Nogo-A及Nogo受体拮抗剂在新生大鼠缺血缺氧性脑损伤中的作用研究
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摘要
目的围产期窒息所致的新生儿缺氧缺血性脑病(hypoxic-ischemicencephalopathy,HIE)是严重威胁新生儿生命和健康的常见病之一,死亡率和致残疾率高,关键原因在于中枢神经损伤后缺乏再生的能力,一般对症治疗不能有效改善其症状及预后。传统的观点认为中枢神经损伤后不能再生,而近年的研究发现阻止中枢神经损伤后再生的重要原因是中枢神经的内环境中存在抑制神经再生的因子,目前已确认的中枢神经髓鞘来源的抑制因子至少有以下三种:Nogo-A、髓磷脂相关糖蛋白(myelin-associated glycoprotein,MAG)和少突胶质细胞-髓磷脂糖蛋白(oligodendrocyte-myelin glycoprotein,OMgp),其中Nogo-A是其中抑制作用最强的一种,其羧基端两个跨膜区之间存在一个66个氨基酸的结构域(Nogo-66),这三种抑制因子通过相同的受体(Nogo receptor,NgR)发挥抑制神经再生的作用。NEP1-40是针对Nogo-66氨基序列设计的NgR竞争性拮抗剂,能阻断中枢神经抑制因子与NgR的结合。神经节苷脂为近年常用于治疗HIE的药物,有维持细胞内外离子平衡、防止细胞内Ca~(2+)积聚及对抗兴奋性氨基酸的神经毒性等作用。本研究通过制备新生大鼠HIBD标准模型,对HIBD后大鼠分别腹腔注射NEP1-40及单唾液酸四己糖神经节苷脂钠(GM-1),观察各组大鼠脑重的增长、左右脑重差值、脑组织病理变化及Nogo-A mRNA的表达,探讨Nogo-A在新生大鼠HIBD中的作用,并观察NEP1-40与GM-1对大鼠Nogo-A表达的影响及对神经损伤后的保护作用,为进一步研究新生儿缺氧缺血性脑病有效治疗方式提供动物实验依据。
方法将100只7日龄新生大鼠随机分为空白对照组、假手术组、缺氧缺血性脑损伤(HIBD)模型组、NEP1-40治疗组及GM-1治疗组,以上各组均在6h、24h两时间点采集标本,每组10只。HIBD组及治疗组参照Rice法制备标准HIBD模型,治疗组于造模后即刻腹腔注射NEP1-4010mg/kg、GM-120mg/kg,空白对照组、假手术组与模型组腹腔注射生理盐水0.25ml/kg。各组动物分别处理后于6h、24h两时间点断颈处死。将脑组织标本固定于30%蔗糖、4%多聚甲醛溶液中静置72h,待下沉后进行冰冻切片,HE染色观察各组大鼠脑组织病理变化,用原位杂交的方法观察Nogo-A mRNA的表达,40倍镜下计数大鼠脑组织Nogo-A mRNA阳性细胞数目。
结果①HIBD组两时段,Nogo-mRNA表达均高于同时段空白组及假手术组:HE染色示脑组织神经元水肿、尼氏小体溶解坏死,脑实质内有散在的凝固性坏死,脉络丛上皮局部缺失。②NEP1-40治疗组能抑制HIBD后mRNA的表达,与同时段模型组对比有显著统计学意义;神经元轻度水肿,未见溶解坏死的尼氏小体,脉络丛上皮部分缺失,毛细血管基本正常。③神经节苷脂治疗组mRNA的表达在6h时与HIBD组无明显差异,在24h时较HIBD组低但高于空白组。24h神经元细胞水肿较模型组减轻,可见少量坏死尼氏小体,脉络丛上皮连接基本正常。
结论Nogo-A mRNA在缺氧缺血性脑损伤后表达显著增高,其编码的Nogo-A可抑制中枢神经损伤后的再生,NEP1-40能拮抗这一作用,能减轻细胞水肿、改善局部血流供应,从而可能在促进缺氧缺血性脑损伤后神经再生中起到重要作用。神经节苷脂GM-1在缺氧缺血性脑损伤后也可部分抑制Nogo-A mRNA的表达,有稳定细胞膜、减轻细胞水肿的作用。
Objective The hypoxic-ischemic encephalopathy(HIE)caused by asphyxia in peripartum is a serious disease in newborn infants,with a high disability rate and mortality rate.The most important question is that the center nerve system was not able to regenerate after injury.The traditional symptomatic treatment can not improve the prognosis effectively.But lots of studies in recent years discovered that there were factors caused the failure of regeneration.At present,there were at least three factors were found in center neural myelin sheath to produce marked effects:Nogo-A,myelin-associated glycoprotein(MAG), oligodendrocyte-myelin glycoprotein(OMgp).Nogo-A was able to produce a most important effect.There was a structural domain composed by 66 amino acids in carboxyl terminus.All the three inhibiting factors effected by combined with nogo receptor.NEP1-40 was designed to aim directly at the structural domain,and it can antagonize the combination competitively. Ganglioside was commonly used to treat HIBD,it