上海交通大学学报(医学版) ›› 2021, Vol. 41 ›› Issue (1): 35-41.doi: 10.3969/j.issn.1674-8115.2021.01.006

• 基础研究 • 上一篇    下一篇

法舒地尔联合过表达趋化因子受体5的间充质干细胞对实验性自身免疫性脑脊髓炎的治疗作用

李艳花1(), 闫亚平2, 刘晓琴1, 席国萍1, 宋国斌1, 肖保国3, 马存根1,4()   

  1. 1.山西大同大学脑科学研究所,神经炎症及变性疾病基础与应用研究山西省重点实验室,大同 037009
    2.陕西师范大学生命科学学院,西安 710062
    3.复旦大学附属华山医院神经病学研究所,上海 200040
    4.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,晋中 030619
  • 出版日期:2021-01-28 发布日期:2021-02-22
  • 通讯作者: 马存根 E-mail:278912829@qq.com;macungen2001@163.com
  • 作者简介:李艳花(1977—),女,副教授,博士;电子信箱:278912829@qq.com
  • 基金资助:
    山西省回国留学人员科研资助项目(HGKY2019089);大同市国际合作重点研发计划项目(2019123);中枢神经炎症变性疾病新药创制省市共建山西省重点实验室培育基地(201805D111009)

Therapeutic effects of the combination of fasudil and C-C chemokine receptor type 5-transducted mesenchymal stem cells on experimental autoimmune encephalomyelitis

Yan-hua LI1(), Ya-ping YAN2, Xiao-qin LIU1, Guo-ping XI1, Guo-bin SONG1, Bao-guo XIAO3, Cun-gen MA1,4()   

  1. 1.Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases, Institute of Brain Science, Shanxi Datong University, Datong 037009, China
    2.College of Life Science, Shaanxi Normal University, Xi'an 710062, China
    3.Institute of Neurology, Huashan Hospital, Fudan University, Shanghai 200040, China
    4.The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine, Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, China
  • Online:2021-01-28 Published:2021-02-22
  • Contact: Cun-gen MA E-mail:278912829@qq.com;macungen2001@163.com
  • Supported by:
    Funding Information] Research Project Supported by Shanxi Scholarship Council of China(HGKY2019089);International Key Research & Development Cooperation Plan of Datong(2019123);Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases(201805D111009)

摘要:

目的·研究法舒地尔(fasudil)联合过表达趋化因子受体5(C-C chemokine receptor type 5,CCR5)的间充质干细胞(mesenchymal stem cells,MSCs)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的治疗效果。方法·小鼠经髓鞘少突胶质细胞糖蛋白第35~55位肽片段(myelin oligodendrocyte glycoprotein peptide fragment 35~55,MOG35-55)免疫后,随机分为EAE模型组(腹腔注射生理盐水)、fasudil干预组(腹腔注射fasudil)、CCR5-MSCs干预组(鼻腔给予CCR5-MSCs)、fasudil联合CCR5-MSCs干预组(鼻腔给予CCR5-MSCs,腹腔注射fasudil),每组8只。MOG35-55免疫后,每日记录EAE临床症状评分。于免疫后第28日处死小鼠,固蓝染色观察髓鞘完整性,免疫荧光染色分析脊髓组织中神经干细胞和少突胶质前体细胞的产生,Western blotting检测脊髓组织中神经营养因子的表达。结果·与其他3组相比,fasudil联合CCR5-MSCs干预组EAE临床症状评分降低(P=0.000,P=0.007,P=0.005),脱髓鞘减轻(P=0.000,P=0.009,P=0.008),脊髓组织中神经干细胞和少突胶质前体细胞的产生增多(均P=0.000),脑源性神经营养因子、胶质细胞源性神经营养因子、神经生长因子和神经营养因子3的表达增加(均P=0.000)。结论·Fasudil联合CCR5-MSCs对EAE的治疗效果较fasudil或CCR5-MSCs单独治疗更为显著,可能与其促进髓鞘再生和增加神经营养因子产生相关。

关键词: 法舒地尔, 趋化因子受体5, 间充质干细胞, 实验性自身免疫性脑脊髓炎

Abstract:

Objective·To study the therapeutic effect of the combination of fasudil and C-C chemokine receptor type 5-transducted mesenchymal stem cells (CCR5-MSCs) on experimental autoimmune encephalomyelitis (EAE).

Methods·The mice immunized with myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG35-55) were randomly divided into EAE model group (intraperitoneal injection of normal saline), fasudil intervention group (intraperitoneal injection of fasudil), CCR5-MSCs intervention group (nasal administration of CCR5-MSCs), and fasudil combined with CCR5-MSCs intervention group (intranasal administration of CCR5-MSCs, intraperitoneal injection of fasudil), with 8 mice in each group. The clinical score of EAE mice were checked every day after MOG35-55 immunization. All mice were sacrificed on the 28th day after the first immunization. The myelin integrity was observed by luxol fast blue staining. The production of neural stem cells and oligodendrocyte precursor cells in spinal cord tissue was analyzed by immunofluorescence staining. The expressions of neurotrophic factors in spinal cord tissue were detected by Western blotting.

Results·Compared with the other three groups, the clinical score of EAE in the fasudil combined with CCR5-MSCs intervention group was lower (P=0.000, P=0.007, P=0.005), and the demyelination was reduced (P=0.000, P=0.009, P=0.008). The production of neural stem cells and oligodendrocyte precursor cells in spinal cord tissue was increased (all P=0.000), and the expression of brain-derived neurotrophic factor, glial cell-derived neurotrophic factor, nerve growth factor, and neurotrophin-3 increased (all P=0.000).

Conclusion·Fasudil combined with CCR5-MSCs is more effective than fasudil or CCR5-MSCs in the treatment of EAE, which may be related to the induction of remyelination and the production of the neurotrophic factor.

Key words: fasudil, C-C chemokine receptor type 5 (CCR5), mesenchymal stem cell (MSC), experimental autoimmune encephalomyelitis (EAE)

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