Epidermal Neural Crest Stem Cell Conditioned Medium Enhances Spinal Cord Injury Recovery via PI3K/AKT-Mediated Neuronal Apoptosis Suppression

IF 3.7 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Neurochemical Research Pub Date : 2024-07-18 DOI:10.1007/s11064-024-04207-8

Ziqian Ma, Tao Liu, Liang Liu, Yilun Pei, Tianyi Wang, Zhijie Wang, Yun Guan, Xinwei Zhang, Yan Zhang, Xueming Chen

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Abstract

This study aimed to assess the impact of conditioned medium from epidermal neural crest stem cells (EPI-NCSCs-CM) on functional recovery following spinal cord injury (SCI), while also exploring the involvement of the PI3K-AKT signaling pathway in regulating neuronal apoptosis. EPI-NCSCs were isolated from 10-day-old Sprague-Dawley rats and cultured for 48 h to obtain EPI-NCSC-CM. SHSY-5Y cells were subjected with H₂O₂ treatment to induce apoptosis. Cell viability and survival rates were evaluated using the CCK-8 assay and calcein-AM/PI staining. SCI contusion model was established in adult Sprague-Dawley rats to assess functional recovery, utilizing the Basso, Beattie and Bresnahan (BBB) scoring system, inclined test, and footprint observation. Neurological restoration after SCI was analyzed through electrophysiological recordings. Histological analysis included hematoxylin and eosin (H&E) staining and Nissl staining to evaluate tissue organization. Apoptosis and oxidative stress levels were assessed using TUNEL staining and ROS detection methods. Additionally, western blotting was performed to examine the expression of apoptotic markers and proteins related to the PI3K/AKT signaling pathway. EPI-NCSC-CM significantly facilitated functional and histological recovery in SCI rats by inhibiting neuronal apoptosis through modulation of the PI3K/AKT pathway. Administration of EPI-NCSCs-CM alleviated H2O2-induced neurotoxicity in SHSY-5Y cells in vitro. The use of LY294002, a PI3K inhibitor, underscored the crucial role of the PI3K/AKT signaling pathway in regulating neuronal apoptosis. This study contributes to the ongoing exploration of molecular pathways involved in spinal cord injury (SCI) repair, focusing on the therapeutic potential of EPI-NCSC-CM. The research findings indicate that EPI-NCSC-CM exerts a neuroprotective effect by suppressing neuronal apoptosis through activation of the PI3K/AKT pathway in SCI rats. These results highlight the promising role of EPI-NCSC-CM as a potential treatment strategy for SCI, emphasizing the significance of the PI3K/AKT pathway in mediating its beneficial effects.

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表皮神经干细胞条件培养基通过抑制 PI3K/AKT 介导的神经元凋亡促进脊髓损伤的恢复

本研究旨在评估表皮神经嵴干细胞条件培养基（EPI-NCSCs-CM）对脊髓损伤（SCI）后功能恢复的影响，同时探索PI3K-AKT信号通路在调节神经细胞凋亡中的参与。从10天大的Sprague-Dawley大鼠体内分离出EPI-NCSCs，培养48小时后获得EPI-NCSC-CM。用 H2O2 处理 SHSY-5Y 细胞以诱导细胞凋亡。细胞活力和存活率通过 CCK-8 检测法和钙黄绿素-AM/PI 染色法进行评估。利用 Basso、Beattie 和 Bresnahan（BBB）评分系统、倾斜试验和足迹观察，在成年 Sprague-Dawley 大鼠中建立了 SCI 挫伤模型，以评估功能恢复情况。通过电生理记录分析 SCI 后的神经恢复情况。组织学分析包括苏木精和伊红（H&E）染色以及尼氏染色，以评估组织结构。采用TUNEL染色和ROS检测方法评估细胞凋亡和氧化应激水平。此外，还进行了Western印迹以检测凋亡标志物和与PI3K/AKT信号通路相关的蛋白质的表达。通过调节PI3K/AKT通路抑制神经细胞凋亡，EPI-NCSC-CM明显促进了SCI大鼠的功能和组织学恢复。服用 EPI-NCSCs-CM 可减轻 H2O2- 诱导的 SHSY-5Y 细胞体外神经毒性。PI3K抑制剂LY294002的使用强调了PI3K/AKT信号通路在调节神经细胞凋亡中的关键作用。这项研究有助于不断探索脊髓损伤（SCI）修复的分子通路，重点是 EPI-NCSC-CM 的治疗潜力。研究结果表明，EPI-NCSC-CM通过激活PI3K/AKT通路抑制SCI大鼠神经细胞凋亡，从而发挥神经保护作用。这些结果凸显了 EPI-NCSC-CM 作为一种潜在的 SCI 治疗策略的前景，强调了 PI3K/AKT 通路在介导其有益作用方面的重要性。

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来源期刊

Neurochemical Research 医学-神经科学

CiteScore

7.70

自引率

2.30%

发文量

320

审稿时长

6 months

期刊介绍： Neurochemical Research is devoted to the rapid publication of studies that use neurochemical methodology in research on nervous system structure and function. The journal publishes original reports of experimental and clinical research results, perceptive reviews of significant problem areas in the neurosciences, brief comments of a methodological or interpretive nature, and research summaries conducted by leading scientists whose works are not readily available in English.