Sun Yan, Gao Sheng-Qing, Wang Xue, Li Tao, Han Yan-Ling, Miao Shu-Hao, Zhao Ran, Zheng Xiao-Bo, Qiu Jia-Yin, Jin Wang-Xuan, Gao Chao-Chao, Zhou Meng-Liang
Department of Neurosurgery, Jinling Hospital, Jinling School of Clinical Medicine, Nanjing Medical University, Nanjing, China.
Department of Neurosurgery, Nanjing Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Brain Res. 2025 May 15;1855:149560. doi: 10.1016/j.brainres.2025.149560. Epub 2025 Mar 10.
Externally caused traumatic brain injury (TBI) poses a woeful worldwide health concern, bringing about disability, death, and prolonged neurological impairment. Increased galectin-3 levels have been linked to unfavorable outcomes in several neurological conditions. This study explores the role of galectin-3 in TBI, specifically examining its contribution to neuroinflammation.
BV2 microglia cells treated with lipopolysaccharide (LPS) and a mouse model of TBI were applied to investigate the impact of galectin-3 on neuroinflammation following TBI. Western blotting and immunofluorescence labeling were applied for evaluating protein levels and colocalization. Adeno-associated virus (AAV) that targets microglia was used to knock down galectin-3 in microglia. Nissl staining and the modified neurologic severity score were employed in evaluating neural survival and neurological function, and the cognitive impairment following TBI was assessed by the Y-Maze and Morri water maze test.
Galectin-3 expression was shown to rise dramatically after TBI, peaking between days five and seven. In vitro, BV2 cells treated with LPS showed reduced NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome activation when galectin-3 was inhibited. In LPS-activated microglia, galectin-3 inhibition specifically decreased the expression of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), p-NF-κB, NLRP3, Apoptosis-associated speck-like protein containing a CARD (ASC), caspase-1, and Gasdermin D (GSDMD). Injection with AAV containing siRNA to knock down galectin-3 in microglia was operated on mice in vivo. Following TBI, this knockdown led to reduced NLRP3 inflammasome activation, neuronal death, neurological impairments and cognitive impairment.
Our foundings indicate that modulating microglia-derived galectin-3 following TBI to reduce neuroinflammation could serve as a promising therapeutic strategy.
外部导致的创伤性脑损伤(TBI)是一个令人悲痛的全球健康问题,会导致残疾、死亡和长期神经功能障碍。半乳糖凝集素-3水平升高与多种神经系统疾病的不良预后有关。本研究探讨半乳糖凝集素-3在TBI中的作用,特别研究其对神经炎症的影响。
应用脂多糖(LPS)处理的BV2小胶质细胞和TBI小鼠模型,研究半乳糖凝集素-3对TBI后神经炎症的影响。采用蛋白质印迹法和免疫荧光标记法评估蛋白水平和共定位情况。使用靶向小胶质细胞的腺相关病毒(AAV)敲低小胶质细胞中的半乳糖凝集素-3。采用尼氏染色和改良神经功能缺损评分评估神经存活和神经功能,通过Y迷宫和莫里斯水迷宫试验评估TBI后的认知障碍。
TBI后半乳糖凝集素-3表达显著升高,在第5天至第7天达到峰值。在体外,当半乳糖凝集素-3受到抑制时,用LPS处理的BV2细胞显示核苷酸结合寡聚化结构域样受体热蛋白结构域相关蛋白3(NLRP3)炎性小体激活减少。在LPS激活的小胶质细胞中,半乳糖凝集素-3抑制特异性降低Toll样受体4(TLR4)、核因子-κB(NF-κB)、p-NF-κB、NLRP3、含半胱天冬酶激活和招募结构域的凋亡相关斑点样蛋白(ASC)、半胱天冬酶-1和Gasdermin D(GSDMD)的表达。在体内对小鼠注射含小干扰RNA的AAV以敲低小胶质细胞中的半乳糖凝集素-3。TBI后,这种敲低导致NLRP3炎性小体激活减少、神经元死亡、神经功能障碍和认知障碍减轻。
我们的研究结果表明,TBI后调节小胶质细胞来源的半乳糖凝集素-3以减轻神经炎症可能是一种有前景的治疗策略。