Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, China; Department of Neurology, The Affiliated Hospital of Chengde Medical University, Chengde 067000, China; Hebei Key Laboratory of Panvascular Diseases, The Affiliated Hospital of Chengde Medical University, Chengde, China.
Department of Neurology, The Affiliated Hospital of Chengde Medical University, Chengde 067000, China.
Neuroscience. 2024 Nov 12;560:130-142. doi: 10.1016/j.neuroscience.2024.09.038. Epub 2024 Sep 19.
Over the years, the neuroprotective potential of bone marrow mesenchymal stem cells (BMSCs) in acute ischemic stroke has attracted significant attention. However, BMSCs face challenges like short metabolic cycles and low survival rates post-transplant. Polypyrimidine tract-binding protein 1 (PTBP1) is an immunomodulatory RNA-binding protein that regulates the cell cycle and increases cell viability. This study investigated the protective effects and underlying mechanism of PTBP1 knockdown in BMSCs (PTBP1KD-BMSCs) following ischemia-reperfusion injury (IRI) in neurons. BMSCs were isolated from Sprague-Dawley rat femurs and characterized through flow cytometry and differentiation induction. PTBP1 knockdown inhibited BMSCs proliferation. Co-culture with PTBP1KD-BMSCs decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels, while increasing glutathione (GSH) production in oxygen and glucose deprivation/reperfusion-induced PC12 cells. Transcriptome sequencing analysis of PC12 cells suggested that the protective effect of PTBP1KD-BMSCs against injury may involve ferroptosis. Furthermore, western blotting showed upregulation of glutathione synthetase (GSS), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) in PTBP1KD-BMSCs, known negative regulators of ferroptosis. Moreover, PTBP1KD-BMSCs inhibited p38MAPK and JNK activation. In addition, PTBP1KD-BMSCs transplantation into middle cerebral artery occlusion/reperfusion (MCAO/R) rats reduced cerebral infarction volume and improved neurological function. Immunofluorescence analysis confirmed the upregulation of GSS expression in neurons of the ischemic cortex, while immunohistochemistry indicated a downregulation of p-P38. These result suggest that PTBP1KD-BMSCs can alleviate neuronal IRI by reducing oxidative stress, inhibiting ferroptosis, and modulating the MAPK pathway, providing a theoretical basis for potential treatment strategies for cerebral IRI.
多年来，骨髓间充质干细胞（BMSCs）在急性缺血性脑卒中的神经保护潜力引起了广泛关注。然而，BMSCs 在移植后存在代谢周期短和存活率低等挑战。聚嘧啶 tract-binding 蛋白 1（PTBP1）是一种免疫调节 RNA 结合蛋白，可调节细胞周期并提高细胞活力。本研究探讨了缺血再灌注损伤（IRI）后 BMSCs 中 PTBP1 敲低（PTBP1KD-BMSCs）对神经元的保护作用及其潜在机制。BMSCs 从 Sprague-Dawley 大鼠股骨中分离出来，并通过流式细胞术和分化诱导进行鉴定。PTBP1 敲低抑制 BMSCs 的增殖。与 PTBP1KD-BMSCs 共培养可降低氧葡萄糖剥夺/再灌注诱导的 PC12 细胞中活性氧（ROS）和丙二醛（MDA）的水平，同时增加谷胱甘肽（GSH）的产生。PC12 细胞的转录组测序分析表明，PTBP1KD-BMSCs 对损伤的保护作用可能涉及铁死亡。此外，Western blot 显示，PTBP1KD-BMSCs 中谷胱甘肽合成酶（GSS）、谷胱甘肽过氧化物酶 4（GPX4）和溶质载体家族 7 成员 11（SLC7A11）的表达上调，这些是铁死亡的负调节因子。此外，PTBP1KD-BMSCs 抑制了 p38MAPK 和 JNK 的激活。此外，PTBP1KD-BMSCs 移植到大脑中动脉闭塞/再灌注（MCAO/R）大鼠中可减少脑梗死体积并改善神经功能。免疫荧光分析证实缺血皮质神经元中 GSS 表达上调，免疫组织化学分析表明 p-P38 下调。这些结果表明，PTBP1KD-BMSCs 可通过减轻氧化应激、抑制铁死亡和调节 MAPK 通路来减轻神经元 IRI，为脑 IRI 的潜在治疗策略提供了理论依据。
