摘要
目的:通过针刺对家兔坐骨神经损伤(SNI)后Nogo-A蛋白和髓磷脂相关糖蛋白(MAG)影响的实验研究,观察针刺对周围神经损伤的治疗作用,进一步探讨其作用机理,为临床治疗提供新的理论依据。
方法:选用48只家兔随机分为3组(每组16只):空白对照组、模型对照组、针刺治疗组。模型对照组、针刺治疗组给予手术钳夹造成家兔坐骨神经SunderlandⅡ~Ⅲ度损伤模型。针刺治疗组在造模后第2天开始针刺“环跳”、“足三里”穴。采用平补平泻捻转手法,每次捻转30秒,间隔10分钟捻转一次,20分钟后起针。每日1次,7天为1疗程,休息1天;模型对照组不做针刺治疗,但与针刺组同样进行动物抓取;空白对照组不做任何处理。分别于第1、2疗程治疗结束后当天观察各组家兔右后肢的功能状况,检测三组家兔相应脊髓节段中Nogo-A蛋白和MAG含量,并对结果进行比较。
结果:在基本功能改善方面,与模型对照组比较,针刺治疗组家兔右后腿功能明显改善。脊髓前角Nogo-A蛋白变化:1个疗程后,针刺治疗组明显高于空白对照组,组间具有显著性差异(P<0.05),而低于模型对照组,组间具有显著性差异(P<0.05);2个疗程后,针刺治疗组明显低于模型对照组,组间具有显著差异(P<0.05),与空白对照组没有显著性差异(P>0.05)。脊髓前角MAG变化:1个疗程后,针刺治疗组与模型对照组没有显著性差异(P>0.05),而明显高于空白组(P<0.05);2个疗程后,针刺治疗组低于模型对照组,组间具有显著性差异(P<0.05),而高于空白对照组(P<0.05)。
结论:通过本实验研究发现,针刺能改善坐骨神经急性损伤家兔的一般状况,针刺组家兔脊髓中Nogo-A蛋白及MAG含量明显减少,说明针刺能抑制脊髓中抑制性生长因子的释放,有助于触突的生长,促进周围神经损伤再生修复。
Objective: To observe the therapeutic effect of treating peripheral nerve injury with acupuncture and provide new theoretical basis for clinic by experimental research of Nogo-A protein and myelin-associated glycoprotein(MAG) after sciatic nerve injury(SNI) in rabbits.
Methods: 48 healthy adult rabbits were randomly divided into 3 groups: blank control group, model control group and acupuncture treatment group. Except blank control group, the right sciatic nerve injury(SNI) model of SunderlandⅡ~Ⅲdegree were made in other two groups by operation .One day after operation, we punctured Huantiao(GB 30) and Zusanli(ST 36) of the affected side in EA group, and applied even reinforcing-reducing by twirling-rotating manipulation, twisted it for 30 seconds every time and moved it one time per 10 minutes, retained the needle for 20 minutes, once daily, seven treatments made a course and there was one day break between two courses. The model control group was grasped every day but with no treatment. We observed the function of posterior right paw of all animals and examined the content of Nogo-A protein and MAG in the corresponding spinal cord segment.
Results:(1) After finishing the treatment, Function of injured limbs were obviously improved acupuncture treatment group. (2) After a therapeutic course, the content of Nogo-A protein in acupuncture treatment group were higher than that of blank control group while lower than model control group, there was significant difference (P<0.05); two courses later, the content of Nogo-A protein in acupuncture treatment group was significantly lower than that of model control group(P<0.05) and there was no significant difference with blank control group (P>0.05). (3)The content of MAG in anterior horn of spinal cord: a therapeutic course later there was no significant difference between acupuncture treatment group and model control group(P > 0.05), while significantly higher than that of model control group(P<0.05); after treating for 14 days, the content of MAG in acupuncture treatment group was lower than that of that in model control group, there was a significant difference between groups (P<0.05) ,while remarkably higher than that of blank control group(P<0.05).
Conclusion: Our research showed that acupuncture treatment could improve the functional recovery of acute SNI in rabbits, restrain the release of inhibitory growth factor in spinal cord, promote the growth of axon and the regeneration of affected nerve.
