摘要
目的探讨骨髓间充质干细胞移植治疗大鼠急性脊髓损伤的作用机制。
方法1.运用改良Allen氏打击器制备大鼠T10脊髓损伤模型,假手术组48只,损伤组96只随机分为对照组和移植组,各组再分为移植术后1天、3天、7天、14天4个亚组,每亚组12只大鼠。对照组和移植组于损伤后7天分别经尾静脉分别注射磷酸盐缓冲液(PBS)1ml和含转绿色荧光蛋白的大鼠骨髓基质干细胞(GFP-BMSCs)单细胞PBS悬液1ml(1x106个BMSCs)。
2.移植术后1天、3天、7天、14天,应用BBB评分法评价治疗前后损伤大鼠运动功能改善状况。
3.移植术后14天,应用光镜及电子显微镜观察损伤脊髓组织形态学变化；移植术后3天、7天,应用荧光激发光技术观察BMSCs在体内迁移、存活情况。
4.移植术后1天、3天、7天、14天,应用免疫组织化学技术检测损伤大鼠Nogo-A及SYN的表达。
结果1.移植组与对照组损伤大鼠的运动功能均有不同程度的恢复。移植组损伤区脊髓结构与对照组相比,相对较完整,表现出较明显的运动功能改善(P<0.05)。
2.移植组可见GFP标记的细胞在损伤区及其周边区明显聚集并存活。
3.移植术后第3天、第7天和第14大,移植组脊髓损伤区周边组织中Nogo-A表达较对照组明显降低(P<0.05)。
4.移植术后第3天、第7天和第14天,移植组脊髓损伤区周边组织中SYN表达较对照组明显升高(P<0.05)。
结论
1.BMSCs通过外周血管移植能够穿透血脑屏障,向脊髓受损区域发生迁徙,并在其内存活、分化促进大鼠急性脊髓损伤的神经功能的恢复。
2.BMSCs移植治疗急性脊髓损伤可抑制受损局部Nogo-A蛋白的表达,从而减少胶质瘢痕的形成,促进轴突的再生。
3.BMSCs移植治疗急性脊髓损伤有利于受损局部突触素(SYN)表达增加,促进突触功能的恢复。
Objectives To study therapeutic benefit and mechanism of intravenous administration of bone marrow mesenchymal stem cells after spinal cord injury in adult rats.
Methods .Rat's acute SCI model was established with modified Allen's method。There are 48 in the sham-operated group and 96 rats in the injuried group. Then the injuried group was randomly divided into transplantation group and control group.There were 4 time points in each group,n=12 in each point. At 7days after SCI,5×l06 BMSCs (1ml) were injected into the transplantation group by tail vein and 1ml (PBS) were given to the control group by tail vein. BMSCs were labeled by GFP before transplantation。
2.Functional out come meaurements using the Basso-Beatti-Bresneh-
an Score were performed at the 1st d、3rd d,7th d and 14th d after transplantation.
3.Light microscopy and electron microscopy were used to identify the morphology of the sections of injured spinal cord tissue at 14th d after transplantation. Laser scanning microscopy were used to identify migrati-on and survival BMSCs in vivo at the 3rd d and 7th d after transplantation.
4.The expressions of Nogo-A and SYN were detected by immunohistoc-hemical method at the 1st d、3rd d,7th d and 14th d after transplantation.
Results 1.neurological functions of transplantation group and control group all were recoveried in varying degrees,but neurological function in the transplantation group was higher than it in the control group (P＜0.05).
2.GFP staining positive cells were obviously gathered and survived in the injury zone and its surrounding area at the transplantation group.
3.the expressions of Nogo-A in the transplantation group was down-regulated more significantly than the control group at the 3rd d、7th d、14th d after transplantation.(P<0.05)
4.the expressions of SYN in the transplantation group was higher significantly than the control group at the 3rd d、7th d、14th d after transplantation.(P<0.05)
Conclusion
1.BMSCs transplanted by vein can penetrate through the blood-brain barrier, to spinal cord injury, and regional occurred in vitro migrating live, differentiation promote rat acute spinal cord injury nerve function recovery o
2.BMSCs transplantation in the treatment of acute spinal cord injury can inhibit the damaged local Nogo-A protein expression, thus reducing glial scar formation, promote the regeneration of axons.
