Wu Haidong, Huang Jing, Li Xiaojing, Song Yali, Chen Xuxiang, Guo Yajie
Department of Emergency, The Eighth Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China.
Organogenesis. 2025 Dec;21(1):2519641. doi: 10.1080/15476278.2025.2519641. Epub 2025 Jun 16.
Neural stem cell (NSC) possess the essential properties of pluripotency and self-renewal, making them promising candidates for the treatment of neurological disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), and spinal cord injuries. While previous studies have identified the long non-coding RNAs (lncRNAs) Pnky as a regulator of NSC differentiation into neurons via RNA splicing, its role in NSC differentiation and proliferation through the Wnt/β-catenin pathway remains unclear. In this study, we investigated the mechanism by which Pnky influences the Wnt/β-catenin pathway to promote NSC differentiation into neurons. Using cck8 assays, western blot analysis, and quantitative polymerase chain reaction (qPCR), we found that Pnky knockdown significantly enhanced NSC proliferation and promoted their differentiation into neurons. Additionally, Pnky knockdown resulted in the downregulation of the neural stem cell marker Nestin and upregulation of the neuronal marker β3-Tubulin, through activation of the β-catenin signaling pathway. Conversely, inhibiting the β-catenin pathway hindered both NSC differentiation and proliferation. These findings suggest that targeting the Pnky-mediated Wnt/β-catenin pathway may offer novel strategies for the treatment, diagnosis, and drug development of central nervous system diseases.
神经干细胞(NSC)具有多能性和自我更新的基本特性,使其成为治疗诸如阿尔茨海默病(AD)、帕金森病(PD)和脊髓损伤等神经疾病的有潜力的候选者。虽然先前的研究已确定长链非编码RNA(lncRNA)Pnky是通过RNA剪接调控神经干细胞向神经元分化的因子,但其通过Wnt/β-连环蛋白途径在神经干细胞分化和增殖中的作用仍不清楚。在本研究中,我们探究了Pnky影响Wnt/β-连环蛋白途径以促进神经干细胞向神经元分化的机制。通过细胞计数试剂盒8(cck8)检测、蛋白质免疫印迹分析和定量聚合酶链反应(qPCR),我们发现敲低Pnky可显著增强神经干细胞的增殖并促进其向神经元分化。此外,敲低Pnky通过激活β-连环蛋白信号通路,导致神经干细胞标志物巢蛋白(Nestin)表达下调,神经元标志物β3微管蛋白(β3-Tubulin)表达上调。相反,抑制β-连环蛋白途径会阻碍神经干细胞的分化和增殖。这些发现表明,靶向Pnky介导的Wnt/β-连环蛋白途径可能为中枢神经系统疾病的治疗、诊断和药物开发提供新的策略。