can stabilize the cellular membrane,ease cellular edema,prevent the accumulation of Ca~(2+), antagonize the neurotoxicity of excitatory amino acids.In this study, HIBD model of newborn rats were prepared,then NEP1-40 and GM-1 were injected in their belly cavity respectively.After got the brain tissue, the weights of brains,the pathological change,the contents of Nogo-A mRNA were observed to investigate the function of Nogo-A,the effects of NEP1-40 and GM-1 in the newborn rats with hypoxic ischemic brain damage (HIBD).
Methods 100 rats were randomly divided into 10 groups:normal control group,sham operated group,HIBD model group,NEP1-40 treatment group and GM-1 treatment group after HIBD at 6h and 24h.HIBD model groups were prepared by Rice method,NEP1-40(10mg/kg)and GM-1(20mg/kg)were injected in the belly cavity of two treatment groups respectively,while,normal sodium(0.25ml/kg)were injected in the belly cavity of the normal control groups,sham operated groups,and HIBD model groups.The tissues were fixed in 30%sugar solution with 4%polyoxymethylene.Then the tissues were prepared by the frozen microtome section.The pathological changes were observed by HE drum dyeing,and the contents of Nogo-A mRNA were detected by the method of in situ hybridization.
Results①The expression of Nogo-A mRNA in HIBD groups was higher than that in control groups at both time points.In the HE section,The nerve cells were edematous,Nissl's bodies were necrotic,and the endothelial cells of the choroid plexus were not complete.②In NEP1-40 groups,the expression of Nogo-A mRNA were lower than that in HIBD groups at both time points.The nerve cells were edematus lightly,and the endothelial cells and capillaries of the choroid plexus were mostly normal.③There was no great difference between the GM-1 group and HIBD group when they were treated at the 6th hour.And for the 24th-hour-group,the expression in GM-1 group was lower than that of HIBD group but higher than that of normal control group.The cellular edema was eased than in HIBD model groups, and the choroid plexus were incomplete partly.
Conclusion Nogo-A mRNA increased obviously in the newborn rats with HIBD. Nogo-A encoded by the mRNA can inhibit the regeneration of central nerve after being injured,and NEP1-40 can antagonize the function,and be able to ease cellular edema,to improve the regional flow supply of the injured nerve,thereby it can promote the regeneration.GM-1 was also able to antagonize the expression of mRNA,to stabilize the cellular membrane, to ease cellular edema of the injured nerve and to accelerate the regeneration of new nerve.
