摘要
背景
急性脊髓损伤(spinal cord injury，SCI)是指脊髓遭受到外力打击后导致完全性或不完全性脊髓运动、感觉、括约肌功能障碍等，是脊柱外科领域中一种常见且破坏性极大的疾病。随着脊髓损伤基础及临床研究的不断深入，脊髓损伤的治疗也得到不断发展。近年，不同学者采用不同方法治疗脊髓损伤病人，并取得一定的效果，也有报道一些药物有助于脊髓损伤后功能恢复。然而所有结果仍不尽人意。为寻求一种新的治疗脊髓损伤的方法，根据脊髓损伤后病理改变，最大限度减少这种病理改变将是一种有效治疗方法。全身性应用药物通过血循环途径达到脊髓损伤部位，可能会有效的打断脊髓损伤后病理进程从而达到治疗脊髓损伤、保护或者恢复脊髓功能的目的。为此，进一步研究一种较好的用药品种及用药程式将是一种非常有前景的方法。
目的
1．探讨应用药物治疗脊髓损伤行为学的改变；
2．探讨应用药物治疗脊髓损伤病理学的改变；
3．探讨应用药物治疗脊髓损伤神经细丝及胶质纤维酸性蛋白表达的影响；
4．探讨应用药物治疗脊髓损伤对细胞凋亡的影响。
5．通过结果探讨不同药物对大鼠急性脊髓损伤的治疗作用，为临床上应用药物治疗脊髓损伤提供理论指导。
方法
实验中将216只大鼠分为对照组及6种不同药物组。采用腹腔内药物注射进行麻醉后，利用改良Allell's打击法建立大鼠急性脊髓损伤模型。手术后通过尾静脉给予药物。在术后8h、24h、4天、7天、14天分别进行以下指标检测：1、采用胥少汀对脊髓损伤的四肢行走动物提出的后肢6级行为学评分障碍率及
BACKGROUND
Acute spinal cord injury(SCI) is defined as an insult to the spinal cord that partially or completely interrupts the three main functions of the cord-motor, sensory, and reflex activities. It is mostly caused by motor vehicle accidents, falls, sports injuries and some other wounds. During years of unremitting efforts by scientists of many countries, with basic and clinical research of the spinal cord injury, treatment to the spinal cord injury has been developed much. However, it is not satisfactory. Considering the process of SCI, to decrase or to block the process may be an effective method. Though some drugs showed much effect in proctecting and improving the spinal function after injury, searching for an better means to improve its function is still an important subject to be solved.
OBJECTIVE
To study the effect of different drugs to the behavior and neurological changes , pathological and ultrastructural changes , the expression of neurofilament (NF) and glial fibrillary acidic protein (GFAP) ,and nerve cell apoptosis of spinal cord in rats after spinal cord injury. Then to find an effective method to cure SCI.
METHODS
Two hundreds and sixteen healthy adult Sprague-Dawley rats were randomly divided into seven groups:blank control group, control group, experimental group. Build acute spinal cord injury model in rats with modified Allen's method after rats were anesthesed with drug administration through abdomin. Different drugs were given to cure spinal cord injury through tail vein each day. The follow index were observed at 8h、24h、96h、7day、14day:(1) to evaluate the function of spinal cord by
引文
1．胥少汀，等．脊髓损伤基础与临床．人民卫生出版社，2002
2. Rivilin AS, Tator CH. Objective clinical assessment of motor function after experiment spinal cord injury in the rat. J Neurosurg, 1997,47:577-581.
3. Fukaya C, Katayama Y, Kasai M, et al. Evaluation of time-dependent spread of tissue damage in experimental spinal cord injury by killed-end evoked potential: effect of high-dose methylprednisolone. J Neurosurg. 2003 Jan;98(1 Suppl):56-62
4. Lankorst AJ, ter Laak MP, et al. Combined treatment with alpha MSH and methyprednisolone fails to improve functional recovery after spinal cord injury in the rat. [J] Brain Res2000;859(2):334-40
5. Liu D, Li L, et al. Prostaglandin release by spinal cord injury mediates production of hydroxyl radical, malondiadehyde and cell death: asite of neuroprective action of methylpredinsolone [J]. Neurotraum, 2001;77(4):1036-47
6. Hsu CY. Dimitrijevic MR. Methylperdnisolone and spinal cord injury: the possible mechanism of action. J Neurotrauma. 1990 Fall; 7(3): 115-9.
