杜鹃素通过JAK2/STAT3通路调节巨噬细胞/小胶质细胞极化，从而在脊髓损伤中发挥神经保护作用。

Farrerol confers neuroprotection in spinal cord injury by regulating macrophages/microglia polarization through the JAK2/STAT3 pathway.

作者信息

Chen Yue, Ren Lü, Xia Jinzhi, Li Bohan, Yang Yi, Li Jing, Tao Lu, Song Xue, Lü Hezuo, Hu Jianguo

机构信息

Department of Rehabilitation, First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China; Department of Laboratory Medicine, Bengbu Medical University, Bengbu, 233000, China; Anhui Province Key Laboratory of Basic and Translational Research of Inflammation-related Diseases, Bengbu 233000, China.

Department of Laboratory Medicine, Bengbu Medical University, Bengbu, 233000, China; Department of Clinical Laboratory, First Affiliated Hospital of Bengbu Medical University, Bengbu 233000, China; Anhui Province Key Laboratory of Basic and Translational Research of Inflammation-related Diseases, Bengbu 233000, China.

出版信息

Exp Neurol. 2025 Aug 25;394:115448. doi: 10.1016/j.expneurol.2025.115448.

DOI:10.1016/j.expneurol.2025.115448

PMID:40865787

Abstract

BACKGROUND AND AIMS

Spinal cord injury (SCI) causes secondary damage characterized by neuroinflammation and imbalanced macrophages/microglia polarization, worsening neuronal loss and functional decline. Farrerol (FAR), a natural flavonoid with anti-inflammatory properties, has not been studied for SCI treatment. This work assesses FAR's neuroprotection through macrophages/microglia polarization regulation and explores its mechanisms.

METHODS

C57BL/6 mice with spinal cord injury were randomly assigned to three groups: Sham, SCI, and SCI + FAR. Motor function was evaluated using locomotor scoring, while lesion size, myelin integrity, and neuronal apoptosis were assessed via histology, immunofluorescence, and Western blot. Spinal inflammatory cytokine, macrophages/microglia activation, and polarization were analyzed by qRT-PCR, ELISA, immunofluorescence, and flow cytometry. LPS-stimulated BV2 microglia and BV2-HT22 co-cultures evaluated FAR's effects on cytokine secretion, macrophages/microglia phenotypes, and neuronal survival. Signaling mechanisms were further examined via Western blot and immunofluorescence.

RESULTS

FAR treatment significantly enhanced motor recovery in SCI mice, evidenced by elevated Basso Mouse Scale (BMS) scores, increased inclined plane angles, improved swimming performance, and refined gait patterns. It reduced lesion area, preserved myelin integrity, and attenuated neuronal apoptosis. FAR downregulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), suppressed macrophages/microglia hyperactivation while upregulating IL-10, and shifted M1/M2 polarization toward neuroprotective M2 phenotypes. In LPS-stimulated BV2 microglia, FAR attenuated inflammatory responses, inhibited M1 markers, enhanced M2 markers, and rescued HT22 neuronal apoptosis in co-cultures. These therapeutic effects may be mediated through suppression of JAK2/STAT3 phosphorylation.

CONCLUSIONS

FAR promotes functional recovery after spinal cord injury by modulating macrophages/microglia M1/M2 polarization through JAK2/STAT3 pathway inhibition, thereby attenuating neuroinflammation and neuronal death. These findings provide novel evidence supporting targeted immunomodulation for SCI treatment.

摘要

背景与目的

脊髓损伤（SCI）会导致以神经炎症和巨噬细胞/小胶质细胞极化失衡为特征的继发性损伤，从而加剧神经元丢失和功能衰退。法尔瑞林（FAR）是一种具有抗炎特性的天然黄酮类化合物，尚未针对脊髓损伤治疗进行研究。本研究通过调节巨噬细胞/小胶质细胞极化来评估FAR的神经保护作用，并探索其作用机制。

方法

将脊髓损伤的C57BL/6小鼠随机分为三组：假手术组、脊髓损伤组和脊髓损伤+FAR组。使用运动评分评估运动功能，通过组织学、免疫荧光和蛋白质印迹法评估损伤大小、髓鞘完整性和神经元凋亡。通过qRT-PCR、ELISA、免疫荧光和流式细胞术分析脊髓炎性细胞因子、巨噬细胞/小胶质细胞激活和极化情况。用脂多糖刺激BV2小胶质细胞和BV2-HT22共培养物，评估FAR对细胞因子分泌、巨噬细胞/小胶质细胞表型和神经元存活的影响。通过蛋白质印迹法和免疫荧光进一步研究信号传导机制。

结果

FAR治疗显著增强了脊髓损伤小鼠的运动恢复，表现为巴索小鼠量表（BMS）评分升高、倾斜平面角度增加、游泳性能改善和步态模式优化。它减少了损伤面积，保持了髓鞘完整性，并减轻了神经元凋亡。FAR下调促炎细胞因子（TNF-α、IL-1β、IL-6），抑制巨噬细胞/小胶质细胞过度激活，同时上调IL-10，并将M1/M2极化转变为神经保护性M2表型。在脂多糖刺激的BV2小胶质细胞中，FAR减轻了炎症反应，抑制了M1标志物，增强了M2标志物，并挽救了共培养物中HT22神经元的凋亡。这些治疗效果可能是通过抑制JAK2/STAT3磷酸化介导的。