方法将100只7日龄新生大鼠随机分为空白对照组、假手术组、缺氧缺血性脑损伤(HIBD)模型组、NEP1-40治疗组及GM-1治疗组,以上各组均在6h、24h两时间点采集标本,每组10只。HIBD组及治疗组参照Rice法制备标准HIBD模型,治疗组于造模后即刻腹腔注射NEP1-4010mg/kg、GM-120mg/kg,空白对照组、假手术组与模型组腹腔注射生理盐水0.25ml/kg。各组动物分别处理后于6h、24h两时间点断颈处死。将脑组织标本固定于30%蔗糖、4%多聚甲醛溶液中静置72h,待下沉后进行冰冻切片,HE染色观察各组大鼠脑组织病理变化,用原位杂交的方法观察Nogo-A mRNA的表达,40倍镜下计数大鼠脑组织Nogo-A mRNA阳性细胞数目。
结果①HIBD组两时段,Nogo-mRNA表达均高于同时段空白组及假手术组:HE染色示脑组织神经元水肿、尼氏小体溶解坏死,脑实质内有散在的凝固性坏死,脉络丛上皮局部缺失。②NEP1-40治疗组能抑制HIBD后mRNA的表达,与同时段模型组对比有显著统计学意义;神经元轻度水肿,未见溶解坏死的尼氏小体,脉络丛上皮部分缺失,毛细血管基本正常。③神经节苷脂治疗组mRNA的表达在6h时与HIBD组无明显差异,在24h时较HIBD组低但高于空白组。24h神经元细胞水肿较模型组减轻,可见少量坏死尼氏小体,脉络丛上皮连接基本正常。
结论Nogo-A mRNA在缺氧缺血性脑损伤后表达显著增高,其编码的Nogo-A可抑制中枢神经损伤后的再生,NEP1-40能拮抗这一作用,能减轻细胞水肿、改善局部血流供应,从而可能在促进缺氧缺血性脑损伤后神经再生中起到重要作用。神经节苷脂GM-1在缺氧缺血性脑损伤后也可部分抑制Nogo-A mRNA的表达,有稳定细胞膜、减轻细胞水肿的作用。
Objective The hypoxic-ischemic encephalopathy(HIE)caused by asphyxia in peripartum is a serious disease in newborn infants,with a high disability rate and mortality rate.The most important question is that the center nerve system was not able to regenerate after injury.The traditional symptomatic treatment can not improve the prognosis effectively.But lots of studies in recent years discovered that there were factors caused the failure of regeneration.At present,there were at least three factors were found in center neural myelin sheath to produce marked effects:Nogo-A,myelin-associated glycoprotein(MAG), oligodendrocyte-myelin glycoprotein(OMgp).Nogo-A was able to produce a most important effect.There was a structural domain composed by 66 amino acids in carboxyl terminus.All the three inhibiting factors effected by combined with nogo receptor.NEP1-40 was designed to aim directly at the structural domain,and it can antagonize the combination competitively. Ganglioside was commonly used to treat HIBD,it can stabilize the cellular membrane,ease cellular edema,prevent the accumulation of Ca~(2+), antagonize the neurotoxicity of excitatory amino acids.In this study, HIBD model of newborn rats were prepared,then NEP1-40 and GM-1 were injected in their belly cavity respectively.After got the brain tissue, the weights of brains,the pathological change,the contents of Nogo-A mRNA were observed to investigate the function of Nogo-A,the effects of NEP1-40 and GM-1 in the newborn rats with hypoxic ischemic brain damage (HIBD).
Methods 100 rats were randomly divided into 10 groups:normal control group,sham operated group,HIBD model group,NEP1-40 treatment group and GM-1 treatment group after HIBD at 6h and 24h.HIBD model groups were prepared by Rice method,NEP1-40(10mg/kg)and GM-1(20mg/kg)were injected in the belly cavity of two treatment groups respectively,while,normal sodium(0.25ml/kg)were injected in the belly cavity of the normal control groups,sham operated groups,and HIBD model groups.The tissues were fixed in 30%sugar solution with 4%polyoxymethylene.Then the tissues were prepared by the frozen microtome section.The pathological changes were observed by HE drum dyeing,and the contents of Nogo-A mRNA were detected by the method of in situ hybridization.
