中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (7): 1711-1719.doi: 10.12307/2026.047

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间充质干细胞来源外泌体减轻脓毒症脑病小鼠血脑屏障损伤

夏林枫1，2，王  露3，龙乾发4，唐荣武2，罗浩东5，汤  轶1，钟  俊2，刘  阳1，2   

  1. 1川北医学院临床医学院，四川省南充市   637000；绵阳市第三人民医院(四川省精神卫生中心)，2神经外科，3放射科，四川省绵阳市   621000；4西安市中心医院神经外科，陕西省西安市   710003；5西南医科大学临床医学院，四川省泸州市   646000
  • 收稿日期:2024-12-19 修回日期:2025-04-22 接受日期:2025-05-29 出版日期:2026-03-08 发布日期:2025-08-19
  • 通讯作者: 刘阳，博士，主任医师，川北医学院临床医学院，四川省南充市 637000；绵阳市第三人民医院(四川省精神卫生中心)神经外科， 四川省绵阳市 621000
  • 作者简介:夏林枫，男，1997年生，重庆市人，汉族，川北医学院在读硕士，医师，主要从事脑血管病、间充质干细胞研究。
  • 基金资助:
    绵阳市卫生健康委员会支持项目(202201)，项目负责人：刘阳；四川省医学会支持项目(Q22020)，项目负责人：钟俊

Human umbilical cord mesenchymal stem cell-derived exosomes alleviate blood-brain barrier damage in mice with septic encephalopathy

Xia Linfeng1, 2, Wang Lu3, Long Qianfa4, Tang Rongwu2, Luo Haodong5, Tang Yi1, Zhong Jun2, Liu Yang1, 2   

  1. 1School of Clinical Medicine, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; 2Department of Neurosurgery, 3Department of Radiology, The Third People’s Hospital of Mianyang (Sichuan Mental Health Center), Mianyang 621000, Sichuan Province, China; 4Department of Neurosurgery, Xi’an Central Hospital, Xi’an 710003, Shaanxi Province, China; 5School of Clinical Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2024-12-19 Revised:2025-04-22 Accepted:2025-05-29 Online:2026-03-08 Published:2025-08-19
  • Contact: Liu Yang, MD, Chief physician, School of Clinical Medicine, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Department of Neurosurgery, The Third People’s Hospital of Mianyang (Sichuan Mental Health Center), Mianyang 621000, Sichuan Province, China
  • About author:Xia Linfeng, Master candidate, Physician, School of Clinical Medicine, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; Department of Neurosurgery, The Third People’s Hospital of Mianyang (Sichuan Mental Health Center), Mianyang 621000, Sichuan Province, China
  • Supported by:
    Mianyang Municipal Health Commission Support Project, No. 202201 (to LY); Sichuan Medical Association Support Project, No. Q22020 (to ZJ) 

PDF

0

摘要：

文题释义：

外泌体：富含miRNA及蛋白质的小囊泡，可以通过旁分泌参与细胞间通讯及免疫调节。由间充质干细胞分泌的外泌体在抗炎、抑凋亡和促进血管新生等多个方面发挥出重要作用。
血脑屏障：由脑内毛细血管内皮细胞间的紧密连接构成，可阻止有害物质进入中枢神经系统，保护脑神经元。中枢神经系统疾病常引起血脑屏障破坏和功能紊乱，加剧脑水肿及中枢神经系统炎症，造成患者预后不良。

摘要
背景：研究证实间充质干细胞来源外泌体作为细胞间通信递质可抑制神经炎症和促进血管新生，目前关于间充质干细胞来源外泌体减轻脓毒症脑病血脑屏障损伤的作用机制研究较少。
目的：探究人脐带间充质干细胞来源外泌体对脓毒症脑病小鼠血脑屏障的保护作用及作用机制。 
方法：采用超速离心法从人脐带间充质干细胞培养基中分离外泌体。51只C57/BL6小鼠随机分为假手术组(n=17)、模型组(n=17)、治疗组(n=17)，后2组腹腔注射脂多糖构建脓毒症脑病模型，治疗组在造模后0.5 h尾静脉注射50 μg外泌体。24 h后，采用小鼠干湿比重法和伊文思蓝法检测小鼠脑组织含水量和血脑屏障通透性，尼氏染色检测小鼠海马区神经元损伤情况，免疫荧光染色观察脑皮质组织中闭锁小带蛋白1和咬合蛋白荧光强度，Western blot检测脑皮质组织中肿瘤坏死因子α、白细胞介素1β、闭锁小带蛋白1、咬合蛋白、高迁移率族蛋白B1、Toll样受体4、核因子κB及基质金属蛋白酶9表达水平。