引文
[1]肖桂荣,郝华,赵秋玲等.电针并功能训练治疗上肢周围神经不完全损伤.中国针灸,2007;(27):329-332
[2]王东雁.电针治疗周围神经损伤30例.中国针灸,1997;(12):726
[3]袁广宇,苗小军,王培霞.电针加康复治疗腓总神经损伤86例疗效观察.中国实用神经疾病杂志,2008;(11):35-36
[4]张志良.电针治疗四肢神经损伤性瘫痪100例.针灸临床杂志,1995;(1):33
[5]田成举.针刺治疗臂丛神经损伤97例.中国针灸,1994;(1):10
[6]解霖源.针灸治疗腓总神经损伤40例报告.中医杂志,1982;(9):43-44
[7]赵健.针灸按摩治疗臀上皮神经损伤120例疗效观察.中国针灸,1989(;3):18-19
[8]王丽丽.针刺治疗新生儿丛神经麻100例.中国针灸,1989;(5):4
[9]张慰民.针刺治疗臀上皮N损伤64例.针灸学报,1990;(3):23
[10]魏彩莲.针刺治疗腓神经麻痹50例.中国针灸,1996;(12):38
[11]刘远珍,王素娥.阳明经穴位注射治疗腓总神经损伤的疗效观察.中国现代医学杂志,2008;18(19):2891-2892
[12]郭志权,严锡峰.穴位注射治疗儿童腓总神经损伤16例疗效观察.海军医学杂志,2008;29(2):130-131
[13]梁中,刘敏玲,王鸿章.电针配合穴位注射弥可保治疗新生儿臂丛神经麻痹疗效观察,针灸骨伤,2007;29(7):53
[14]任善玲.穴位注射治疗臂丛神经麻痹.山东中医杂志,2006;25(1)9
[15]潘玉印.穴位注射治疗臂丛神经损伤37例,中国针灸,1996;16(3):24
[16]钟泽鑫,吴清良.穴位注射治疗坐骨神经痛200例.福建中医药,1999;30(3):41-42
[17]陈荣.湿针治疗腓总神经损伤13例.四川中医.2000;18(7):53
[18]张连生.温针治疗坐骨神经痛518例.上海针灸杂志,1998;17(3):33
[19]肖凌勇,孙申田,李征.头针电针结合治疗腓总神经麻痹的临床观察.针灸临床杂志,2008;24(2):19-20
[20]孙怀玲,姚广珍,李兰香.头针治疗坐骨神经痛100例.针灸临床杂志,1999;15(9):51-52
[21]吴小明,董祖木.刺血拔罐治疗坐骨神经痛66例.陕西中医,1999;20(6):286
[22]杜艳娟.电针配合隔姜灸治疗腓总神经麻痹24例.中国针灸杂志,2008;28(4):296
[23]牛凤景,韩宝生,郭同安.隔姜灸治疗坐骨神经痛18例.上海针灸杂志,1998;17(3):34
[24]周伟平,祝维华.穴位埋线治疗腰椎增生并坐骨神经痛156例。江西中医药,1999;30(4):46
[25]Bisby MA,Bulger VT.Reversal of axonal transport at a nerve crush.J Neurochem,1997;29:313
[26]Black MM,Lasek RJ.slow components of axonal transport two cytoskeletal networks.J Cell Biol,1980;85:616
[27]胡玮,周梁.睫状神经营养因子与神经再行.中华耳鼻喉科杂志,1998;33(6):377-379
[28]李陶,陈意生.睫状神经营养因子雪旺氏细胞与周围神经再生.国外医学临床生物化学与检验分册,2001;22(1):25-27
[29]李陶,陈意生.睫状神经营养因子雪旺氏细胞与周围神经再生,国外医学临床生物化学与检验分册,2001;22(1):25-27
[30]Woolf CJ,Bloechlinger S.It takes mor than two to Nogo Science,2002 ;297:1132-1134
[31]Pot C,Simonen M,Weinmann O et al Nogo-A expressed in Schwann cell impairs axonal regeneration after peripheral nerve injury.J Cell Biol,2002;159:29-35
[32]Ng CE, Tang BL, Tang BL. Nogos and Nogo-66 recep tor: factors inhibiting CNS neuron regeneration Neurosci Res, 2002;67: 559-565
[33]DeBellard M E,Tang S,Mukhopadhyay G,et al M yelin-associated glycoprotein inhibits axonal regeneration from a variety of neurons via interaction with a sialoglycoprotein[J].Mol Cell Neurosci,1996;7(2):89-101
[34]Mukhopadhyay G,Doherty P,Walsh FS,et al A novel role form yelin-associated glycoprote in as an inhibitor of axonal regeneration[J].Neuron,1994;13(3):757-767
[35]Martini R.Expression and functional roles of neural cellsur-face molecules andextracellu larmatrix com ponents suring sevelopment and regeneration of peripheral nerves[J].J Neurocytol,1994;23(1):1-28
