芬戈莫德对神经干细胞和星形胶质细胞增殖及迁移的影响及其对小鼠脊髓损伤的保护机制

doi:10.13481/j.1671-587x.20180619

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (06): 1218-1222.doi: 10.13481/j.1671-587x.20180619

芬戈莫德对神经干细胞和星形胶质细胞增殖及迁移的影响及其对小鼠脊髓损伤的保护机制

刘林娜¹, 陈婕妤², 吕方怡², 花扣珍², 蔡大敏², 银国利², 王俊娟²

1. 温州医科大学附属第二医院检验医学科, 浙江温州 325000;
2. 杭州医学院基础医学与法医学院人体解剖与组织胚胎学教研室, 浙江杭州 310014

收稿日期:2018-05-16 出版日期:2018-11-28 发布日期:2018-11-28
通讯作者: 王俊娟,助教(Tel:0571-87692675,E-mail:wang.junjuan@foxmail.com) E-mail:wang.junjuan@foxmail.com
作者简介:刘林娜(1989-),女,浙江省温州市人,技师,在读医学硕士,主要从事小分子药物和再生医学等方面的研究。
基金资助:
浙江省科技厅自然科学基金资助课题（LY18H090011）；浙江省教育厅一般科研项目资助课题（Y201738521，Y201738494）

Effect of fingolimod on proliferation and migration of neural stem cells and astrocytes and its protective mechanism against spinal cord injury in mice

LIU Linna¹, CHEN Jieyu², LYU Fangyi², HUA Kouzhen², CAI Damin², YIN Guoli², WANG Junjuan²

1. Department of Laboratory Medicine, Second Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China;
2. Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences and Forensic Sciences, Hangzhou Medical College, Hangzhou 310014, China

Received:2018-05-16 Online:2018-11-28 Published:2018-11-28

摘要/Abstract

摘要： 目的：探讨芬戈莫德（FTY720）对神经干细胞（NSCs）和星形胶质细胞迁移及增殖的影响，阐明FTY720对小鼠脊髓损伤（SCI）的保护作用及其机制。方法：体外培养NSCs和星形胶质细胞，每种细胞随机分为对照组（采用PBS进行处理）和实验组（采用0.1μmol·L^-1FTY720进行处理）。SCI小鼠随机分为对照组（口服PBS）和实验组（口服1 mg·kg^-1·d^-1 FTY720），每组10只。采用Transwell实验检测2组NSCs和星形胶质细胞的迁移细胞数，免疫荧光染色检测NSCs的迁移细胞数和增殖神经球数，HE染色检测2组小鼠SCI空洞面积，Basso Mouse Scale （BMS）评分评估小鼠运动能力恢复情况。结果：Transwell检测，与对照组比较，实验组NSCs迁移细胞数增多（P<0.05），星形胶质细胞迁移细胞数减少（P<0.05）。免疫荧光检测，与对照组比较，实验组NSCs增殖神经球数增多（P<0.05），实验组小鼠SCI震中星形胶质细胞数减少（P<0.05）。HE染色检测，与对照组比较，SCI空洞面积缩小（P<0.05）。BMS评分，与对照组比较，实验组小鼠BMS评分升高（P<0.05）。结论：FTY720可以促进NSCs增殖和迁移，抑制星形胶质细胞迁移，减少SCI后胶质疤痕形成，促进小鼠运动功能恢复。

关键词: 脊髓损伤, 芬戈莫德, 神经干细胞, 星形胶质细胞

Abstract: Objective: To investigate the effects of fingolimod (FTY720) on the migration and proliferation of neural stem cells (NSCs) and astrocytes of the mice, and to elucidate the protective effect of FTY720 on spinal cord injury(SCI) of the mice and its mechanism.Methods: The NSCs and astrocytes were cultured in vitro and divided into control group(treated with PBS) and experiment group(treated with 0.1 μmol·L^-1 FTY720). The SCI mice were divided into control group (given PBS by oral,n=10) and expriment group (given 1 mg·kg^-1·d^-1 FTY720 by oral,n=10). The number of migration cells of NSCs and astrocytes in two groups was detected by Transwell experiment, and the number of migration cells and the number of proliferative neurospheres of NSCs were detected by immunofluorescence staining.The SCI void areas of the mice in two groups were detected by HE staining, and the dynamic recovery of motor function of the mice was assessed by Basso Mouse Scale(BMS) behavioral score.Results: The Transwell detection results showed that compared with control group, the number of migration cells of NSCs in expriment group was increased(P<0.05), and the number of migration cells of astrocytes in expriment group was decreased(P<0.05). The immunofluorescence detection results showed that compared with control group, the number of proliferative neurospheres of NSCs in expriment group was increased(P<0.05),and the number of astrocytes in damaged SCI in expriment group was decreased(P<0.05).The HE staining results showed that compared with control group, the SCI void area was decreased(P<0.05). The BMS score results showed that the BMS score of the mice in expriment group was higher than that in control group(P<0.05).Conclusion: FTY720 can promote the proliferation and migration of NSCs, inhibit the migration of astrocytes, decrease the formation of glial scar, and promote the motor function recovery of the mice after SCI.