3.Transplantation in the treatment of acute spinal cord injury BMSCs to local SYN expressing increasing, promote synaptic function recovery.
引文
1 Llorens F, Gil V, del Rio JA.. Emerging functions of myelin-associated proteins during development, neuronal plasticity, and neurodegeneration. FASEB J.2011 Feb;25(2):463-75.
2 Tatagiba M,Brosamle C,Schwab ME.Regeneration of injury axons in the adult mammalian central nervous system [J].Neurosurg,1997,40:541-547.
3 Service RF. Tissue engineers build new bone[J].Science,2000,289:1498-1500
4 Chopp M,Zhang XH,Li Y,et al.Spinal cord injury in rat treatment with bone marrow stromal cell transplantation[J].Neuroreport,2000,11(13):3001-3005.
5 Wu S,Suzuki Y,Noda T,et al. Bone marrow stromal cells enhance differentiation of cocultured neurosphere cells and promote regeneration of injured spinal cord[J].Neurosci Res,2003,72 (3):343-51.
6 ZuritaM, Vaquero J. Functional recovery in chronic paraplegia after bone marrow stromal cells transplantation [J]. Neuroreport,2004,15 (7):1105-1108.
7 Basso DM, Beattie MS, Bresnaha JC. A sensitive and reliable locomitor rating scale for open field testing in rats.[J] Neurotrauma.1995,12(1):-21.
8 Satake K, Lou J, Lenke LG. Migration of mesenchymal stem cells throughcerebrospinal fluid into injured spinal cord tissue[J].Spine,2004,29 (18):1971-1979
9赵宗茂,张庆俊,韩中朝等骨髓间质干细胞移植对大鼠脊髓损伤神经功能恢复的影响[J].中华神经外科杂志2003,1(19)：58-62
10 Li Y,Chopp M,Chen J, et al. Intrastriatal transplantation of bone marrow nonhematopoietic cells improves functional recovery after stroke in adult mice[J].Cereb Blood Flow 2000,20 (9):1311-1319.
11 Chen J, Li Y, Wang L et al. Therapeutic benefit of intracerebral transplantation of bone marrow stromal cells after cerebral ischemia in rats[J].Neurol Sci 2001,189(1-2):49-57.
12 Ide C, Nakai Y, Nakano N, Seo TB. et al. Bone marrow stromal cell transplantation for treatment of sub-acute spinal cord injury in the rat[J].Brain Res.2010 May 21;1332:32-47
13 ZuritaM, Vaquero J, Bonilla C, et al. Functional recovery of chronic paraplegic pigs after autologous transplantation of bone marrow stromal cellsf J]. TransPlantati-n,2008,86 (6):845-853.
14 Mahmood A,Lu D,Li Y,et al.Intracranial bone marrow transplantation after traumatic brain injury improving functional outcome in adult rats[J]. Neurosurg,2001,94:589-595.
15 Lu D, Mahmood A, Wang L, et al. Adult bone marrow stromal cells administered intravenously to rats after traumatic brain injury migrate into brain and improve neurological outcome[J].Neuroreport 2001,12(3):559-563.
16 RomboutsWJ, Ploemacher RE. Primary murine MSC show highly efficient homing to the bonemarrow but lose homing ability following culture [J]. L eukemia,2003,17 (1):160-170
17 Karina T. Wright,Wagih El Masri,Aheed Osman,et al. Bone Marrow for the Treatment of Spinal Cord Injury:Mechanisms and Clinical Applications.STEM CELLS 2011;29:169-178
18 W ang L,L i Y, Chen J, et al. MCP21,M IP2I, IL 28 and ischemic cerebral tissue enhance human bone marrowstromal cellmigration in interface culture [J]. Hemato logy, 2002,7(2):113-117.
