Zhou Wu, Hei Bo, Liu Yihao, Wang Chengyun, Wang Chunliang, Ding Zihan
Department of Neurosurgery, the 1st Affiliated Hospital, Jiangxi Medical College, Nanchang University, 17 Yongwai Street, Nanchang, 330006, Jiangxi, China.
Department of Neurosurgery, Peking University People's Hospital, Peking University, No.11 Xizhimen South Street, Beijing, Beijing, 100044, China.
Mol Neurobiol. 2025 Apr 24. doi: 10.1007/s12035-025-04954-x.
Spinal cord injury (SCI) is a serious complication of spinal fractures and/or dislocations, characterized by sensory and motor dysfunction in the trunk and limbs. The pathogenesis of SCI is highly complex and remains poorly understood. The role of O-GlcNAc modification and FOXD3 in SCI was studied in this study. The cell and animal models of SCI were established by HO stimulation and heavy object impact method, respectively. HE and Nissl staining were used to analyze pathological changes and neuronal loss in the spinal cord tissues. The motor ability of rats was assessed by BBB score, ladder climbing, and grid climbing tests. Cell viability and apoptosis were assessed by CCK8, flow cytometry, and TUNEL staining, respectively. Co-IP assay detected O-GlcNAc modification level of FOXD3 protein. The interaction between FOXD3 and STUB1 promoter was analyzed by dual luciferase reporter gene and ChIP assays. O-GlcNAc modification level was significantly elevated in the cell and animal models of SCI. O-GlcNAc modification increased both the protein stability and expression of FOXD3. O-GlcNAc modification inhibition or FOXD3 knockdown reduced oxidative stress damage and apoptosis in HO-treated PC12 cells. Moreover, FOXD3 mediated transcriptional inhibition of STUB1, and STUB1 induced HMGB1 ubiquitination and degradation in PC12 cells. STUB1 knockdown or HMGB1 overexpression negated the protective effects of FOXD3 knockdown on HO-mediated oxidative stress damage and apoptosis in PC12 cells. Inhibiting the O-GlcNAc modification of FOXD3 alleviated oxidative stress damage and apoptosis in nerve cells to mitigate SCI by enhancing STUB1-induced HMGB1 ubiquitination degradation.
脊髓损伤（SCI）是脊柱骨折和/或脱位的一种严重并发症，其特征为躯干和四肢的感觉及运动功能障碍。SCI的发病机制高度复杂，目前仍了解甚少。本研究探讨了O-GlcNAc修饰和FOXD3在SCI中的作用。分别采用HO刺激法和重物撞击法建立SCI的细胞和动物模型。运用苏木精-伊红（HE）染色和尼氏染色分析脊髓组织的病理变化及神经元丢失情况。通过BBB评分、爬梯试验和网格攀爬试验评估大鼠的运动能力。分别采用CCK8法、流式细胞术和TUNEL染色检测细胞活力和凋亡情况。免疫共沉淀（Co-IP）试验检测FOXD3蛋白的O-GlcNAc修饰水平。采用双荧光素酶报告基因试验和染色质免疫沉淀（ChIP）试验分析FOXD3与STUB1启动子之间的相互作用。在SCI的细胞和动物模型中，O-GlcNAc修饰水平显著升高。O-GlcNAc修饰增加了FOXD3的蛋白稳定性和表达。抑制O-GlcNAc修饰或敲低FOXD3可减轻HO处理的PC12细胞中的氧化应激损伤和凋亡。此外，FOXD3介导对STUB1的转录抑制，并且STUB1在PC12细胞中诱导HMGB1的泛素化和降解。敲低STUB1或过表达HMGB1可消除FOXD3敲低对HO介导的PC12细胞氧化应激损伤和凋亡的保护作用。抑制FOXD3的O-GlcNAc修饰可通过增强STUB1诱导的HMGB1泛素化降解减轻神经细胞中的氧化应激损伤和凋亡，从而减轻脊髓损伤。