7. Anderson DK, Means ED. Iron-induced lipid peroxidation in spinal cord: protection with mannitol and methylprednisolone. J Free Radic Biol Med. 1985;1(1):59-64.
8. Hall ED, Yonkers PA, Tayler BM. Lack of effect of postinjury treatment with MP or tirilared mesylate on the increase in eicosanoid levels in the aculely injured cat spinal cord. J Neuro Trauma, 1995;12(3):245.
9. Garcia,-M;Forster,-V;Hicks,-Detal.In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcineretina in vitro. Invest-Ophthalmol-Vis-Sci. 2003;44(10):4532～4541
10. Shimode,-H;Ueki,-A;Morita,-Y. Nerve growth factor attenuate ship pocampal choliner gicdeficits and operant learning impairment in rats with entorhinal cortex lesions. Behav-Pharmacol. 2003; 14(3): 179～190
11．林剑梅，曹向光，左平英。β—七叶皂甙钠的药理作用及国内应用概况。中国药师，2003，6(7)：445—448。
12．崔丽，郑惠民，倪灿荣，等．七叶皂甙钠对大鼠脑缺血损伤中神经细胞凋亡的影响．药学服务与研究．2002(01)
13. Lu HS Pharmacological role and clinical application of β-Sodium Aescinate. Yiyao Daobao,1994,13(2):71-2.
1．胥少汀，郭世绂．脊髓损伤基础与临床．人民卫生出版社，2002．
2．方健，陈文笔，严家春等脊髓损伤大鼠脊髓组织的病理形态学观察。中国矫形外科杂志，2002，10(10)：996—970。
3．郑清波，施杞，姜立本 大鼠脊髓受压后的病理学变化。中国骨伤，1999，12(5)：22—26。
4. Fukaya C, Katayama Y, Kasai M, et al. Evaluation of time-dependent spread of tissue damage in experimental spinal cord injury by killed-end evoked potential: effect of high-dose methylprednisolone. J Neurosurg. 2003 Jan;98(1 Suppl):56-62
5. Liu D, Li L, et al. Prostaglandin release by spinal cord injury mediates production of hydroxyl radical, malondiadehyde and cell death: asite of neuroprective action of methylpredinsolone [J]. Neurotraum, 2001;77(4):1036-47
6. Hsu CY. Dimitrijevic MR. Methylperdnisolone and spinal cord injury: the possible mechanism of action. J Neurotrauma. 1990 Fall; 7(3): 115-9.
7. Anderson DK, Means ED. Iron-induced lipid peroxidation in spinal cord: protection with mannitol and methylprednisolone. J Free Radic Biol Med. 1985;1(1):59-64.
8. Hall ED, Yonkers PA, Tayler BM. Lack of effect of postinjury treatment with MP or tirilared mesylate on the increase in eicosanoid levels in the aculely injured cat spinal cord. J Neuro Trauma, 1995;12(3):245.
9. Garcia,-M;Forster,-V;Hicks,-Detal.In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcineretina in vitro. Invest- Ophthalmol-Vis-Sci. 2003;44(10):4532～4541.
10. Shimode,-H;Ueki,-A;Morita,-Y. Nerve growth factor attenuate ship pocampal choliner gicdeficits and operant learning impairment in rats with entorhinal cortex lesions. Behav-Pharmacol. 2003;14(3):179～190.
11. Garcia,-M; Forster, V;Hicks,-Detal. In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcine retina in vitro. Invest- Ophthalmol- Vis-Sci. 2003;44(10):4532～4541.