19. Woolf CJ, Bloechlinger S. It takesmore than two to Nogo. Science,2002; 297: 1132-1134
2O.GrandPreT, Nakamura F, Vartanian T et al. Identification of the Nogo inhibitor of axon regeneration as a Reticulon protein. Nature,2000; 403:439-444
21. ChenMS, HuberAB, SchwabME et al. Nogo-A is amyelin-associated neurite outgrowth inhibitor and an antigen formonoclonal anti-body IN-1. Nature,2000; 403: 434-439
22. Prinjha R, Moore SE, VinsonM et al. Inhibitor of neurite outgrowth in humans. Nature,2000; 403:383-384
23. Josephson A, T rifunovski A, Widmer HR, et al. Nogo-recept orgeneact-ivity: cellular localization and development al regulat ion of mRNA in mice and humans [J]. J Comp Neurol,2002,453(3):292
24 Hunt D, Mason M, Campebell G, et al. Nogo recept or mRNA expression in int act and regenerat ing CNS neurons [J]. Mol Cell Neurosci,2002,20 (1153):537.
25杨磊,周长满,鄂玲玲,等Nogo(N-18)在正常大鼠脊髓和坐骨神经的分布及损伤后变化的免疫组织化学研究[J].解剖学报,2002,33(4)：370.
26. NgCE, TangBL, TangBL. Nogos and Nogo-66 receptor:factors inhibiting CNS neuron regeneration. Neurosci Res,2002; 67:559-565
27.Caroni P,Schwab ME.Two membrane protein fractions from rat central myelin with inhibitory properties for neurite growth and fibroblast spreading.J Cell Biol.1988 Apr;106(4):1281-1288.
28. Caroni P, Schwab ME. Antibody against myelin-associated inhibitor of neurite growth neutralizes nonpermissive substrate properties of CNS white matter. Neuron 1998,1:85-96.
29. Raineteau O,fouad K,bareyre FK, et al.Reorganization of descending motor tracts in the rat spinal cord.Eor J Neurosci,2002,16.(9).1761-1771.
30. Wiessner C,Bareyre FM,Allegreini PR,et al.Anti-nogo-antibody infusion 24 hour after experimental stroke improved behavioral outcome and corticospinal piasticity in normotensive and spontaneously hypertensive rats.J Cereb Blood Flow Metab,2003,23(2); 154-165.
31 GrandPre T, Shuxin Li,Strittmatter SM.Nogo-66 receptor antagonist peptide promotes axonal regeneration.Nature,2002,417:547-551.
32. Sun AP, Zhang J, Liao Q, et al. Significance of Nogo-A expression of atrophic muscle fibers in diagnosis of amyotrophic lateral sclerosis. Beijing Da Xue Xue Bao.2011 Apr 18;43(2):238-241.
33 Son BR, Marquez-Curtis LA, Kucia M, et al. Migration of bone marrow and cord blood mesenchymal stem cells in vitro is regulated by stromal -derived factor-1-CXCR4 and hepatocyte growth factor-c-met axes and involves matrix metalloproteinases[J]. Stem Cells,2006,24:1254—1264.
34 Belien AT, Paganetti PA, Schwab ME. Membrane type 1 matrix metalloprotease (MT1-MMP) enables invasive migration of glioma cells in central nervous system white matter[J]. Cell.Biol,1999,144:373—384.
35 Wright KT, El Masri W, Osman A, et al. Bone marrow stromal cells stimulate neurite outgrowth over neural proteoglycans (CSPG), myelin associated glycoprotein and Nogo-A [J]. Biochemical and Biophysical Research Communications,2007, 354:559—566.
36 Janh R,Schiebler W,Quimet C, et al. A238000 Dalton membrane protein(p38) present in synaptic vesicle [J]. Proc Natl Acad Sci US A,1985,82 (12):4137-4141.
37 Janz R,Sudhof TC,Hammer RE, et al. Essention roles in synaptic plascity for synaptogyrin I and synaptophysin [J]. Neuron,1999,24 (3):687-700.
38 Tarsa L, Goda Y. Synaptophysin regulates activity2dependent synase formation in cultured hippocampal neurons [J]. Proc Natl Acad Sci USA,2002,99 (2):1012-1016.
39黄如训,张艳,方燕南,等.大鼠脑梗死后运动功能可塑性物质基础及发生机制[J].中华医学杂志,2000,80(10)：769-772.
