Neurosurgery Unit, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Intensive Care Unit, The First People's Hospital of Suqian City, Suqian, China.
Cell Transplant. 2024 Jan-Dec;33:9636897241233040. doi: 10.1177/09636897241233040.
Spinal cord injury (SCI) severely affects the quality of life and autonomy of patients, and effective treatments are currently lacking. Autophagy, an essential cellular metabolic process, plays a crucial role in neuroprotection and repair after SCI. Glycoprotein non-metastatic melanoma protein B (GPNMB) has been shown to promote neural regeneration and synapse reconstruction, potentially through the facilitation of autophagy. However, the specific role of GPNMB in autophagy after SCI is still unclear. In this study, we utilized the spinal cord transection method to establish SCI rats model and overexpressed GPNMB using adenoviral vectors. We assessed tissue damage using hematoxylin and eosin (H&E) and Nissl staining, and observed cell apoptosis using TUNEL staining. We evaluated the inflammatory response by measuring inflammatory factors using enzyme-linked immunosorbent assay (ELISA). In addition, we measured reactive oxygen species (ROS) levels using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), and assessed oxidative stress levels by measuring malondialdehyde (MDA) and glutathione (GSH) using ELISA. To evaluate autophagy levels, we performed immunofluorescence staining for the autophagy marker Beclin-1 and conducted Western blot analysis for autophagy-related proteins. We also assessed limb recovery through functional evaluation. Meanwhile, we induced cell injury using lipopolysaccharide (LPS) and added an autophagy inhibitor to verify the impact of GPNMB on SCI through autophagy modulation. The results demonstrated that GPNMB alleviated the inflammatory response, reduced oxidative stress levels, inhibited cell apoptosis, and promoted autophagy following SCI. Inhibiting autophagy reversed the effects of GPNMB. These findings suggest that GPNMB promotes neural injury repair after SCI, potentially through attenuating the inflammatory response, reducing oxidative stress, and inhibiting cell apoptosis.
脊髓损伤 (SCI) 严重影响患者的生活质量和自主性,目前缺乏有效的治疗方法。自噬是一种重要的细胞代谢过程,在 SCI 后神经保护和修复中起着至关重要的作用。糖蛋白非转移性黑色素瘤蛋白 B (GPNMB) 已被证明能促进神经再生和突触重建,可能是通过促进自噬来实现的。然而,GPNMB 在 SCI 后自噬中的具体作用尚不清楚。在这项研究中,我们利用脊髓横断方法建立了 SCI 大鼠模型,并利用腺病毒载体过表达 GPNMB。我们使用苏木精和伊红 (H&E) 和尼氏染色评估组织损伤,使用 TUNEL 染色观察细胞凋亡。我们通过酶联免疫吸附试验 (ELISA) 测量炎症因子来评估炎症反应。此外,我们使用 2',7'-二氯二氢荧光素二乙酸酯 (DCFH-DA) 测量活性氧 (ROS) 水平,并通过 ELISA 测量丙二醛 (MDA) 和谷胱甘肽 (GSH) 来评估氧化应激水平。为了评估自噬水平,我们进行了自噬标志物 Beclin-1 的免疫荧光染色,并进行了自噬相关蛋白的 Western blot 分析。我们还通过功能评估来评估肢体恢复情况。同时,我们使用脂多糖 (LPS) 诱导细胞损伤,并添加自噬抑制剂来验证 GPNMB 通过自噬调节对 SCI 的影响。结果表明,GPNMB 减轻了 SCI 后的炎症反应,降低了氧化应激水平,抑制了细胞凋亡,并促进了自噬。抑制自噬逆转了 GPNMB 的作用。这些发现表明,GPNMB 通过减轻炎症反应、降低氧化应激和抑制细胞凋亡来促进 SCI 后的神经损伤修复。