Key words: spinal cord injury, FTY720, neural stem cells, astrocytes

中图分类号:

Q253

刘林娜, 陈婕妤, 吕方怡, 花扣珍, 蔡大敏, 银国利, 王俊娟. 芬戈莫德对神经干细胞和星形胶质细胞增殖及迁移的影响及其对小鼠脊髓损伤的保护机制[J]. 吉林大学学报(医学版), 2018, 44(06): 1218-1222.

LIU Linna, CHEN Jieyu, LYU Fangyi, HUA Kouzhen, CAI Damin, YIN Guoli, WANG Junjuan. Effect of fingolimod on proliferation and migration of neural stem cells and astrocytes and its protective mechanism against spinal cord injury in mice[J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1218-1222.

参考文献

[1] Lima R, Monteiro S, Lopes JP, et al. Systemic interleukin-4 administration after spinal cord injury modulates inflammation and promotes neuroprotection[J]. Pharmaceuticals, 2017, 10(4):83.
[2] Singh A, Tetreault L, Kalsi-Ryan S, et al. Global prevalence and incidence of traumatic spinal cord injury[J]. Clin Epidemiol, 2014, 6:309-331.
[3] Iyer NR, Wilems TS, Sakiyama-Elbert SE. Stem cells for spinal cord injury:Strategies to inform differentiation and transplantation[J]. Biotechnol Bioeng, 2017, 114(2):245-259.
[4] Dumont RJ, Okonkwo DO, Verma S. et al. Acute spinal cord injury, part I:pathophysiologic mechanisms[J]. Clin Neuropharmacol,2001, 24(5):254-264.
[5] Tator CH. Strategies for recovery and regeneration after brain and spinal cord injury[J]. Inj Prev,2002,8(Suppl 4):V33-V36.
[6] Chen L, Li J, Wu L, et al. Synergistic actions of olomoucine and bone morphogenetic protein-4 in axonal repair after acute spinal cord contusion[J]. Neural Regen Res, 2014, 9(20):1830-1838.
[7] 任浩. 局部释放夫拉平度调控局部星形胶质细胞和炎症反应促进脊髓损伤修复[D]. 杭州:浙江大学, 2017.
[8] Ambrosius B, Pitarokoili K, Schrewe L, et al. Fingolimod attenuates experimental autoimmune neuritis and contributes to Schwann cell-mediated axonal protection[J]. J Neuroinflammat, 2017, 14(1):92.
[9] Kretzschmar B, Pellkofer H, Weber MS. The use of oral disease-modifying therapies in multiple sclerosis[J]. Curr Neurol Neurosci, 2016, 16(4):38.
[10] Chun J, Hartung HP. Mechanism of action of oral fingolimod (FTY720) in multiple sclerosis[J].Clin Neuropharmacol, 2010, 33(2):91-101.
[11] Dusaban SS, Chun J, Rosen H, et al. Sphingosine 1-phosphate receptor 3 and RhoA signaling mediate inflammatory gene expression in astrocytes[J]. J Neuroinflammat, 2017, 14(1):111.
[12] Zhu Z, Fu Y, Tian D, et al. Combination of an immune modulator fingolimod with alteplase in acute ischemic stroke:a pilot trial[J]. Circulation, 2015, 132(12):1104-1112.
[13] Zhang J,Zhang A,Sun Y, et al. Treatment with immunosuppressants FTY720 and tacrolimus promotes functional recovery after spinal cord injury in rats[J]. Tohoku J Exp Med, 2009,219(4):295-302.
[14] Norimatsu Y, Ohmori T, Kimura A, et al. FTY720 improves functional recovery after spinal cord injury by primarily nonimmunomodulatory mechanisms[J]. AmJPathol, 2012, 180(4):1625-1635.
[15] Chen L, Li J, Wu L, et al. Synergistic actions of olomoucine and bone morphogenetic protein-4 in axonal repair after acute spinal cord contusion[J]. Neural Regen Res, 2014, 9(20):1830-1838.
[16] Ren H, Han M, Zhou J, et al. Repair of spinal cord injury by inhibition of astrocyte growth and inflammatory factor synthesis through local delivery of flavopiridol in PLGA nanoparticles[J]. Biomaterials, 2014, 35(24):6585-6594.
[17] Choi JW, Gardell SE, Herr DR, et al. FTY720(fingolimod) efficacy in an animal model of multiple sclerosis requires astrocyte sphingosine 1-phosphate receptor 1(S1P1) modulation[J]. Proc Natl Acad Sci USA, 2011,108(2):751-756.
[18] Snyder EY, Teng YD. Stem cells and spinal cord repair[J]. N Engl J Med, 2012, 366(20):1940-1942.
[19] Hsuan YC, Lin C, Chang C, et al. Mesenchymal stem cell-based treatments for stroke, neural trauma, and heat stroke[J]. Brain Behav, 2016, 6(10):1-11.