Results①The expression of Nogo-A mRNA in HIBD groups was higher than that in control groups at both time points.In the HE section,The nerve cells were edematous,Nissl's bodies were necrotic,and the endothelial cells of the choroid plexus were not complete.②In NEP1-40 groups,the expression of Nogo-A mRNA were lower than that in HIBD groups at both time points.The nerve cells were edematus lightly,and the endothelial cells and capillaries of the choroid plexus were mostly normal.③There was no great difference between the GM-1 group and HIBD group when they were treated at the 6th hour.And for the 24th-hour-group,the expression in GM-1 group was lower than that of HIBD group but higher than that of normal control group.The cellular edema was eased than in HIBD model groups, and the choroid plexus were incomplete partly.
Conclusion Nogo-A mRNA increased obviously in the newborn rats with HIBD. Nogo-A encoded by the mRNA can inhibit the regeneration of central nerve after being injured,and NEP1-40 can antagonize the function,and be able to ease cellular edema,to improve the regional flow supply of the injured nerve,thereby it can promote the regeneration.GM-1 was also able to antagonize the expression of mRNA,to stabilize the cellular membrane, to ease cellular edema of the injured nerve and to accelerate the regeneration of new nerve.
引文
[1]邵肖梅.新生儿缺氧缺血性脑病的发病机理及治疗[J].中华儿科杂志,1997,35(7):289-291.
[2]周伟.缺氧缺血性脑损伤的分子生物学进展[J].国外医学儿科学分册,1998,25(1):32-35.
[3]李炜如.兴奋性氨基酸及钙离子内流在缺氧缺血性脑损伤中的作用[J].中国实用儿科学杂志,1997,12(4):245-246.
[4]王冰,周文强,周从乐.新生鼠缺氧缺血后脑组织超氧化物歧化酶活性变化[J].新生儿科杂志.2002,17(1):12-13.
[5]Fellman V,Riavio KO.Reperfusion injury as the ischemic cerebral cell death[J].Prog Cardiovasc Dis,1999,42(3):185.
[6]Goddard DR,Berry M,Kirvell SL,et al.Fibroblast growth factor-2induces astroglial and microglial reaction in vivo[J].J Anat,2002,200(3):57-67.
[7]Zaremba J.Contribution of tumor necrosis factor alpha to the pathogenesis of stoke[J].Folia Morphol(warsz),2000,59(3):137-143.
[8]欧阳小琳.新生儿缺氧缺血性脑病血液酸碱失调分析及处理[J].中国实用儿科杂志,1999,14(5):297-298.
[9]薛辛东,杜立中主编.儿科学,北京:人民卫生出版社,2005:121-122.
[10]张少丹,钱培德,张淑琴.纳洛酮治疗新生儿缺氧缺血性脑损伤的临床研究[J].中国实用儿科杂志,1996,11(4):207-208.
[11]韩玉昆.脑细胞代谢激活剂在新生儿缺氧缺血性脑病治疗中的评价[J].中国实用儿科杂志,1998,13(2):80-82.
[12]张伟,吴婉芳,李松等.高压氧治疗新生儿缺氧缺血性性脑病疗效及毒副作用观察[J].中华儿科杂志,1997,12(1):13.
[13]陈志刚,朱诚,张光霁等神经节苷脂GM-1对急性颅脑损伤的早期保护作用[J].中华神经外科杂志,1998,1(14):213-215.
[14]Rowntree S,Kolb B.Blockade of basic fibroblast growth factor retards recovery from motor cortex injury in rats[J].EurJ Neutosci,1997,9:2432-2441.
[15]David S,Aguayo AJ.Axonal elongation into peripheral nervous system bridges after central nervous system injury in adult rats[J].Science,1981,214:931-933.
[16]Yiu G,He Z.Signaling mechanisms of the melin inhibitors of axon regeneratoion[J].Curr Opin Neurobiol,2003,13:1-7.
