中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (33): 8712-8720.doi: 10.12307/2026.465

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

急性缺血性脑卒中模型小鼠继发性脱髓鞘损伤和炎症反应的机制

张紫薇1，蒲  萌1，汤亿彬1，郑璐璐1，梁亚杰1，刘  健1，陈  莹1，王  青1，李彦青1，李艳花1，2   

  1. 1山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心，山西省晋中市   030619；2山西大同大学医学院，山西省大同市   037009
  • 收稿日期:2025-10-17 修回日期:2026-02-27 出版日期:2026-11-28 发布日期:2026-06-15
  • 通讯作者: 王青，副教授，硕士生导师，山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心，山西省晋中市 030619
  • 作者简介:张紫薇，女，1997年生，硕士，主要从事中西医结合防治中枢神经系统炎性变性疾病研究。
  • 基金资助:
    山西省回国留学人员科研资助项目(2022-165)，项目负责人：王青；山西省中医药管理局中医药创新团队(zyytd2024039)，项目负责人：王青；山西省中医药管理局科研课题(2023ZYYB040)，项目负责人：王青

Mechanisms of secondary demyelinating injury and inflammatory response in mice with acute ischemic stroke

Zhang Ziwei1, Pu Meng1, Tang Yibin1, Zheng Lulu1, Liang Yajie1, Liu Jian1, Chen Ying1, Wang Qing1, Li Yanqing1, Li Yanhua1, 2   

  1. 1The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis, State Administration of Traditional Chinese Medicine/Neurobiology Research Center, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; 2School of Medicine, Shanxi Datong University, Datong 037009, Shanxi Province, China
  • Received:2025-10-17 Revised:2026-02-27 Online:2026-11-28 Published:2026-06-15
  • Contact: Wang Qing, Associate professor, Master’s supervisor, The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis, State Administration of Traditional Chinese Medicine/Neurobiology Research Center, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • About author:Zhang Ziwei, MS, The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis, State Administration of Traditional Chinese Medicine/Neurobiology Research Center, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • Supported by:
    Shanxi Province Research Support Program for Returning Overseas Scholars, No. 2022-165 (to WQ); Shanxi Provincial Administration of Traditional Chinese Medicine Innovation Team, No. zyytd2024039 (to WQ); Shanxi Provincial Administration of Traditional Chinese Medicine Research Project, No. 2023ZYYB040 (to WQ) 

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摘要：


文题释义：
脱髓鞘：分为原发性脱髓鞘和继发性脱髓鞘，多发生于神经退行性疾病和血管损伤疾病；也可以根据部位分为中枢神经系统脱髓鞘和周围神经系统脱髓鞘。脱髓鞘的发生会使认知功能发生减退，感觉、运动功能受损，表现为记忆力下降、言语不清、肢体无力等，严重时导致死亡。
小胶质细胞：一种神经胶质细胞，负责大脑中枢神经系统的稳态，具有免疫防御、损伤修复、神经发育等作用，可以被激活极化为促炎表型(M1)和抗炎表型(M2)。

背景：远端大脑中动脉闭塞小鼠会发生继发性脱髓鞘损伤。在急性期，脱髓鞘损伤进程与小胶质细胞炎症反应密切相关。小胶质细胞具有吞噬作用，通过相关机制清除损伤后降解的髓鞘碎片，修复损伤区域微环境。
目的：利用电凝法建立缺血性脑卒中小鼠模型，探讨脱髓鞘、炎症反应及相关机制。
方法：首先将昆明小鼠随机分为假手术组、电凝组，电凝组小鼠再分为电凝12 h及电凝1，3，5，7 d组，每组9只。以假手术组为对照，通过Bederson评分和Y迷宫实验比较小鼠的运动和记忆能力；TTC染色观察梗死区域；固蓝染色观察大脑皮质髓鞘缺失程度；免疫荧光染色检测髓鞘碱性蛋白、降解髓鞘碱性蛋白、离子钙结合接头分子1、Janus激酶2、信号转导与转录激活因子3表达；Western blot 检测Janus激酶2、信号转导与转录激活因子3蛋白表达；ELISA检测白细胞介素6、肿瘤坏死因子α、白细胞介素1β、干扰素γ、白细胞介素10水平。通过以上实验分析电凝组小鼠的髓鞘损伤和炎性反应机制。
结果与结论：采用电凝法成功构建缺血性脑卒中小鼠模型。以假手术组为对照，①各电凝组小鼠出现不同程度的运动障碍，表现为同一时间内总路程出现显著性差异；②在电凝1 d时，TTC染色缺血区域(P < 0.001)、固蓝染色大脑皮质脱髓鞘区域(P < 0.001)达到最大面积；③免疫荧光染色显示髓鞘碱性蛋白在电凝1 d时显著减少(P < 0.05)，降解髓鞘碱性蛋白在电凝1 d时明显增多(P < 0.001)，离子钙结合接头分子1在电凝3-5 d时显著增多(P < 0.001)，Janus激酶2在电凝1 d时显著增多(P < 0.01)，信号转导与转录激活因子3在电凝1-3 d时显著增多(P < 0.01)；④ELISA结果显示炎症因子水平在电凝后显著升高；⑤Western blot结果显示，电凝组的Janus激酶2、信号转导与转录激活因子3蛋白表达显著升高。结果表明：电凝法导致模型小鼠中枢神经系统脱髓鞘，影响运动和记忆功能，引发大脑炎症反应，激活Janus激酶2-信号转导与转录激活因子3通路，该通路可能参与小胶质细胞吞噬髓鞘碎片。
https://orcid.org/0000-0003-1284-4705 (王青) 