40 Mullany PM,Lynch MA. Evidence for a role for synatophysin in expression of long-term Potentiation in rat dentate gyrus [J]. Neuroreport,1998,9 (1):2489.
41 Sufan Wu,Yoshihisa Suzuki,Yoko Ejiri,etal,Bonemarrow stromal cells enhance differentitati-on of cocultured neurosphere cells and promote regeneration of injured spinal cord[J].Neuroscie-nce Research 2003,72:343-351.
42 Mahmood A,Lu D,Li Y,et al.Intracranial bone marrow transplantation after traumatic brain injury improving functional outcome in adult rats[J].Neurosurg,2001, 94:589-595.
43 Chen X, Katakowdki M, Li Y,et al.Human bone marrow stromal cell cultures conditioned by traumatic brain tissue extracts:growth factor production[J].Neurosci Res, 2002,69 (5):687-91.
1 Kwon BK, F isher CG, Dvo rak M F, et al. Strategies to promote neural repair and regeneration after spinal cord injury. Spine,2005,30 (17 Supp 1):3-13.
2 P ierdomenico L, Bonsi L, Calvitti M, et al. M ultipo tentmesenchymal stem cells w ith immuno supp ressive activity can beeasily isolated from dental pulp. T ransp lantation,2005,80 (6):836-842.
3韩冰,付小兵.间充质干细胞的研究进展与临床应用前景.中国修复重建外科杂志,2006,20(12)：1257-1261.
4 Foster LJ, Zeemann PA, L i C, et al. D ifferential expression profiling of membrane pro-teins by quantitative proteomics in ahuman mesenchymal stem cell line undergoing osteoblast differentiation. Stem Cells,2005,23 (9):1367-1377.
5 M ajumdar M K, Keane-Moore M, Buyaner D, et al.Characterization and functionality of cell surface mo lecules onhuman mesenchymal stem cells. J Biomed Sci, 2003,10 (2):228-241.
6 Silva WA Jr, CovasDT, PanepucciRA, et al. The profile of gene expression of human marrow mesenchymal stem cells. Stem Cells,2003,21 (6):661-669.
7 W islet-Gendebien S, Hans G, L ep rince P, et al. P lasticity ofcultured mesenchymal stem cells:sw itch from nestin-positive toexcitable neuron-like pheno type. Stem Cells, 2005,23 (3):392-402.
8 Yoon YS, W ecker A, Heyd L, et al. Clonally expanded novelmultipo tent stem cells from human bone marrow regeneratemyocardium after myocardial infarction. J Clin Invest,2005,115(2):326-338.
9 晏杰, 刘玲蓉, 张其清. 诱导骨髓间充质干细胞向软骨分化的体外研究.中国修复重建外科杂志,2006,20(11)：1114-1118.
10韩冰,付小兵,孙晓庆.骨髓基质细胞基因表达的研究进展.中国修复重建外科杂志,2005,19(2)：153-158.
11董世武,应大军,朱楚洪,等.核心结合因子A1基因修饰骨髓间充质干细胞对骨缺损的修复研究.中国修复重建外科杂志,2006,20(5)：555-559.
12 AllersC, SierraltaWD, Neubauer S, et al. Dynamic of distribution of humanbone marrow2derived mesenchymal stem cells after transplantation into adult unconditioned mice[J]. Transplantation,2004,78 (4):503-508.
13 Furuya T, HashimotoM, KodaM, et al. Treatment of rat spinal cord injurywith a Rho2kinase inhibitor and bone marrow stromal cell transplantation[J]. Brain Res,2009, 1295:192-202.
14 Nishida K, TanakaN, Nakanishi K, et al.Magnetic targeting of bonemarrow stromal cells into spinal cord:through cerebrospinal fluid[J]. Neurore2port,2006,17(12):1269 1272.
15 Satake K, Lou J, Lenke LG. Migration of mesenchymal stem cells throughcerebrospinal fluid into injured spinal cord tissue[J].Spine,2004,29 (18):1971-1979
16 ZuritaM, Vaquero J, Bonilla C, et al. Functional recovery of chronic paraplegic pigs after autologous transplantation of bone marrow stromal cells[J]. Transplantation,2008, 86 (6):845-853.
