Department of Orthopedics, Dongzhimen Hospital Beijing University of Chinese Medicine, 116 Cuiping West Road, Tongzhou District, Beijing, 101121, China.
Inflammation. 2024 Aug;47(4):1313-1327. doi: 10.1007/s10753-024-01978-8. Epub 2024 Feb 10.
Spinal cord injury (SCI) is a serious, prolonged, and irreversible injury with few therapeutic options. Albiflorin (AF) possesses powerful pharmacodynamic properties and exerts protective effects against neuroinflammation. However, no research has examined the neuroprotective effect of AF following SCI. Rats were received laminectomy to establish SCI animal model and treated with AF (20 mg/kg and 40 mg/kg). Behavioral experiments were conducted to assess the impacts of AF on motor function after SCI in rats. Hematoxylin-eosin (HE) staining, Nissl staining, and Prussian Blue staining were performed to observe histological changes, neuronal damage, and iron deposition, respectively. Transmission electron microscope was adopted to observe the ultrastructure of spinal cord tissues. Immunofluorescence assay was performed to examine neurons and microglia. ELISA assay was used to examine the production of cytokines. Western blot assay was used to detect the expression level of ferroptosis-related proteins. Microglia BV-2 cells were induced by LPS to mimic the neuroinflammatory condition. Cell viability was assessed by CCK-8 assay, and lipid peroxidase level was measured by C11 BODIPY 581/591 staining. Molecular docking technology was utilized to confirm the relationship between AF and LSD1. AF improved the motor functional recovery after SCI in rats. Meanwhile, AF attenuated neuron apoptosis and microglia activation, reduced the production of pro-inflammatory cytokines and iron accumulation, and inhibited spinal cord ferroptosis following SCI in rats. LSD1 was verified to be a target protein of AF, and AF could concentration-dependently downregulate LSD1 expression in injured spinal cords in vivo and LPS-induced BV-2 cells in vitro. In addition, AF not only inhibited ferroptosis through reducing lipid peroxidase and iron levels and regulating ferroptosis-related proteins, but also inhibited microglial activation and reduced pro-inflammatory cytokines production in LPS-induced BV-2 cells; however, these changes were partly counteracted by LSD1 overexpression. AF could reduce microglial activation and ferroptosis, attenuate neuroinflammation, and improve functional recovery following SCI by downregulating LSD1, providing novel therapeutic strategies for the treatment of SCI.
脊髓损伤 (SCI) 是一种严重、持久且不可逆转的损伤，治疗选择有限。白芍总苷 (AF) 具有强大的药效学特性，并对神经炎症具有保护作用。然而，目前尚无研究探讨 AF 对 SCI 后的神经保护作用。
大鼠接受椎板切除术以建立 SCI 动物模型，并接受 AF（20mg/kg 和 40mg/kg）治疗。行为实验用于评估 AF 对 SCI 后大鼠运动功能的影响。苏木精-伊红 (HE) 染色、尼氏染色和普鲁士蓝染色分别用于观察组织学变化、神经元损伤和铁沉积。采用透射电子显微镜观察脊髓组织的超微结构。免疫荧光检测神经元和小胶质细胞。ELISA 检测细胞因子的产生。Western blot 检测铁死亡相关蛋白的表达水平。用 LPS 诱导 BV-2 细胞模拟神经炎症状态，通过 CCK-8 检测细胞活力，通过 C11 BODIPY 581/591 染色检测脂质过氧化水平。利用分子对接技术证实 AF 与 LSD1 的关系。
AF 改善了 SCI 后大鼠的运动功能恢复。同时，AF 减弱了神经元凋亡和小胶质细胞激活，减少了促炎细胞因子的产生和铁积累，并抑制了 SCI 后大鼠脊髓铁死亡。LSD1 被验证为 AF 的靶蛋白，AF 可以在体内 SCI 损伤脊髓和体外 LPS 诱导的 BV-2 细胞中浓度依赖性地下调 LSD1 表达。此外，AF 通过降低脂质过氧化和铁水平以及调节铁死亡相关蛋白来抑制铁死亡，同时抑制 LPS 诱导的 BV-2 细胞中小胶质细胞的激活和减少促炎细胞因子的产生；然而，这些变化在 LSD1 过表达时部分被抵消。
AF 通过下调 LSD1 减少小胶质细胞激活和铁死亡，减轻神经炎症，并改善 SCI 后的功能恢复，为 SCI 的治疗提供了新的治疗策略。
