Agrin/Rapsyn对脊髓损伤大鼠小胶质细胞抑炎作用的实验研究  

Research on Anti-inflammatory Effect of Agrin/Rapsyn on Microglia in Rats with Spinal Cord Injury

作　　者：王晓陆[1,2] 尹宗生[1] Wang Xiaolu;Yin Zongsheng(Department of Orthopaedic,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;Department of Spinal Surgery,Hefei First People's Hospital,Hefei 230031,China)

机构地区：[1]安徽医科大学第一附属医院骨科,合肥230022 [2]合肥市第一人民医院脊柱外科,合肥230031

出　　处：《华中科技大学学报(医学版)》2025年第1期60-66,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.82372411)。

摘　　要：目的研究聚集蛋白(Agrin)、支架蛋白(Rapsyn)激活α4β2尼古丁乙酰胆碱受体(α4β2 nAChR)抑制SD大鼠的损伤脊髓组织中小胶质细胞与巨噬细胞的炎性作用。方法以4周龄SD大鼠制作脊髓损伤模型,巨噬细胞培养后以Agrin与Rapsyn孵育,流式细胞术检测小胶质细胞与巨噬细胞标记物的变化及免疫共沉淀(Co-IP)检测两种蛋白的互作,慢病毒注射脊髓损伤(SCI)模型后qRT-PCR、免疫组织化学(IHC)检测脊髓组织中炎性细胞标记物的变化。结果脊髓损伤后1~14 d小胶质细胞与巨噬细胞呈增高趋势(均P<0.01),3~7 d增长最快;经qRT-PCR分析得出SCI后炎性小胶质细胞因子(CD68)和可溶性炎性因子(IL-1β和CCL3)上调。经过鞘内注射Agrin和Rapsyn过表达的慢病毒,在0~1周的观察中,A组和R组有轻度减弱这些标记物的上调,而A/R组显著减弱了这些标记的上调(均P<0.01);经IHC分析可见,在A/R组比A组和R组小胶质细胞标记细胞数量明显减少(均P<0.01)。结论SCI后Agrin、Rapsyn激活α4β2 nAChR而产生对小胶质细胞及炎性巨噬细胞的抑制作用,从而可能阻断或减缓脊髓损伤的进展。Objective To investigate the inhibitory effects of aggregation protein(Agrin)and scaffold protein(Rapsyn)on the inflammatory responses of microglia and macrophages in injured spinal cord tissue of SD rats through activation ofα4β2 nicotinic acetylcholine receptor(α4β2 nAChR).Methods A spinal cord injury(SCI)model was established by using 4-week-old SD rats.Microglia was cultured,followed by incubation with Agrin and Rapsyn.Flow cytometry was utilized to detect changes in markers such as microglia and macrophages,while Co-IP was employed to detect the interaction between the two proteins.The qRT-PCR and IHC were used to identify changes in inflammatory cell marker expression in the spinal cord tissue of SCI models after lentivirus injection.Results The number of microglia and macrophages increased significantly from Day 1 to Day 14 after SCI(all P<0.01),with the most substantial increase observed from Day 3 to Day 7.The qRT-PCR analysis revealed that inflammatory microglial cytokine(CD68)and soluble inflammatory cytokines(IL-1βand CCL3)were upregulated after SCI.After injection of lentivirus expressing Agrin and Rapsyn,both A and R groups exhibited a slight reduction in the upregulation of these markers during the observation period of 0-1 week,whereas the A/R group demonstrated a significant reduction in their upregulation(all P<0.01).IHC analysis indicated that the number of labeled microglial cells in the A/R group was notably lower than that in either A group or R group(all P<0.01).Conclusion After SCI,Agrin and Rapsyn activateα4β2 nAChR and has inhibitory effects on microglia and inflammatory macrophages,which may block or slow down the progression of spinal cord injury.

关 键 词：脊髓损伤 AGRIN RAPSYN 小胶质细胞 抑炎作用 

分 类 号：Q74[生物学—分子生物学]