17 Ide C, Nakai Y, Nakano N, Seo TB, et al. Bone marrow stromal cell transplantation for treatment of sub-acute spinal cord injury in the rat[J j.Brain Res.2010 May 21;1332:32-47
18 RonsynMW, Daans J, Spaepen G, et al. Plasmid-based genetic modification of human bone marrow-derived stromal cells:analysis of cell survivaland transgene expression after transplantation in rat spinal cord [J]. BMCBiotech,2007,7:90.
19 Ding Y, Yan Q, Ruan JW, et al. Electro-acupuncture promotes survival,differentiation of the bone marrow mesenchymal stem cells aswell as func2tional recovery in the spinal cord2transected rats[J]. BMC Neurosci,2009,10:35.
20 HwangDH, Kim BG, Kim EJ, et al. Transplantation of human neural stemcells transduced with Olig2 transcription factor improves locomotor recoveryand enhancesmyelination in the white matter of rat spinal cord followingcontusive injury[J]. BMC Neurosci,2009,10:117.
21马希峰,杨波.神经干细胞移植治疗大鼠脊髓损伤[J].郑州大学学报,2003,38(1)：21-23.
22 O kanao H,O gaw a Y,N akamuraM, et al. T ransplantation of neural stem cells into the sp inal cord after injury [J]. Sem inars Cell Developmental B io logy,2003,14: 191-198.
23 Hofstetter CP, Schwarz EJ, Hess D, et al. Marrow stromal cells from guiding strands in the injured spinal cord and promote recovery. P roc N atlA cad Sci USA,2002, 99 (4):2199-2204.
24 Satake K, Lou J, L enke L G. Migration of mesenchymal stem cells th rough cerebro sp-inal fluid into injured spinal cord tissue.Spine,2004,29 (18):1971-1979.
25 A nkeny DP, M cT ique DM, J akeman LB, et al. Bone marrow transplants provide tissue proection and directional guidance for axon after contusive spinal cord injury in rats. Exp N euro 1,2004,190 (1):17-31.
26 Okano H, Ogawa Y, N akamura M, et al. Transplantation of neural stem cell into the sp-inal cord after injury. Sem in Cell Dev Bio,2003,14 (3):191-198.
27 Inoue M, Honmou O, Oka S, et al. Comparative anylasis of remyelinating potential of focal and intravenous administration of auto logous bone marrow cells into the rat demyelinated spinal cord. Glia,2003,44 (2):111-118.
28 Anjos-A fonso F, Siapati EK,Bonnet D, et al. In vivocontribution of murine mesenchymal stem cells intomultiple cell-types under m inimal damage condit ions[J]. J Cell Sci,2004,117 (23):5655-5664.
29 Hofstet ter CP, Schw arz EJ, Hess D, et al. M arrowstromal cells fo rm guiding strands in the injured spinal cord and promote recovery[J]. P roc N at 1A cadSciU SA, 2002,99 (4):2199-2204.
30 Zhang J,L i Y, Chopp M, et al. Exp ression of insulin-like grow th factor 1 and receptor in ischemic ratst reated w ith human marrow stromal cells [J]. B rainRes,2004, 1030(1):19-27.
31 BoidoM, Rupa R, GarbossaD, et al. Embryonic and adult stem cells promote raphespinal axon outgrowth and improve functional outcome following spinal hemisection in mice[J]. Eur J Neurosci,2009,30 (5):833-846.
32 RomboutsWJ, Ploemacher RE. Primary murine MSC show highly efficient homing to the bonemarrow but lose homing ability following culture [J]. L eukemia,2003, 17(1):160-170
33 Karina T. Wright.Wagih El Masri,Aheed Osman,et al. Bone Marrow for the Treatment of Spinal Cord Injury:Mechanisms and Clinical Applications.STEM CELLS 2011;29:169-178
34 W ang L,L i Y, Chen J, et al. MCP21.MIP21. IL 28 and ischemic cerebral tissue enhance human bone marrowstromal cellmigration in interface culture [J]. Hemato logy, 2002,7(2):113-117.
35 Zurita M,V aquero J. Bone marrow stromal cells can achieve cure of chronic paraplegic rats:functional and morphological outcome one year after transplantation [J]. Neuro science Letters,2006,402 (10):51-56.