12．王金光，郑启新，郭晓东等 应用β七叶皂甙钠治疗大鼠脊髓损伤的实验研究。中国康复医学杂志，2005，20(1)：15—18。
13．王金光，郑启新，郭晓东等 重组人红细胞生成素与β七叶皂甙钠治疗大鼠脊 髓损伤的实验研究。中国脊柱脊髓杂志，2003，13(11)：674—78。
14．林剑梅，曹向光，左平英。β—七叶皂甙钠的药理作用及国内应用概况。中国药师，2003，6(7)：445—448。
15．崔丽，郑惠民，倪灿荣，等．七叶皂甙钠对大鼠脑缺血损伤中神经细胞凋亡的影响．药学服务与研究．2002(01)
16. Lu HS Pharmacological role and clinical application of β-Sodium Aescinate. Yiyao Daobao,1994,13(2):71-2.
17. Shimode,-H; Ueki,-A;Morita, -Y. Nerve growth factor attenuates hippocampalcho liner gicdeficits and operant learning impairment in rats with entorhinal cortex lesions. Behav-Pharmacol. 2003;14(3):179～190.
1. Uchida K, Baba H, Maezawa Y, et al. Progressive changes in neurofilament proteins and growth-associated protein-43 immunoteactivities at the site of cervical spinal cord compression in spinal hyperostotic mice[J]. Spine,2002,27(5):480.
2. Zhang ZX, Casey DM, Julien JP, et al. Normal dentritilic arborization in spinal motoneurones reguires neurofilament subunit.[J]Comp Neurol, 2002, 450(2)144.
3. Yabe J T, Wang F S, Chylinski T, et al. Selective accumulation of the high molecular weight neurofilament subunit within the distal region of growing axonal neuritis[J]. Cell Motil Cytoskeleton,2001,50(1):1.
4. Brook G A, Plate D, Franzen R, et al. Spontaneous longitudinally orientated axonal regeneration is associated with the Schwann cell framework within the lesion site following spinal cord compression injury of the rat[J]. J Neurosci Res,1998,53(1):51.
4．胡俊勇，李佛保，廖威明，等．大鼠脊髓损伤后神经丝蛋白及胶质纤维酸性蛋白表达的变化及意义．中山大学学报(医学科学版)；2003；24(2)：129．
5．杨青峰，龙在云，曾琳，等．大鼠脊髓缺血损伤后神经微丝及胶质纤维酸性蛋白的改变．中华创伤杂志，1998，10(14)：40-42．
7．张卫红，王道新．脊髓损伤后胶质纤维酸性蛋白的表达及其意义．南京医科大学学报；2002，22(1)：17
8. Bigini P, Bastone A, Mennini T. Glutamate transporters in the spinal cord of the wobbler mouse[J]. Neuroreport, 2001,12(9):1815
9. Cheng H, Wu J P, Tzeng S F. Neuroprotection of glial cell line-derived neurotrophic factor in damaged spinal cords following contusive injury[J]. J Neurosci Res, 2002,69(3):397.
10. Kawai H, Arata N, Nakayasu H. Three-dimensional distribution of astrocytes in zebrafish spinal cord[J]. Glia, 2001,36(3):406.
11. Tatagiba M, Brosamle C, Schwab M. Regeneration of injured axons in the adult mammalian central nervous system. Neurosurgery;1997,40: 541-547.
12. Hinkle D, Baldwin S, Scheff S, et al. GFAP and S100beta expression in the cortex and hippocampus in response to mild cortical contusion.J Neurotrauma. 1997 Oct;14(10):729-38.
13. Laping NJ, Nichols NR, Day JR, et al. Transcriptional control of glial fibrillary acidic protein and glutamine synthetase in vivo shows opposite responses to corticosterone in the hippocampus. Endocrinology. 1994 Nov;135(5):1928-33.
1. Ekshyyan O, Aw TY.Apoptosis in acute and chronic neurological disorders. Front Biosci. 2004, 01 ;(9):1567-76.
2. Kim DH, Vaccaro AR, Henderson FC,et al.Molecular biology of cervical myelopathy and spinal cord injury: role of oligodendrocyte apoptosis. Spine J. 2003,3(6):510-519.
