Zhu Wenwen, Yang Xingchun, Li Nanxi, Zhang Bin, Huang Lishan, Cheng Hanxing, Wu Xiao, Zhang Dechou, Li Sen, Xu Houping
Geriatric Department, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou, Sichuan, China.
Department of Acupuncture and Tuina, Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University, Luzhou, Sichuan, China.
Medicine (Baltimore). 2025 Jan 17;104(3):e40965. doi: 10.1097/MD.0000000000040965.
Insomnia is increasingly common and poses significant health risks. The aims of this study are to identify apoptosis-related genes and potential biomarkers for insomnia and to find new therapeutic targets. Insomnia gene expression profiles were downloaded from the Gene Expression Omnibus database, and differentially expressed genes in normal and insomnia samples were identified by limma rapid differential analysis, and then the major modular genes with clinical relevance to insomnia were analyzed using the Weighted Gene Co-Expression Network Analysis, and intersections were obtained with the differentially expressed genes as well as with apoptotic gene databases. We validated apoptosis-related differentially expressed genes, enriched and analyzed the specific biological process of insomnia and related signaling pathways. In addition, we constructed a protein-protein interaction network and obtained Top10 hub genes using Cytoscape. We selected 3 of them as hub genes and compared their expression in normal hippocampal neuronal cells and hippocampal neuronal cells of the model group exposed to corticosterone induction by Western Blot and qRT-PCR experiments. A total of 190 differentially expressed apoptosis-related genes were identified in insomnia, and BCL2, SOCS3, and IL7R were identified as important hub genes. Enrichment analysis showed that the occurrence of apoptosis in insomnia was mainly related to "PI3K-Akt signaling pathway," "JAK-STAT signaling pathway," "P53 signaling pathway" and so on. GO analysis showed that apoptosis in insomnia was mainly related to "immune response," "T cell differentiation in thymus," and "positive regulation of MAPK cascade." Western Blot and qRT-PCR experiments showed that BCL2, SOCS3, IL7R antiapoptotic indexes were under-expressed in modeled hippocampal neuronal cells compared to normal hippocampal neuronal cells. This study emphasizes the role of apoptosis-related genes in insomnia and preliminarily predicts that the occurrence of insomnia is closely related to apoptosis. Compared to the normal group, the antiapoptotic ability of hippocampal neurons in the model group is reduced. Although BCL2 has been studied in the context of sleep deprivation, SOCS3 and IL7R have not yet been explored in insomnia. Insomnia and sleep deprivation involve similar pathways, but due to different mechanisms and types of insomnia, gene expression may vary.
失眠日益普遍，并带来重大健康风险。本研究的目的是识别与失眠相关的凋亡基因和潜在生物标志物，并寻找新的治疗靶点。从基因表达综合数据库下载失眠基因表达谱，通过limma快速差异分析识别正常样本和失眠样本中的差异表达基因，然后使用加权基因共表达网络分析来分析与失眠具有临床相关性的主要模块基因，并与差异表达基因以及凋亡基因数据库进行交集分析。我们验证了与凋亡相关的差异表达基因，对失眠的特定生物学过程和相关信号通路进行了富集和分析。此外，我们构建了蛋白质-蛋白质相互作用网络，并使用Cytoscape获得了前10个枢纽基因。我们从中选择3个作为枢纽基因，并通过蛋白质免疫印迹法和实时定量聚合酶链反应实验比较它们在正常海马神经元细胞和暴露于皮质酮诱导的模型组海马神经元细胞中的表达。在失眠中总共鉴定出190个与凋亡相关的差异表达基因，BCL2、SOCS3和IL7R被鉴定为重要的枢纽基因。富集分析表明，失眠中凋亡的发生主要与“PI3K-Akt信号通路”、“JAK-STAT信号通路”、“P53信号通路”等有关。基因本体分析表明，失眠中的凋亡主要与“免疫反应”、“胸腺中的T细胞分化”和“MAPK级联的正调控”有关。蛋白质免疫印迹法和实时定量聚合酶链反应实验表明，与正常海马神经元细胞相比，模型组海马神经元细胞中BCL2、SOCS3、IL7R的抗凋亡指标表达下调。本研究强调了凋亡相关基因在失眠中的作用，并初步预测失眠的发生与凋亡密切相关。与正常组相比，模型组海马神经元的抗凋亡能力降低。虽然BCL2已在睡眠剥夺的背景下进行了研究，但SOCS3和IL7R在失眠中尚未得到探索。失眠和睡眠剥夺涉及相似的途径，但由于失眠的机制和类型不同，基因表达可能会有所差异。