[17]Liu BP,Fournier A,Grandpre T,et al.Myelin-associalted glycoprotein asa functional ligand for the Nogo-66 receptor[J].Science,2002,297(5584):1190-1193.
[18]Caroni P,Schwab ME.Two membrane Protein factions from rat central myelin with inhibitory properties for neurite grouth and fibroblast spreading[J].Cell Biol,1988,106(4):1281-1284.
[19]Huber AB,Schwab ME.Nogo-A,a potent inhibitor of neurite outgrouth and regeneration[J].Biol Chem,2000,381:407-419.
[20]Perry AB,Flanagan JG.Uogo domains and a Nogo receptor implications for axonregeneration[J].Neuron,2001,30:11-14.
[21]陈益存,陆东东,张锡然.RTNs家族研究进展[J].细胞生物学杂志,2005,27:53-56.
[22]Grandpre T,Wakamura·F,Vartaninan T et al.Identification of the Nogo inhibitor of axon regeneration as a reticulon protein[J].Nature,2000,403:439-443.
[23]Oertle T,Van der Haar ME,Bandtlow CE et al.Nogo-A inhibits neurite outgrowth and cell spreading with three discrete regions[J].Neurosci,2003,23(13):5393-5406.
[24]Jin WL,Lin YY,Lin HL et al.Intraneuronal location of Nogo-A in the rat[J].Comp Neurol,2003,458:1-10.
[25]Prinjha R,Morrow SE,Vinson M et al.Inhibitor of neurite outgrowth in humans[J].Nature,2000,403:383-384.
[26]Chen MS,Huber AB,Van der Haar ME et al.Nogo-A is a myelin-associated Neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-a[J].Nature,2000,403:434-439.
[27]Brittis PA,Flanagan JG.Nogo domains and a Nogo receptor implications for axon regeneration[J].Neuron,2001,30:11-14.
[28]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[J].Exp Neuro,2001,169:319-328.
[29]Huber AB,Weinman,Bramle C et al.Patterns of Nogo mRNA and protein expression in the developing and adult ratand after CNS lesions[J].Neurosci,2002,22:3553-3567.
[30]闵嵘,赵志炜,姬志娟,等.Nogo与Nogo受体在正常小鼠大脑的分布.神经解剖学杂志,2005,21(2):195-198.
[31]Wang X,Chun SJ,Treloar H,et al.Localization of Nogo-A and Nogo-66receptor proteins at sites of axon myelin and synaptic contact[J].Neurosci,2002,22(13):5505-5515.
[32]Fournier AE,Could GC,Liu BP,et al.Truncated soluble Nogo receptor binds Nogo-66 and blocks inhibition of axon growth by myelin[J].Neurosci,2002,22(20):8876-8883.
[33]Schwab ME.Myelin-associated inhibitors of neurite growth and regeneration in the CNS[J].Trends Neurosci,1990,13(11):452-456.
[34]Schwab ME,Caroni P.Oligodendrocytes and CNS myelin are nonperm issive substrates for neurite growth and fibroblast spreading invitro[J].J Neurosci,1988,8:2381-2393.
[35]Fournier AE,GrandPre T,Strittmatter SM.Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration[J].Nature,2001,409:341-346.
[36]Hunt D,Mason MR,Campbell G,et al.Nogo receptor mRNA expression in intact and regenerating CNS neurons[J].MolCell Neurosci,2002,20(4):537-542.
[37]Wang KC, Kim JA,Sivasankaran R, et al.P75 interacts with the Nogo receptor as a coreceptor for Nogo, MAG and OMgp[J]. Nature, 2002, 420(6911): 74-78.
[38] Niederost B, Oertle T, Fritsche J, et al.Nogo-A and myelin-associated glycoprotein mediate neurite growth inhibition by antagonistic regulation of RhoA and Racl[J].J Neurosci, 2002, 22(23):368-376.