3. Li GL, Brodin G, Farooque M, et al. Apoptosis and expression of Bax after compression trauma to rat spinal cord injury. J Neuropathol Exp neurol, 1996, 55(3): 281-289.
4. Wingrave JM,Sribnick EA,Wilford GG,et al. Relatively low levels of calpain expression in juvenile rat correlate with less neuronal apoptosis after spinal cord injury. Exp Neurol. 2004;187(2):529-32.
5. Yune TY, Kim S J, Lee SM,etal.Systemic administration of 17beta-estradiol reduces apoptotic cell death and improves functional recovery following traumatic spinal cord injury in rats. J Neurotrauma. 2004 ;21(3):293-306.
6. Seki T, Hida K, Tada M,et al.Role of the Bax gene after contusive spinal cord injury in mice. Neurosurgery,2003,53(1): 192-198.
7. Fukaya C, Katayama Y, Kasai M, et al. Evaluation of time-dependent spread of tissue damage in experimental spinal cord injury by killed-end evoked potential: effect of high-dose methylprednisolone. J Neurosurg. 2003 Jan;98(1 Suppl):56-62
8. Shimode,-H;Ueki,-A;Morita,-Y. Nerve growth factor attenuate ship pocampal choliner gicdeficits and operant learning impairment in rats with entorhinal cortex lesions. Behav-Pharmacol. 2003; 14(3): 179～190.
9. Garcia,-M; Forster, V;Hicks,-Detal. In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcine retina in vitro. Invest- Ophthalmol- Vis-Sci. 2003;44(10):4532～4541.
10．王金光，郑启新，郭晓东等 应用β七叶皂甙钠治疗大鼠脊髓损伤的实验研究。中国康复医学杂志，2005，20(1)：15—18。
11．王金光，郑启新，郭晓东等 重组人红细胞生成素与β七叶皂甙钠治疗大鼠脊髓损伤的实验研究。中国脊柱脊髓杂志，2003，13(11)：674—78。
12．崔丽，郑惠民，倪灿荣，等．七叶皂甙钠对大鼠脑缺血损伤中神经细胞凋亡的影响．药学服务与研究．2002(01)
1．胥少汀，等．脊髓损伤基础与临床．人民卫生出版社，2002
2．胥少汀．脊髓损伤治疗进展[J]．引进国外医药技术与设备，2000；6(1)：29-33．
3. Kiwerski JE. Application of dexamethasone in the treatment of acute spinal cord injury. Injury 1993;24:457-60.
4. Bracken MB, Shepard MJ, Holford TR, et al. Administration of methylprednisolone for 24 or 48 hours or tirilazad mesylate for 48 hours in the treatment of acute spinal cord injury, Results of the Third National Acute Spinal Cord Injury Randomized Controlled Trial. JAMA, 1997,277:1597
5. Fukaya C, Katayama Y, Kasai M, et al. Evaluation of time-dependent spread of tissue damage in experimental spinal cord injury by killed-end evoked potential: effect of high-dose methylprednisolone. J Neurosurg. 2003 Jan;98(1 Suppl):56-62
6. Lankorst AJ, ter Laak MP, et al. Combined treatment with alpha MSH and methyprednisolone fails to improve functional recovery after spinal cord injury in the rat. [J] Brain Res2000;859(2):334-40
7. Liu D, Li L, et al. Prostaglandin release by spinal cord injury mediates production of hydroxyl radical, malondiadehyde and cell death: asite of neuroprective action of methylpredinsolone [J]. Neurotraum, 2001;77(4):1036-47
8. Hsu CY. Dimitrijevic MR. Methylperdnisolone and spinal cord injury: the possible mechanism of action. J Neurotrauma. 1990 Fall; 7(3): 115-9.