[36]Filbin MT.The muddle with MAG[J].Mol Cell Neurosci,1996,8(2-3):84-92
[37]Mukhopadhyay G,Doherty P,Walsh FS,et al A novel role form yelin-associated glycoprote in as an inhibitor of axonal regeneration[J].Neuron,1994;13(3):757-767
[38]McKerracher L,David S,Jackson DL,et al Identification of mylinassociated glycoprotein as a majorm yelin-derived inhibitor of neurite growth[J].Neuron,1994;13(4):805-811
[39]陈春林,叶剑.髓磷脂相关性轴突生长抑制因子与中枢神经再生,创伤外科杂志2005;7(5):391-393
[40]Mingorance A,Fontana X,Soriano E,et al Overexpression of m elin-associated glycoprotein after axotomy of the perforant pathway[J].Mol Cell Neurosci,2005;29(3):471-483
[41]屈红艳,王瑞辉,刘飞虎.电针对家兔坐骨神经损伤后IL-1β的影响.陕西中医,2007;28(4):506
[42]李忠仁主编.实验针灸学,北京:中国中医药出版社,2003,第1版:415
[43]牙祖蒙,王建华,周渝,等.穴位针刺对面神经再生影响的动物模型制作[J].实验动物科学与管理,1999;16(1):15
[44]杨金蓉,宋军,田丰伟等.管灸对面神经损伤模型家兔面神经核NGFmRNA表达的影响.中国中医基础医学杂志,2008;14(9):703
[45]孙忠人,梅晨健,宋春华,等.针刺对大鼠坐骨神经损伤模型组织形态学改变影响的实验研究,中国中医药科技,1998;5(1):1
[46]李岩,李平,周震,等.火针对坐骨神经损伤模型大鼠COX-2、Ll-1β、BDNF表达的干预研究.针灸临床杂志,2007;23(12):36
[47]陈敏,陈利民,项红兵,等.曲马多对慢性压迫性坐骨神经损伤模型大鼠脊髓突触重塑的影响.中国临床康复,2006;10(40):63
[48]聂勇,蔡国平.透明质酸酶对慢性压迫性神经损伤模型中神经源性疼痛的作用研究.针刺研究,2005;30(2):88
[49]孙忠人,金淑英,孙远征,等.针刺对周围神经损伤再生修复的机理研究.中国中医药科技,1996;3(2):10
[50]邵水金,单宝枝,严振国,等.电针促进大鼠坐骨神经再生的实验研究.现代康复,2000;4(8):184
[51]吴克坚,殷代昌,陈家臻.周围神经损伤的实验与临床观察,临床军医杂志,2001;29(1):58
[52]王相利,杨琳,李振华等.“补阳还五汤”对周围神经再生影响的实验研究.山东医科大学学报,1999;37(1):41
[53]徐欣,马跃,李茜等.膜引导组织再生术与硅胶管桥接法修复兔面神经缺损.上海口腔医学,2000;9(3):156
[54]李书林,蒋晓江,陈曼娥.大鼠坐骨神经损伤后髓鞘碱性蛋白与神经功能的变化及神经生长因子的影响.中国临床康复,2002;6(6):798
[55]NA ISHI Y,KASHIHARA Y,SAKAGUCHIM,et al Cooperative regulation of calcitonin gene-related peptide levels in rat sensory neurons via their central and peripheral processes[J].J Neurosci,1992;12(2):518-524
[56]蔡文琴,阮怀珍,黎海蔕.医用神经生物学基础[M].西南师范大学出版社,2001;41:416-420
[57]邱文兴,段淑香,侯林萍,等.环跳穴与坐骨神经的解剖学研究.针灸临床杂志,1999;15(4):57
[58]杨新东,楼新法,蒋松鹤.足三里穴的断面解剖学研究.温州医学院学报,2000;30(3):224
[59]邵金水,等.周围神经损伤的针灸疗法.现代康复,2000;11(4):1616-1617
[60]胡艳影.针灸治疗周围神经损伤近况.针灸临床杂志,1998;14(10):46-50
[61]周洁,等.齐刺法及其临床应用.江西中医药,2000;31(5):36-38
[62]李澎涛,等.电针、神经营养因子影响周围神经再生形态与功能恢复的比较分析.中国针灸,1999;9:556-559
[63]孙忠人,等.针刺对周围神经损伤再生修复的机理研究—针刺对坐骨神经损伤诱发电位的影响.中国中医药科技,1996;3(2):10
[64]隋莲方.电针促进小鼠坐骨神经损伤后修复.上海针灸杂志,1994;13(4):174-175
[65]邵金水,等.中药对大鼠坐骨神经损伤后SFI及肌肉影响的观察.中国针灸1998;(2):249-251