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: 脱髓鞘, 缺血性脑卒中, 小胶质细胞, 神经炎症, Janus激酶2-信号转导与转录激活因子3, 吞噬作用

Abstract: BACKGROUND: Distal middle cerebral artery occlusion in mice leads to secondary demyelinating lesions. In the acute phase, the progression of demyelinating injury is closely related to the inflammatory response of microglia. Microglia possess phagocytic functions and clear degraded myelin debris through relevant mechanisms post-injury, thereby repairing the microenvironment of the damaged area.
OBJECTIVE: To establish a mouse model of ischemic stroke using electrocoagulation and investigate demyelination, inflammatory responses, and related mechanisms.
METHODS: Kunming mice were randomly divided into a sham operation group and an electrocoagulation group. The electrocoagulation group was further subdivided into 12-hour, 1-day, 3-day, 5-day, and 7-day subgroups, with nine mice in each group. Using the sham group as the control, motor and memory abilities were compared via Bederson scoring and Y-maze tests. 2,3,5-Triphenyl tetrazolium chloride staining was used to observe the infarct area, while Luxol fast blue staining was employed to detect the degree of myelin loss in the cerebral cortex. Immunofluorescence staining was performed to detect the expression of myelin basic protein, degraded myelin basic protein, ionized calcium-binding adapter molecule 1, Janus kinase 2 (JAK2), and signal transducer and activator of transcription 3 (STAT3). Western blot was used to measure JAK2 and STAT3 protein expression, and ELISA was conducted to quantify levels of interleukin-6, tumor necrosis factor-α, interleukin-1β, interferon-γ, and interleukin-10. These experiments were employed to analyze the mechanisms of myelin injury and inflammatory responses in the model mice.
RESULTS AND CONCLUSION: The ischemic stroke model was successfully established in mice using electrocoagulation. Compared with the sham group, the following results were observed: (1) Mice in the electrocoagulation subgroups exhibited varying degrees of motor dysfunction, with significant differences in total distance traveled within the same time frame. (2) At 1 day of electrocoagulation, the ischemic area identified by 2,3,5-triphenyl tetrazolium chloride staining (P < 0.001) and the demyelinated area in the cerebral cortex detected by Luxol fast blue staining (P < 0.001) reached their maximum extent. (3) Immunofluorescence staining showed that myelin basic protein significantly decreased at 1 day of electrocoagulation (P < 0.05), degraded myelin basic protein significantly increased at 1 day of electrocoagulation (P < 0.001), ionized calcium-binding adapter molecule 1 significantly increased at 3-5 days of electrocoagulation (P < 0.001), JAK2 significantly increased at 1 day of electrocoagulation (P < 0.01), and STAT3 significantly increased at 1-3 days of electrocoagulation (P < 0.01). (4) ELISA results indicated that inflammatory factor levels significantly increased after electrocoagulation. (5) Western blot results revealed significantly elevated protein expression of JAK2 and STAT3 in the electrocoagulation group. These findings indicate that electrocoagulation induces central nervous system demyelination, impairs motor and memory functions, triggers cerebral inflammatory responses, and activates the JAK2-STAT3 pathway, which may be involved in microglial phagocytosis of myelin debris.


Key words: demyelination, ischemic stroke, microglia, neuroinflammation, Janus kinase 2-signal transducer and activator of transcription 3, phagocytosis

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