[39]Yamashita T,Higuchi H, Tohyama M. The p75 receptor transduces the signal from myelin-associated glycoprotein to Rho[J]. JCell Biol, 2002, 157: 565-570.
[40]Skaper SD,Moore SE, WalshM F. Cell signaling cascades regulating neuronal growth promoting and inhibitory cues[J]. Prog Neurobiol, 2001, 65:593-608.
[41]Yuan XB, Jin M, Xu X, et. al. Signalling and crosstalk of Rho GTPases in mediating axon guidance [J]. Nat Cell Biol, 2003, 5: 38-45.
[42]Fournier AE , Takizawa BT , Strittmatter SM. Rho kinase inhibition enhances axonal egeneration in the injured CNS[J]. Neurosci , 2003, 23 : 1416-1423.
[43] Dergham P, Ellezam B, Essagian C et al. Rho signaling pathway targeted to promote spinal cord repair[J].J Neurosci, 2002, 22:6570-6577.
[44] He Z, Kop rivica V. The Nogo signaling pathway for regeneration block [J].Ann Rev Neurosci,2004,27:341-368.
[45] Winton MJ, Dubreuil CI, Lasko D,et al. Characterization of new cell permeable C3-like proteins that inactivate Rho and stimulate neurite outgrowth on inhibitory substrates[J]. J Biol Chem, 2002, 277:32820-32829.
[46]Neumann S, Bradke F, Tessier-LavigneM, et al. Regeneration of sensory axonswithin the injured sp inal cord induced by intraganglionic cAMP elevation[J]. Neuron, 2002, 34:885-893.
[47]Dergham P, Ellezam B, Essagian C, et al. Rho signaling pathway targeted to promote spinal cord repair[J].J Neurosci, 2002, 22:6570-6577.
[48]Grandpre T,Li S,Strittmatter SM.Nogo-66 receptor antagonist peptide promotes axonal regeneration[J].Nature,2002,417:547-551.
[49]王华,姚裕家.缺氧缺血新生鼠损伤脑组织Nogo-A,Ng-R mRNA的表达[J].郑州大学学报(医学版),2005,40(5):305-307.
[50]Teng FY,Tang BL.Why do Nogo/Nogo-66 receptor gene knockouts result in inferior regeneration compared to treatment with neutralizing agents[J].Neurochem,2005,9(5):865-874.
[51]张涛,袁文,刘百峰,等.SiRNA干扰大鼠神经元Nogo受体mRNA表达的时程研究[J].中国脊柱脊髓杂志,2005,15(6):588-591.
[52]陈志刚,朱诚,张光霁等 神经节苷脂GM-1对急性颅脑损伤的早期保护作用[J]中华神经外科杂志,1998,1(14):213-215.
[2]周伟.缺氧缺血性脑损伤的分子生物学进展[J].国外医学儿科学分册,1998,25(1):32-35.
[3]李炜如.兴奋性氨基酸及钙离子内流在缺氧缺血性脑损伤中的作用[J].中国实用儿科学杂志,1997,12(4):245-246.
[4]王冰,周文强,周从乐.新生鼠缺氧缺血后脑组织超氧化物歧化酶活性变化[J].新生儿科杂志.2002,17(1):12-13.
[5]Fellman V,Riavio KO.Reperfusion injury as the ischemic cerebral cell death[J].Prog Cardiovasc Dis,1999,42(3):185.
[6]Goddard DR,Berry M,Kirvell SL,et al.Fibroblast growth factor-2induces astroglial and microglial reaction in vivo[J].J Anat,2002,200(3):57-67.
[7]Zaremba J.Contribution of tumor necrosis factor alpha to the pathogenesis of stoke[J].Folia Morphol(warsz),2000,59(3):137-143.
[8]欧阳小琳.新生儿缺氧缺血性脑病血液酸碱失调分析及处理[J].中国实用儿科杂志,1999,14(5):297-298.
[9]薛辛东,杜立中主编.儿科学,北京:人民卫生出版社,2005:121-122.
