脂肪间充质干细胞来源外泌体对脊髓损伤大鼠巨噬细胞极化及胶质瘢痕形成的影响  

Effects of Exosomes Derived from Adipose-Derived Mesenchymal Stem Cells on Macrophage Polarization and Glial Scar Formation in Rats with Spinal Cord Injury

作　　者：陈浩贤 严利军 安永刚 杨明坤[2] Chen Haoxian;Yan Lijun;An Yonggang(Orthopedics Department of Sichuan Modern Hospital,Chengdu,Sichuanu 610045,China)

机构地区：[1]四川现代医院骨科,四川成都610045 [2]巴中市中心医院骨科,四川巴中636600

出　　处：《四川医学》2022年第4期333-338,共6页Sichuan Medical Journal

摘　　要：目的探讨脂肪间充质干细胞(ADMSC)来源外泌体对脊髓损伤大鼠巨噬细胞极化及胶质瘢痕形成的影响。方法提取ADMSC来源外泌体并鉴定。选取36只大鼠,分为假手术组(仅切除T9椎板+注射等量生理盐水)、脊髓损伤组(建模+注射等量生理盐水)、外泌体组(建模+注射100μg/kg外泌体),每组各12只,采用钳夹脊髓法建立脊髓损伤大鼠模型。使用BBB评分评估大鼠后肢运动功能,酶联免疫(ELISA)法检测血清白细胞介素(IL)-1β、IL-4、IL-6、IL-10水平,对脊髓组织行尼氏染色,Western Blot法检测脊髓组织中诱导型一氧化氮合酶(iNOS)、精氨酸酶-1(Arg-1)、胶质纤维酸性蛋白(GFAP)、波形蛋白(vimentin)蛋白表达。结果经透射电镜观察及CD63、Alix蛋白检测鉴定,成功提取到ADMSC来源外泌体。与假手术组相比,脊髓损伤组大鼠BBB评分显著降低(P<0.05),血清IL-1β、IL-6、IL-4、IL-10水平及脊髓组织中iNOS、Arg-1、GFAP、vimentin蛋白水平均显著升高(P<0.05),脊髓组织中神经细胞严重损伤,可见大量胶质瘢痕组织堆积;与脊髓损伤组相比,外泌体组大鼠BBB评分、血清IL-4、IL-10水平及脊髓组织中Arg-1蛋白水平显著升高(P<0.05),血清IL-1β、IL-6水平及脊髓组织中iNOS、GFAP、vimentin蛋白水平显著降低(P<0.05),脊髓组织中神经细胞损伤及胶质瘢痕组织堆积减轻。结论ADMSC来源外泌体可改变脊髓损伤大鼠巨噬细胞极化方向,并抑制胶质瘢痕形成。Objective To investigate the effects of adipose-derived mesenchymal stem cells(ADMSC)-derived exosomes on the polarization of macrophages and the formation of glial scars in rats with spinal cord injury.Methods The ADMSC-derived exosomes were extracted and identified.The 36 rats were divided into sham operation group(removal of T9 lamina+injection of equal amount of saline),spinal cord injury group(modeling+injection of equal amount of saline),and exosomes group(modeling+injection of 100μg/kg exosomes),12 rats in each group,the spinal cord clamp method was used to establish a rat model of spinal cord injury.The BBB score was used to evaluate the motor function of the hind limbs of rats,enzyme-linked immunoassay(ELISA)method was used to detect the levels of serum interleukin(IL)-1β,IL-4,IL-6,and IL-10,Nissl staining was conducted on the spinal cord tissue,western blot was used to detect the expression of inducible nitric oxide synthase(iNOS),arginase-1(Arg-1),glial fibrillary acidic protein(GFAP)and vimentin(vimentin)proteins in spinal cord tissue.Results After transmission electron microscopy observation,CD63 and Alix protein detection and identification,ADMSC-derived exosomes were successfully extracted.Compared with the sham operation group,the BBB score of rats in the spinal cord injury group was significantly lower(P<0.05),the serum IL-1β,IL-6,IL-4,IL-10 levels and iNOS,Arg-1,GFAP,and vimentin protein levels in spinal cord tissue were significantly increased(P<0.05),the nerve cells in the spinal cord tissue were seriously damaged,and a large amount of glial scar tissue could be seen accumulation;compared with the spinal cord injury group,the BBB score,serum IL-4 and IL-10 levels and the Arg-1 protein level in the spinal cord tissue of the exosomes group were significantly increased(P<0.05),the serum IL-1βand IL-6 levels and iNOS,GFAP,and vimentin protein levels in spinal cord tissue were significantly reduced(P<0.05),the nerve cell damage and glial scar tissue accumulation in spinal cord tissues were redu

关 键 词：脂肪间充质干细胞 外泌体 脊髓损伤 大鼠 巨噬细胞极化 胶质瘢痕形成 

分 类 号：R-332[医药卫生] R744