细胞模型中发挥保护作用。抑制MME表达消除了miR-26b-5p抑制剂在AD细胞模型中的保护作用。LncRNA NORAD通过调节miR-26b-5p-MME信号轴在体外抑制AD进展。LncRNA NORAD/miR-26b-5p有望成为AD的一种有前景的治疗候选物。
在线版本包含可在10.1007/s10616-024-00691-6获取的补充材料。