[66]Josephson A, Trifunovski A,Widmer HR,et al.Rogo-receptor gene activity:cellular localization and developmental regulation of mRNA in mice and humans [J].J Comp Neurol,2002;453(3):292
[67]Hunt D,Mason M,Campebell G,et al.Nogo receptor mRNAexpression in intact and regenerating CNS neurons [J].Mol Cell Neurosxi,2002;20(1153):537
[68]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
[69]Caroni P.Schwab ME.Two membrane protein factions from rat central myelin with inhibitory properties for neurite growth and fibroblast spreading.J Cell Biol,1988;106(4):1
[70]Mckerracher L,David S,Jackson DL,et al.Identification of myelin-associated glycoprotein as a major myelin-derivd inhibitor of neurite growth.Neuron,1994;13(4):805-1
[71]Li M,Shibata A,Li C,et al.Myelin-associated glycoprotein inhibits neurite/axon growth and causes growth cone collapse.J Neurosci Res,1996;46(4):404-14
[72]Prinjha R,Moore SE,Vinson M,et al.Inhibitor of neurite outgrowth in humans.Nature,2002;403:383-4
[73]Clifford J W,Stefan B.It takes more than two to Nogo.Science,2002;16:1 132-4
[74]David S,Aguayo AJ.Axonal elongation into peripheral nervous system bridges after central nervous system injury in adult rats.Science.1981,214:931-33
[75]陈春林,叶剑.髓磷脂相关性轴突生长抑制因子与中枢神经再生,创伤外科杂志2005;7(5):391-393
[76]Quarles R H.M yelin-associated glycoprotein (MAG):past present and beyond [J].J N eurochem,2007,100(6):1431-1448
[77]Quarles R H.M yelin-associated glycoprotein (MAG):past present and beyond [J].J N eurochem,2007,100(6):1431-1448
[78]Filbin M T.The muddle with MAG[J].Mol Cell N eurosci,1996,8(2-3):84-92
[79]庞超见,贾桦,李振华.髓磷脂相关糖蛋白对周围神经影响的研究进展.解剖学研究.2008;30(4):306-308
[80]DeBellard M E,Tang S,Mukhopadhyay G,et al Myelinassociated glycoproteininhibits axonal regeneration from a variety of neurons via interaction with a sialoglycoprotein[J].Mol Cell Neurosci,1996;7(2):89-101
[81]DeBellard M E,Tang S,Mukhopadhyay G,et al Myelinassociated glycoprotein inhibits axonal regeneration from a variety of neurons via interaction with a sialoglycoprotein[J].Mol Cell Neurosci,1996;7(2):89-101
[82]MartiniR.Expression and functional roles of neural cell surface molecules and extracellular matrix components during development and regeneration of peripheral nerves [J].J Neurocytol,1994;23(1):1-28
[83]Adele G,Woolley,Katen J,et al.Developmental loss of NT-3 in vivo results in reduced levels of myelinspecific proteins,a reduced extent of myelination and increased apoptosis of Schwann cells.Glia,2008;56:306-317