[10]张少丹,钱培德,张淑琴.纳洛酮治疗新生儿缺氧缺血性脑损伤的临床研究[J].中国实用儿科杂志,1996,11(4):207-208.
[11]韩玉昆.脑细胞代谢激活剂在新生儿缺氧缺血性脑病治疗中的评价[J].中国实用儿科杂志,1998,13(2):80-82.
[12]张伟,吴婉芳,李松等.高压氧治疗新生儿缺氧缺血性性脑病疗效及毒副作用观察[J].中华儿科杂志,1997,12(1):13.
[13]陈志刚,朱诚,张光霁等神经节苷脂GM-1对急性颅脑损伤的早期保护作用[J].中华神经外科杂志,1998,1(14):213-215.
[14]Rowntree S,Kolb B.Blockade of basic fibroblast growth factor retards recovery from motor cortex injury in rats[J].EurJ Neutosci,1997,9:2432-2441.
[15]David S,Aguayo AJ.Axonal elongation into peripheral nervous system bridges after central nervous system injury in adult rats[J].Science,1981,214:931-933.
[16]Yiu G,He Z.Signaling mechanisms of the melin inhibitors of axon regeneratoion[J].Curr Opin Neurobiol,2003,13:1-7.
[17]Liu BP,Fournier A,Grandpre T,et al.Myelin-associalted glycoprotein asa functional ligand for the Nogo-66 receptor[J].Science,2002,297(5584):1190-1193.
[18]Caroni P,Schwab ME.Two membrane Protein factions from rat central myelin with inhibitory properties for neurite grouth and fibroblast spreading[J].Cell Biol,1988,106(4):1281-1284.
[19]Huber AB,Schwab ME.Nogo-A,a potent inhibitor of neurite outgrouth and regeneration[J].Biol Chem,2000,381:407-419.
[20]Perry AB,Flanagan JG.Uogo domains and a Nogo receptor implications for axonregeneration[J].Neuron,2001,30:11-14.
[21]陈益存,陆东东,张锡然.RTNs家族研究进展[J].细胞生物学杂志,2005,27:53-56.
[22]Grandpre T,Wakamura·F,Vartaninan T et al.Identification of the Nogo inhibitor of axon regeneration as a reticulon protein[J].Nature,2000,403:439-443.
[23]Oertle T,Van der Haar ME,Bandtlow CE et al.Nogo-A inhibits neurite outgrowth and cell spreading with three discrete regions[J].Neurosci,2003,23(13):5393-5406.
[24]Jin WL,Lin YY,Lin HL et al.Intraneuronal location of Nogo-A in the rat[J].Comp Neurol,2003,458:1-10.
[25]Prinjha R,Morrow SE,Vinson M et al.Inhibitor of neurite outgrowth in humans[J].Nature,2000,403:383-384.
[26]Chen MS,Huber AB,Van der Haar ME et al.Nogo-A is a myelin-associated Neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-a[J].Nature,2000,403:434-439.
[27]Brittis PA,Flanagan JG.Nogo domains and a Nogo receptor implications for axon regeneration[J].Neuron,2001,30:11-14.
[28]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[J].Exp Neuro,2001,169:319-328.
[29]Huber AB,Weinman,Bramle C et al.Patterns of Nogo mRNA and protein expression in the developing and adult ratand after CNS lesions[J].Neurosci,2002,22:3553-3567.
[30]闵嵘,赵志炜,姬志娟,等.Nogo与Nogo受体在正常小鼠大脑的分布.神经解剖学杂志,2005,21(2):195-198.
[31]Wang X,Chun SJ,Treloar H,et al.Localization of Nogo-A and Nogo-66receptor proteins at sites of axon myelin and synaptic contact[J].Neurosci,2002,22(13):5505-5515.
[32]Fournier AE,Could GC,Liu BP,et al.Truncated soluble Nogo receptor binds Nogo-66 and blocks inhibition of axon growth by myelin[J].Neurosci,2002,22(20):8876-8883.
[33]Schwab ME.Myelin-associated inhibitors of neurite growth and regeneration in the CNS[J].Trends Neurosci,1990,13(11):452-456.
[34]Schwab ME,Caroni P.Oligodendrocytes and CNS myelin are nonperm issive substrates for neurite growth and fibroblast spreading invitro[J].J Neurosci,1988,8:2381-2393.
[35]Fournier AE,GrandPre T,Strittmatter SM.Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration[J].Nature,2001,409:341-346.
[36]Hunt D,Mason MR,Campbell G,et al.Nogo receptor mRNA expression in intact and regenerating CNS neurons[J].MolCell Neurosci,2002,20(4):537-542.
[37]Wang KC, Kim JA,Sivasankaran R, et al.P75 interacts with the Nogo receptor as a coreceptor for Nogo, MAG and OMgp[J]. Nature, 2002, 420(6911): 74-78.
[38] Niederost B, Oertle T, Fritsche J, et al.Nogo-A and myelin-associated glycoprotein mediate neurite growth inhibition by antagonistic regulation of RhoA and Racl[J].J Neurosci, 2002, 22(23):368-376.
[39]Yamashita T,Higuchi H, Tohyama M. The p75 receptor transduces the signal from myelin-associated glycoprotein to Rho[J]. JCell Biol, 2002, 157: 565-570.
[40]Skaper SD,Moore SE, WalshM F. Cell signaling cascades regulating neuronal growth promoting and inhibitory cues[J]. Prog Neurobiol, 2001, 65:593-608.
[41]Yuan XB, Jin M, Xu X, et. al. Signalling and crosstalk of Rho GTPases in mediating axon guidance [J]. Nat Cell Biol, 2003, 5: 38-45.
[42]Fournier AE , Takizawa BT , Strittmatter SM. Rho kinase inhibition enhances axonal egeneration in the injured CNS[J]. Neurosci , 2003, 23 : 1416-1423.
[43] Dergham P, Ellezam B, Essagian C et al. Rho signaling pathway targeted to promote spinal cord repair[J].J Neurosci, 2002, 22:6570-6577.
[44] He Z, Kop rivica V. The Nogo signaling pathway for regeneration block [J].Ann Rev Neurosci,2004,27:341-368.
[45] Winton MJ, Dubreuil CI, Lasko D,et al. Characterization of new cell permeable C3-like proteins that inactivate Rho and stimulate neurite outgrowth on inhibitory substrates[J]. J Biol Chem, 2002, 277:32820-32829.
[46]Neumann S, Bradke F, Tessier-LavigneM, et al. Regeneration of sensory axonswithin the injured sp inal cord induced by intraganglionic cAMP elevation[J]. Neuron, 2002, 34:885-893.
[47]Dergham P, Ellezam B, Essagian C, et al. Rho signaling pathway targeted to promote spinal cord repair[J].J Neurosci, 2002, 22:6570-6577.
[48]Grandpre T,Li S,Strittmatter SM.Nogo-66 receptor antagonist peptide promotes axonal regeneration[J].Nature,2002,417:547-551.
[49]王华,姚裕家.缺氧缺血新生鼠损伤脑组织Nogo-A,Ng-R mRNA的表达[J].郑州大学学报(医学版),2005,40(5):305-307.
[50]Teng FY,Tang BL.Why do Nogo/Nogo-66 receptor gene knockouts result in inferior regeneration compared to treatment with neutralizing agents[J].Neurochem,2005,9(5):865-874.
[51]张涛,袁文,刘百峰,等.SiRNA干扰大鼠神经元Nogo受体mRNA表达的时程研究[J].中国脊柱脊髓杂志,2005,15(6):588-591.
[52]陈志刚,朱诚,张光霁等 神经节苷脂GM-1对急性颅脑损伤的早期保护作用[J]中华神经外科杂志,1998,1(14):213-215.