36 Chen J, Zhang ZG, L i Y, et al. Int ravenous adm inist rat ion of human bone marrow st romal cells induces angiogenesis in the ischem ic boundary zone afterst roke in rats[J]. Circ Res,2003,92 (6):692-699.
37 Zhang ZG, Zhang L, J iang Q, et al. Bone marrow-derived endo thelial p rogenito r cells part icipate in cerebral neovascularizat ion after focal cerebral ischemiain the adult mouse [J]. Circ Res,2002,90 (3):284-288.
38 Hofstetter CP, Schwarz EJ, Hess D, et al. M arrow stromal cellsfrom guiding strands in the injured sp inal co rd and p romo terecovery. P roc N atlA cad SciU SA,2002, 99 (4):2199-2204.
39 Satake K, Lou J, L enke L G. Migration of mesenchymal stemcells th rough cerebro spinal fluid into injured spinal cord tissue.Spine,2004,29 (18):1971-1979.
40 Ankeny DP, M cT ique DM, J akeman LB, et al. Bone marrowtransplants provide tissue p-rotection and directional guidance fo raxon after contusive spinal cord injury in rats. Exp N euro l,2004,190 (1):17-31.
1. Llorens F, Gil V, del Rio JA.. Emerging functions of myelin-associated proteins during development, neuronal plasticity, and neurodegeneration. FASEB J.2011 Feb;25(2):463-75.
2. GrandPreT, Nakamura F, Vartanian T et al. Identification of the Nogo inhibitor of axon regeneration as a Reticulon protein. Nature,2000; 403:439-444
3. ChenMS, HuberAB, SchwabME et al. Nogo-A is amyelin-associated neurite outgrowth inhibitor and an antigen formonoclonal anti-body IN-1. Nature,2000; 403: 434-439
4. Prinjha R, Moore SE, VinsonM et al. Inhibitor of neurite outgrowth in humans. Nature,2000; 403:383-384
5.HuberAB, Weinmann O, Brasamle C et al. Patterns of Nogo mRNA and protein exp-ression in the developing and adult rat and after CNS lesions. J Neurosci,2002; 22: 3553-3567
6.HuntD, Mason MR, Campbell G et al. Nogo receptor mRNA expression in intact and regenerating CNS neurons. Mol Cell Neurosci,2002; 20:572-552
7. Liu H, Ng CE, Tang BL et al. Nogo-A expression in mouse centralnervous system neurons. Neurosci Lett,2002; 328:257-260
8. HuntD, Coffin RS, Prinjha RK et al. Nogo-A exp ression in the intact and injured nervous system. Mol CellNeurosci,2003; 24:1083-1102
9. Chen Y, Tang X, Cao X, et al. Human Nogo-C overexpression induces HEK293 cell apoptosis via a mechanism that involves JNK-c-Jun pathway [J]. Biochem Biophys ResCommun (S0006-291X),2006,348 (3):923-928
10. Josephson A, T rifunovski A, Widmer HR, et al. Nogo-recept orgeneact-ivity: cellular localization and development al regulat ion of mRNA in mice and humans [J]. J Comp Neurol,2002,453(3):292.
11. Hunt D, Mason M, Campebell G, et al. Nogo recept or mRNA expression in int act and regenerat ing CNS neurons [J]. Mol Cell Neurosci,2002,20 (1153):537.
12.杨磊,周长满,鄂玲玲,等.Nogo(N-18)在正常大鼠脊髓和坐骨神经的分布及损伤后变化的免疫组织化学研究[J].解剖学报,2002,33(4)：370.
13. Fournier AE, GrandPre T,Strittmatter SM. Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration[J]. Nature,2001,409 (6818):341-346.
14.范有名.NgR受体复合物介导中枢神经再生抑制机制研究[J].华南国防医学杂志,2010,24(1)：72-74.
15. Fournier AE, Gould GC, Liu BP, et al. Truncated soluble Nogo recepter binds Nogo-66 inhibition of axon growth by myelin[J] J Eurosci,2002,22(20):887 6,
16. Ng CE, T ang BL. Nogos and the Nogo-66 receptor:factors inDhibiting CNS neuron regeneration[J]. J Neurosci Res,2002,67(5):559.