结果与结论：①与假手术组相比，模型组小鼠脑组织含水量和伊文思蓝含量显著升高(P < 0.05)；与模型组相比，治疗组小鼠脑组织含水量和伊文思蓝含量显著降低(P < 0.05)；②尼式染色结果显示，与假手术组相比，模型组小鼠海马组织CA1区神经元细胞形态不规则、排列不整齐，神经元数量显著降低(P < 0.05)；与模型组相比，治疗组小鼠海马组织CA1区神经元细胞形态规则、排列整齐，神经元数量显著增加(P < 0.05)；③免疫荧光染色结果显示，与假手术组相比，模型组小鼠脑皮质组织中闭锁小带蛋白1和咬合蛋白荧光强度显著降低；与模型组相比，治疗组小鼠脑皮质组织中闭锁小带蛋白1和咬合蛋白荧光强度显著升高；④Western blot结果显示，与假手术组相比，模型组小鼠脑皮质组织中肿瘤坏死因子α、白细胞介素1β、高迁移率族蛋白B1、Toll样受体4、核因子κB及基质金属蛋白酶9表达水平显著增加(P < 0.05)；与模型组相比，治疗组小鼠脑皮质组织中肿瘤坏死因子α、白细胞介素1β、高迁移率族蛋白B1、Toll样受体4、核因子κB及基质金属蛋白酶9表达水平显著降低(P < 0.05)；⑤结果表明，人脐带间充质干细胞来源外泌体可以减轻脓毒症脑病小鼠血脑屏障损伤，降低血脑屏障通透性，其作用机制与外泌体抑制高迁移率族蛋白B1/Toll样受体4/基质金属蛋白酶9相关炎症通路有关。

https://orcid.org/0009-0006-2855-2837 (夏林枫) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 人脐带间充质干细胞, 外泌体, 脂多糖, 脓毒症脑病, 血脑屏障, 高迁移率族蛋白B1, 基质金属蛋白酶9, 神经炎症

Abstract: BACKGROUND: Mesenchymal stem cell-derived exosomes, as intercellular communication mediators, have been shown to inhibit neuroinflammation and promote angiogenesis. However, there are few studies on the mechanism of action of mesenchymal stem cell-derived exosomes in alleviating the damage of the blood-brain barrier in septic encephalopathy.
OBJECTIVE: To explore the protective effect and mechanism of exosomes derived from human umbilical cord mesenchymal stem cells on the blood-brain barrier of mice with septic encephalopathy
METHODS: Exosomes were isolated from human umbilical cord mesenchymal stem cell culture medium by ultracentrifugation. Fifty-one C57/BL6 mice were randomly divided into sham operation group (n=17), model group (n=17), and treatment group (n=17). The latter two groups were intraperitoneally injected with lipopolysaccharide to establish the sepsis encephalopathy model. The treatment group was injected with 50 μg exosomes 0.5 hours after modeling. After 24 hours, the water content of brain tissue and the permeability of blood-brain barrier in mice were detected by mouse dry wet gravity method and Evans blue method. The neuronal damage in the mouse hippocampus was detected by Nissl staining. The fluorescence intensity of zonula occludens protein 1 and occludin protein in cerebral cortex was observed by immunofluorescence staining. The expression levels of tumor necrosis factor-α, interleukin-1β, zonula occludens protein 1, occludin protein, high mobility group protein B1, toll like receptor 4, nuclear factor κB, and matrix metalloproteinase 9 in cerebral cortex were detected by western blot assay. 
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, the brain water content and Evans blue content in the model group were significantly increased (P < 0.05), while compared with the model group, the brain water content and Evans blue content in the treatment group were significantly decreased (P < 0.05). (2) Nissl staining results showed that compared with the sham operation group, the morphology and arrangement of neurons in hippocampal CA1 area of the model group were irregular, and the number of neurons was significantly decreased (P < 0.05). Compared with the model group, the morphology and arrangement of neurons in hippocampal CA1 area of the treatment group were regular, and the number of neurons was significantly increased (P < 0.05). (3) Immunofluorescence staining results showed that compared with the sham operation group, the fluorescence intensity of zonula occludens protein 1 and occludin protein in the cerebral cortex of the model group was significantly decreased, and compared with the model group, the fluorescence intensity of zonula occludens protein 1 and occludin protein in the cerebral cortex of the treatment group was significantly increased. (4) Western blot assay results showed that compared with the sham operation group, the expression levels of tumor necrosis factor-α, interleukin-1β, high mobility group protein B1, toll like receptor 4, nuclear factor κB, and matrix metalloproteinase-9 were significantly increased in the cerebral cortex of the model group (P < 0.05), while the expression levels of tumor necrosis factor-α, interleukin-1β, high mobility group protein B1, toll like receptor 4, nuclear factor kappa B, and matrix metalloproteinase-9 were significantly decreased in the cerebral cortex of the treatment group (P < 0.05). (5) The results showed that human umbilical cord mesenchymal stem cell-derived exosomes could reduce the damage of blood-brain barrier and the permeability of blood-brain barrier in mice with sepsis encephalopathy, and its mechanism was related to the inhibition of high mobility group protein B1/toll like receptor 4/matrix metalloproteinase 9 related inflammatory pathway by exosomes. 

Key words: human umbilical cord mesenchymal stem cell, exosome, lipopolysaccharide, septic encephalopathy, blood-brain barrier, high mobility group protein B1, matrix metalloproteinase 9, neuroinflammation

中图分类号: