Xu Meimei, Yuan Shan, Luo Xing, Xu Mengsi, Hu Guangze, He Zhe, Yang Xinyuan, Gao Rui
Department of Biochemistry, College of Medicine, Shihezi University, Shihezi, Xinjiang, China.
State Key Laboratory of Sheep Genetic Improvement and Healthy Production, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi, Xinjiang, China.
PLoS One. 2025 Jan 23;20(1):e0317710. doi: 10.1371/journal.pone.0317710. eCollection 2025.
Long non-coding RNAs (lncRNAs) are among the most abundant types of non-coding RNAs in the genome and exhibit particularly high expression levels in the brain, where they play crucial roles in various neurophysiological and neuropathological processes. Although ischemic stroke is a complex multifactorial disease, the involvement of brain-derived lncRNAs in its intricate regulatory networks remains inadequately understood. In this study, we established a cerebral ischemia-reperfusion injury model using middle cerebral artery occlusion (MCAO) in male Sprague-Dawley rats. High-throughput sequencing was performed to profile the expression of cortical lncRNAs post-stroke, with subsequent validation using RT-PCR and qRT-PCR. Among the 31,183 lncRNAs detected in the rat cerebral cortex, 551 were differentially expressed between the MCAO and sham-operated groups in the ipsilateral cortex (fold change ≥2.0, P < 0.05). An integrated analysis of the 20 most abundant and significantly differentially expressed lncRNAs (DELs) identified 25 core cytoplasmic DELs, which were used to construct an interaction network based on their targeting relationships. This led to the establishment of a comprehensive lncRNA-miRNA-mRNA regulatory network comprising 12 lncRNAs, 16 sponge miRNAs, and 191 target mRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed that differentially expressed mRNAs (DEmRNAs) were significantly enriched in stroke-related pathways. Our analysis predicted four key lncRNAs, four miRNAs, and eleven crucial mRNAs involved in post-transcriptional regulation through competing endogenous RNA (ceRNA) mechanisms. These molecules were shown to participate extensively in post-stroke processes, including angiogenesis, axonal regeneration, inflammatory responses, microglial activation, blood-brain barrier (BBB) disruption, apoptosis, autophagy, ferroptosis, and thrombocytopenia. These findings highlight the role of lncRNAs as multi-level regulators in the complex network of post-stroke mechanisms, providing novel insights into the pathophysiological processes of stroke.
长链非编码RNA(lncRNAs)是基因组中最丰富的非编码RNA类型之一,在大脑中表现出特别高的表达水平,它们在各种神经生理和神经病理过程中发挥着关键作用。尽管缺血性中风是一种复杂的多因素疾病,但脑源性lncRNAs在其复杂调控网络中的参与情况仍未得到充分了解。在本研究中,我们使用雄性Sprague-Dawley大鼠大脑中动脉闭塞(MCAO)建立了脑缺血再灌注损伤模型。进行高通量测序以分析中风后皮质lncRNAs的表达,随后使用RT-PCR和qRT-PCR进行验证。在大鼠大脑皮质中检测到的31,183个lncRNAs中,551个在MCAO组和假手术组的同侧皮质中差异表达(倍数变化≥2.0,P<0.05)。对20个最丰富且差异显著的lncRNAs(DELs)进行综合分析,鉴定出25个核心细胞质DELs,并根据它们的靶向关系构建了一个相互作用网络。这导致建立了一个由12个lncRNAs、16个海绵miRNAs和191个靶mRNA组成的综合lncRNA-miRNA-mRNA调控网络。基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析表明,差异表达的mRNA(DEmRNAs)在中风相关通路中显著富集。我们的分析预测了四个关键lncRNAs、四个miRNAs和十一个关键mRNA通过竞争性内源RNA(ceRNA)机制参与转录后调控。这些分子被证明广泛参与中风后的过程,包括血管生成、轴突再生、炎症反应、小胶质细胞激活、血脑屏障(BBB)破坏、细胞凋亡、自噬、铁死亡和血小板减少。这些发现突出了lncRNAs作为中风后复杂机制网络中多层次调节因子的作用,为中风的病理生理过程提供了新的见解。