芬戈莫德对神经干细胞和星形胶质细胞增殖及迁移的影响及其对小鼠脊髓损伤的保护机制

Effect of fingolimod on proliferation and migration of neural stem cells and astrocytes and its protective mechanism against spinal cord injury in mice

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 2

[1]	刘芳, 陈佳, 苏华. 不同时间窗高压氧干预对大鼠脊髓损伤后细胞凋亡及脊髓功能恢复的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 578-583.
[2]	赵岩, 肖宇龙, 左媛, 王喜良, 霍洪军, 江建明, 闫慧博. 神经干细胞移植联合促红细胞生成素对大鼠脊髓损伤的修复作用[J]. 吉林大学学报(医学版), 2015, 41(02): 333-337.
[3]	王喜良, 赵岩, 左媛, 肖宇龙, 霍洪军, 江建明, 闫慧博. 神经干细胞移植联合腹腔注射促红细胞生成素对横断性脊髓损伤大鼠神经轴突的修复作用[J]. 吉林大学学报(医学版), 2015, 41(01): 99-104.
[4]	张秀英，薛辉，李忱，刘佳梅，李岩，陈东. NDGA对移植入损伤脊髓后的化学去细胞肌肉支架中的羊膜上皮细胞存活的影响[J]. 吉林大学学报(医学版), 2013, 39(6): 1116-1120.
[5]	孙艺学，王爱兵，李子义，张学明 . 成体大脑神经发生特点及其影响因素研究进展[J]. 吉林大学学报(医学版), 2013, 39(5): 1076-1080.
[6]	刘健\|杨小玉\|夏威威\|董健\|杨茂光\|矫健航. 脊髓损伤后神经功能可塑性机制及影响因素的研究进展[J]. J4, 2012, 38(2): 386-389.
[7]	孙翊夫\|李英普\|刘鹏\|潘肃\|程连杰\|张远鹰\|高忠礼 . 急性颈脊髓损伤患者早期气管切开治疗的意义[J]. J4, 2011, 37(6): 1133-1135.
[8]	齐玲\|金宏\|丁丽娟\|温娜\|唐泽波\|狄阳\|李蕴潜. CD133免疫磁珠分选脑肿瘤干细胞及其生物学特性[J]. J4, 2011, 37(3): 441-444.
[9]	蔺宇\|刘少君. SCIRR 10截短体真核表达载体的构建及其在COS-7细胞中的表达[J]. J4, 2011, 37(2): 226-230.
[10]	李忱\|薛辉\|刘佳梅\|张秀英\|宋宇\|王金成. 羊膜上皮细胞移植对大鼠脊髓半横断损伤后神经再生的促进作用[J]. J4, 2011, 37(2): 202-206.
[11]	薛辉, 陈东, 张秀英, 刘颖. 化学去细胞肌肉组织工程支架与大鼠脊髓的生物相容性[J]. J4, 2009, 35(5): 801-804.
[12]	崔俐, 张巨, 林卫红, 马涤辉, 齐亚丽, 付海龙, 孙强, 常明, 吕晓红. 谷氨酸对体外培养胎鼠神经干细胞的损伤作用[J]. J4, 2009, 35(3): 451-455.
[13]	张殿君，刘一，付长峰，宿晓云，王芳，李相军. 新生大鼠脊髓源性神经干细胞的分离培养及分化鉴定[J]. J4, 2009, 35(2): 230-232.
[14]	李颖智, 金海鸿,沈小亮, 杨有赓 ,李志洲. 截瘫大鼠血液生化及骨密度改变[J]. J4, 2006, 32(5): 835-838.
[15]	付长峰，刘一，李相军，沈波，张绍昆，宋之明，张新. IGF-1抑制SCI大鼠脊髓前角神经元凋亡的体内实验[J]. J4, 2006, 32(4): 568-570.