17. FournierAE, Grandpre T, Strittmatter SM et al. Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration. Nature,2001; 409:341-346
18. Josephon A, Widenfalk J, Widmer HW et al. Nogo mRNA expression in adult and fetal human and rat nervous tissue and in weight drop injury. Exp Neurol,2001; 169: 319-328
19.谢林,贺翔鸽,何凤慈等.Nogo266受体在大鼠神经组织中的表达.第三军医大学学报,2005：27：2335-2337
20. NgCE, TangBL, TangBL. Nogos and Nogo-66 receptor:factors inhibiting CNS neuron regeneration. Neurosci Res,2002; 67:559-565
21. Park JB, Yiu G, Kaneko S, et al. A TNF recep tor familymember,TROY, is a coreceptor with Nogo receptor in mediating the inhibitoryactivity of myelin inhibitors[J]. N euron,2005,45(3):345-351.
22.Wang K C, Kim J A, Sivasankaran R, et al. P75 interactswith the Nogo receptor as a coreceptor for Nogo, MAG and OMgp [J]. Nature,2002,420:74～78.
23. Satoh J, Tabunoki H, Yamamura T et al. TROY and LINGO-1 expression in astrocytes and macrophages/microglia in multiple sclerosis lesions. Neuropathol App 1 Neurobiol,2007; 33:99-107
24. Mi S, Lee XH, Shao ZH, et al. LINGO 1 is a component of the Nogo 66 receptor/p75 signaling complex[J].Nat Neurosci,2004,7 (3):221-228.
25. Yamagishi S, Fujitani M, Hata K, et al.Wallerian degeneration involves Rho/Rho2kinase signaling[J].J Biol Chem,2005,280 (21):20384-20388.
26. Yamashita T, Tohyama M. The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI [J].Nat Neurosci,2003,6 (5):461-467.
27.Caroni P,Schwab ME.Two membrane protein fractions from rat central myelin with inhibitory properties for neurite growth and fibroblast spreading.J Cell Biol.1988 Apr;106(4):1281-1288.
28. Caroni P, Schwab ME. Antibody against myelin-associated inhibitor of neurite growth neutralizes nonpermissive substrate properties of CNS white matter. Neuron 1998,1:85-96.
29.Freund P, Schmidlin E, Wannier T, et al. Anti-Nogo-A antibody treatment promotes recovery of manual dexterity after unilateral cervical lesion in adult primates--re-examination and extension of behavioral data[J]. Eur J Neurosci,2009,29(5):983—996.
30.Raineteau O,fouad K,bareyre FK, et al.Reorganization of descending motor tracts in the rat spinal cord.Eor J Neurosci,2002,16.(9).1761-1771.
31. Wiessner C.Bareyre FM,Allegreini PR,et al.Anti-nogo-antibody infusion 24 hour after experimental stroke improved behavioral outcome and corticospinal piasticity in normotensive and spontaneously hypertensive rats.J Cereb Blood Flow Metab,2003,23(2);154-165.
32. GrandPre T, Shuxin Li.Strittmatter SM.Nogo-66 receptor antagonist peptide promotes axonal regeneration.Nature,2002,417:547-551.
33. Tozaki H, Kawasaki T, Takagi Y, et al.Expression of nogo protein by growing axons in the developing nervous system. Brain Res Mol Brain Res,2002,104(2): 111-119.
34. Kim J E, Li S, Grandpre T, et al. Axon regeneration in young adult mice lacking Nogo-A/B [J]. Neuron,2003,38 (2):187-199.
35. Fiedler M, Hom C, Bandtlow C, et al. An engineered IN-1 F(ab) fragment with improved affinity for the Nogo-A axonal growth inhibitor perm-its immunochemical deterction and shows enhanced neutraliziing activity.Protein Eng,2002,15(11):931-941.
36. Sun AP, Zhang J, Liao Q, et al. Significance of Nogo-A expression of atrophic muscle fibers in diagnosis of amyotrophic lateral sclerosis. Beijing Da Xue Xue Bao.2011 Apr 18;43(2):238-241.