9．林剑梅，曹向光，左平英。β—七叶皂甙钠的药理作用及国内应用概况。中国药师，2003，6(7)：445—448。
10．崔丽，郑惠民，倪灿荣，等．七叶皂甙钠对大鼠脑缺血损伤中神经细胞凋亡的影响．药学服务与研究．2002(01)
11. Lu HS Pharmacological role and clinical application of β-Sodium Aescinate. Yiyao Daobao,1994,13(2):71-2.
12. Katoh D, Ikata T, Katoh S, et al. Effect of dietary vitamin C on compression injury of the spinal cord in a rat mutant unable to synthesize ascorbic acid and its correlation with that of vitamin E. Spinal Cord. 1996 Apr;34(4):234-8.
13. Fujimoto T, Nakamura T, Ikeda T,et al. Effects of EPC-K1 on lipid peroxidation in experimental spinal cord injury. Spine. 2000 Jan;25(1):24-9.
14. Anderson DK, Means ED. Iron-induced lipid peroxidation in spinal cord: protection with mannitol and methylprednisolone. J Free Radic Biol Med. 1985;1(1):59-64.
15. Hall ED, Yonkers PA, Tayler BM. Lack of effect of post injury treatment with MP or tirilared mesylate on the increase in eicosanoid levels in the aculely injured cat spinal cord. J Neuro Trauma, 1995;12(3):245.
16. Akdemir H, Pasaoglu A, Ozturk F, et al. Histopathology of experimental spinal cord trauma . Comparison of treatment with TRH , naloxone and dexamethasone. Res Exp Med Bed, 1992, 192 (2) : 177-183.
17. Pitts LH, Ross A, Chase GA, et al. Treatment with thyrotropin-releasing hormone (TRH) in patients with traumatic spinal cord injuries. J Neurotrauma. 1995 Jun;12(3):235-43.
18. Faden AI, Jacobs TP, Smith MT. Evaluation of the calcium channel antagonist nimodipine in experimental spinal cord ischemia. J Neurosurg. 1984 Apr;60(4):796-9.
19．赵凤东，王坤正，范顺武等．5-HT免疫组化染色观察尼莫地平于脊髓损伤中的保护作用．中国矫形外科杂志 2000年，12(7)
20. Gambardella G, Collufio D, Caruso GN, et al. Experimental incomplete spinal cord injury: treatment with a combination of nimodipine and adrenaline. J Neurosurg Sci. 1995 Mar;39(1):67-74.
21. Garcia,-M;Forster,-V;Hicks,-Detal.In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcineretina in vitro. Invest- Ophthalmol-Vis-Sci. 2003;44(10):4532～4541
22. Shimode,-H;Ueki,-A;Morita,-Y. Nerve growth factor attenuate ship pocampal choliner gicdeficits and operant learning impairment in rats with entorhinal cortex lesions. Behav-Pharmacol. 2003;14(3):179～190
23. Constantini S, Young W. The effects of methylprednisolone and the ganglioside GMlon acute spinalcord injury in rats. J Neurosurg, 1994, 80(1): 97-111.
24. Geisler FH, Dorsey FC, Coleman WP. GM-1 ganglioside in human spinal cord injury[J]. J of Neurotrauma,1992,9:s517-s530.
25．刘晓光，刘忠军，党耕町等．单唾液酸神经节甙脂(GM-1)治疗急性脊髓损伤的对比研究．中国脊柱脊髓杂志，2002，12(5)：344-347．
26. TatorCH.Strategies for recovery and regeneration after brain and spinal cord injury[J]. Injury Prev, 2002,8(24):33-36.
27.OConnorP. Incidence and patterns of spinal cord injury in Australia[J]. AccidAnalPrev, 2002,34(4):405-415.
28.BurkeDA, LindenRD, ZhangYP,etal. Epidemiology of spinal cord injury: a regional study[J]. SpinCord,2001,39(5):274-278.
29.ThompsonN, ShortD. The changing role of private funders in spinal cord injury reseach[J]. Spine,2001,26(24S):S24-26.
30.SekhonLHS, FehlingsMG Epidemiology, demographic and patho physiology of acute spinal cord injury[J]. Spine,2001,26(24S):S2-12.