神经内镜下人脐带间充质干细胞外泌体鞘内移植修复脊髓损伤的作用机制  

Action mechanism of intrathecal transplantation of human umbilical cord mesenchymal stem cell-derived exosomes for repair of spinal cord injury under neuroendoscopy

作　　者：郑伊桐 汪永新[1] 刘文 阿木吉特 秦虎[1] Zheng Yitong;Wang Yongxin;Liu Wen;Amujite;Qin Hu(First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang Uyghur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院,新疆维吾尔自治区乌鲁木齐市830054

出　　处：《中国组织工程研究》2025年第36期7743-7751,共9页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金(2021D01C339),项目负责人:秦虎。

摘　　要：背景:研究发现,人脐带间充质干细胞来源外泌体可以有效促进脊髓损伤的神经修复。目的:探讨人脐带间充质干细胞来源外泌体是否可以通过促进小胶质细胞向M2型极化减轻神经炎症促进脊髓损伤大鼠运动功能恢复。方法:将48只SD大鼠随机分为假手术组、模型组和外泌体组(n=16),采用改良Allen法建立大鼠脊髓损伤模型。外泌体组在损伤后24 h通过神经内镜鞘内注射20μL人脐带间充质干细胞来源外泌体。在造模后3,7,14,21 d,采用BBB评分法结合Rivlin斜板实验评估大鼠后肢运动功能的恢复情况,采用苏木精-伊红染色和尼氏染色检测脊髓组织损伤情况,Western blot检测脊髓组织中脑源性神经营养因子和血管内皮生长因子A蛋白表达水平,免疫荧光法检测脊髓组织中M1型标志物(诱导型一氧化氮合酶)与M2型标志物(精氨酸酶1)的表达比例,qRT-PCR和Western blot检测脊髓组织中诱导型一氧化氮合酶与精氨酸酶1的表达水平,ELISA法检测脊髓组织中促炎因子(肿瘤坏死因子α、白细胞介素1β和白细胞介素6)和抑炎因子(白细胞介素10)水平。结果与结论:(1)术后3,7,14 d,外泌体组BBB评分均高于模型组(P<0.05),术后7,14 d时外泌体组Rivlin斜板实验角度均显著高于模型组(P<0.05),苏木精-伊红染色和尼氏染色结果显示外泌体组相比于模型组脊髓组织的神经损伤减轻,术后7 d时外泌体组脑源性神经营养因子和血管内皮生长因子A表达较模型组增加(P<0.05);(2)免疫荧光实验结果显示,与模型组相比,外泌体组术后第7天病变区域的诱导型一氧化氮合酶阳性小胶质细胞明显减少,而精氨酸酶1阳性小胶质细胞明显增多(P<0.05);qRT-PCR和Western blot也证实了免疫荧光实验结果;(3)外泌体组脊髓组织中促炎因子肿瘤坏死因子α、白细胞介素1β和白细胞介素6分泌量较模型组减少(P<0.05),而抑炎因子白细胞介素10分泌量较模型组增�BACKGROUND:Human umbilical cord mesenchymal stem cell-derived exosomes were found to be effective in promoting neural repair in spinal cord injury.OBJECTIVE:To investigate whether exosomes derived from human umbilical cord mesenchymal stem cells are able to attenuate neuroinflammation and promote recovery of motor function by promoting polarization of microglia toward the M2 type.METHODS:Totally 48 SD rats were randomly divided into a sham operation group,a model group,and an exosome group(n=16 per group).A rat spinal cord injury model was established using the modified Allen method.The exosome group was injected with 20μL of human umbilical cord mesenchymal stem cell-derived exosomes intrathecally via neuroendoscopy 24 hours after injury.At 3,7,14,and 21 days after modeling,the recovery of the motor function of the hind limbs of the rats was assessed by BBB scoring method combined with Rivlin’s slant plate test.The damage of spinal cord tissues was detected by using hematoxylin-eosin staining and Nissl staining.The expression levels of brain-derived neurotrophic factor and vascular endothelial growth factor A proteins were detected by western blot assay.The expression proportion of M1-type markers(inducible nitric oxide synthase)and M2 markers(arginase-1)in the spinal cord tissues was detected by immunofluorescence method.qRT-PCR and western blot assay were used to detect the expression levels of inducible nitric oxide synthase and arginase-1 in spinal cord tissues.ELISA was utilized to detect the levels of pro-inflammatory factors(tumor necrosis factorα,interleukin 1β,and interleukin 6)and anti-inflammatory factors(interleukin 10)levels in spinal cord tissues.RESULTS AND CONCLUSION:(1)At 3,7,and 14 days postoperatively,the BBB scores of the exosome group were better than those of the model group(P<0.05).The angles of the Rivlin slanting plate experiments of the exosome group were significantly higher than those of the model group at 7 and 14 days postoperatively(P<0.05).The results of hematoxylin-eosin sta

关 键 词：脊髓损伤 人脐带间充质干细胞 小胶质细胞 外泌体 炎症 神经内镜 工程化干细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R616