Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, Jiangsu, People's Republic of China; Department of Orthopaedic Surgery, Changzheng Hospital, Naval Medical University, Shanghai, China; Department of Spinal Surgery, Subei People's Hospital, Clinical Medical School, Yangzhou University Affiliated Hospital, Yangzhou, China.
Department of Orthopaedic Surgery, Changzheng Hospital, Naval Medical University, Shanghai, China.
Int Immunopharmacol. 2024 Mar 30;130:111738. doi: 10.1016/j.intimp.2024.111738. Epub 2024 Feb 29.
Neuroinflammation, a critical component of the secondary injury cascade post-spinal cord injury, involves the activation of pro-inflammatory cells and release of inflammatory mediators. Resolution of neuroinflammation is closely linked to cellular autophagy. This study investigates the potential of Fisetin, a natural anti-inflammatory compound, to ameliorate neuroinflammation and confer spinal cord injury protection through the regulation of autophagy in pro-inflammatory cells.
Utilizing a rat T10 spinal cord injury model with distinct treatment groups (Sham, Fisetin-treated, and Fisetin combined with autophagy inhibitor), alongside in vitro models involving lipopolysaccharide (LPS)-stimulated microglial cell activation and co-culture with neurons, we employed techniques such as transcriptomic sequencing, histological assessments (immunofluorescence staining, etc.), molecular analyses (PCR, WB, ELISA, etc.), and behavioral evaluations to discern differences in neuroinflammation, autophagy, neuronal apoptosis, and neurological function recovery.
Fisetin significantly augmented autophagic activity in injured spinal cord tissue, crucially contributing to neurological function recovery in spinal cord-injured rats. Fisetin's autophagy-dependent effects were associated with a reduction in neuronal apoptosis at the injury site. The treatment reduced the population of CD68+ and iNOS+ cells, coupled with decreased pro-inflammatory cytokines IL-6 and TNF-α levels, through autophagy-dependent pathways. Fisetin pre-treatment attenuated LPS-induced pro-inflammatory polarization of microglial cells, with this protective effect partially blocked by autophagy inhibition. Fisetin-induced autophagy in the injured spinal cord and pro-inflammatory microglial cells was associated with significant activation of AMPK and inhibition of mTOR.
Fisetin orchestrates enhanced autophagy in pro-inflammatory microglial cells through the AMPK-mTOR signaling pathway, thereby mitigating neuroinflammation and reducing the apoptotic effects of neuroinflammation on neurons. This mechanistic insight significantly contributes to the protection and recovery of neurological function following spinal cord injury, underscoring the vital nature of Fisetin as a potential therapeutic agent.
神经炎症是脊髓损伤后继发性损伤级联反应的一个关键组成部分,涉及促炎细胞的激活和炎症介质的释放。神经炎症的解决与细胞自噬密切相关。本研究旨在探讨天然抗炎化合物漆黄素通过调节促炎细胞中的自噬,改善神经炎症并提供脊髓损伤保护的潜力。
利用大鼠 T10 脊髓损伤模型,设置不同的治疗组(假手术组、漆黄素治疗组和漆黄素联合自噬抑制剂组),同时利用脂多糖(LPS)刺激的小胶质细胞激活和与神经元共培养的体外模型,我们采用转录组测序、组织学评估(免疫荧光染色等)、分子分析(PCR、WB、ELISA 等)和行为评估等技术,以区分神经炎症、自噬、神经元凋亡和神经功能恢复方面的差异。
漆黄素显著增强了损伤脊髓组织中的自噬活性,对脊髓损伤大鼠的神经功能恢复至关重要。漆黄素的自噬依赖性作用与损伤部位神经元凋亡的减少有关。该治疗通过自噬依赖性途径减少了 CD68+和 iNOS+细胞的数量,并降低了促炎细胞因子 IL-6 和 TNF-α的水平。漆黄素预处理可减轻 LPS 诱导的小胶质细胞的促炎极化,而自噬抑制部分阻断了这种保护作用。漆黄素在损伤脊髓和促炎小胶质细胞中诱导的自噬与 AMPK 的显著激活和 mTOR 的抑制有关。
漆黄素通过 AMPK-mTOR 信号通路协调促炎小胶质细胞中的增强自噬,从而减轻神经炎症并减少神经炎症对神经元的凋亡作用。这种机制上的见解对脊髓损伤后神经功能的保护和恢复具有重要意义,突显了漆黄素作为一种潜在治疗剂的重要性。