瓦勒变性的调控暨对大鼠坐骨神经再生的影响研究
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摘要
周围神经损伤后,伤口处和远侧段全长神经纤维发生瓦勒变性,由于部分神经元凋亡,局部疤痕增生,吻合口处感觉、运动神经纤维轴突易相互错长,长期失神经支配后终末器官萎缩、变性等有害因素,导致治疗后功能恢复多数并不理想。因此神经损伤后的形态学变化和各种促进神经再生及功能恢复的方法成为近年来研究的热点。
目的:为了研究不同程度瓦勒变性对大鼠坐骨神经再生与功能恢复的影响及白细胞介素-1β对神经再生的影响机制,以及神经损伤后变性与再生过程的形态学变化规律,探索周围神经再生过程的可视化研究方法,为周围神经的治疗提供理论依据。
材料与方法:本课题第一部分研究在大鼠坐骨神经切割伤模型中分别利用白细胞介素-1β(IL-1β)和二氯亚甲基二磷酸盐脂质体(Cl2MDP)干预神经变性过程,损伤后1、3、5、7、15、30、60、90天用组织学方法、免疫组化方法检测损伤后不同时间点坐骨神经断端巨噬细胞的聚集与瓦勒变性程度,结合坐骨神经功能指数(SFI)、神经电生理等指标的恢复情况,评估不同程度的瓦勒变性对神经再生的影响。课题第二部分利用免疫组化方法检测损伤局部神经生长因子及胶质细胞源性神经营养因子的动态表达变化,结合组织学方法及轴突图像分析,探索白细胞介素-1β在神经损伤与再生中的作用机制。课题第三部分对不同时间点、不同部位的坐骨神经进行连续切片,经过改良三色染色法染色后光学显微镜观察并摄像后输入微机进行图像配准和二维处理,再利用图形工作站对坐骨神经损伤后变性、再生过程中典型的显微形态结构进行三维重建和显示,探讨坐骨神经再生过程的形态特征及量化规律。
结果:1. 伤后3天、7天硅胶管内可见淡红色胶冻样物质,组织易碎;伤后15天白细胞介素-1β处理组可见纤细浅红色条索状物把坐骨神经两断端连接起来;伤后30～90天各组硅胶管内再生神经逐渐变粗、变韧,颜色呈亮白色,但是白细胞介素-1β处理组再生神经直径明显大于Cl2MDP处理组和对照组,而后两组没有显著差异。
2. 光镜及电镜结果显示,IL-1β处理组神经损伤后巨噬细胞聚集的时间及轴突断裂、髓鞘崩解的时间均较早,变性组织清除也相应较快,增生的胶原纤维较少;再生神经轴突直径、数量及髓鞘厚度均优于对照组。Cl2MDP处理组早期很少有巨噬细胞出现于神经断端,变性组织清除较慢,再生神经分布密度也较小。
3.坐骨神经功能指数测定结果显示,白细胞介素-1β处理组坐骨神经功能指数恢复较快而且较好,Cl2MDP处理组早期恢复不如对照组,但后期与对照组差异不显著。
4.神经电生理结果显示,白细胞介素-1β处理组神经传导速度及复合肌肉动作电位波幅的恢复明显优于其余两组(P<0.01);Cl2MDP处理组复合肌肉动作电位波幅较对照组差(P<0.01),神经传导速度与对照组相比早期较差(P<0.01),晚期则无明显差异(P>0.05)。
5.神经损伤早期,白细胞介素-1β处理组损伤局部神经生长因子及胶质细胞源性神经营养因子的表达明显高于同一时期的对照组(P<0.01),神经再生晚期无明显差异性。
6.再生神经轴突图像分析结果显示,白细胞介素-1β处理组再生神经的有髓神经纤维的数目和轴突直径、髓鞘厚度均明显优于对照组(P<0.01)。
7.利用改良三色法对正常及再生神经进行染色并于光镜下摄像,输入微机进行图像配准和二维处理,在图形工作站中进行三维重建和显示,三维图像显示神经变性和再生过程较为典型的病理结构,如Bungner带的形成,郎飞氏节的结构等。
结论:1. 白细胞介素-1β可加剧神经损伤后瓦勒变性,使其变性组织清除加快,神经再生的程度及功能恢复较好。二氯亚甲基二磷酸盐脂质体耗竭了大鼠的单核细胞后,致使神经损伤后的变性组织清除延迟,早期神经再生较差,但对晚期神经功能恢复的影响较小。
2. 白细胞介素-1β能增加损伤早期局部的神经营养因子(NGF及GDNF)的合成与分泌,抑制胶质瘢痕的增生,对周围神经再生有一定的促进作用,再生小室内用药结合局部注射白细胞介素-1β可以作为治疗周围神经损伤的一种方法与手段。
3. 利用计算机图像重建技术,采用三维图像的形式对外周神经再生过程和再生规律进行可视化研究,是一种有效的方法。
The peripheral nerve injury is always accompanied by Wallerian degeneration in the wound and distal regions of the nerve and apoptosis of some related neurons.But some harmful factors such as wrong growth each other of motor and sensory neuritis in regeneration regions, the fibroplasias in nerve gap and atrophy of axon terminals after a long time of denervation always result the nerval function recovery is always dissatisfactory.So, it has become focus in clinical and basic research to investigate morphological changes of the injured nerve and measures of promoting peripheral nerve regeneration.
Objective
1) To study the effects of different extent of the wallerian degeneration on injured sciatic nerve regeneration and recovery of neural function in rat .
2) To investigate the mechanisms and effects of Interleukin-1 beta on nerve regeneration.
3) To research morphological changes of degenerating and regenerating sciatic nerve following injury, and grope for medical image visualization methods for peripheral nerve regeneration.The findings of these researches may provide theoretical basis for clinical therapy to peripheral nerve injury.
Methods
In the part one of the research, the rats' sciatic nerve was transected and then bridge-connected with silicone tube, The wallerian degeneration of transected sciatic nerve were intervened with recombinative human Interleukin-1 beta(IL-1β) and dichloromethylene diphosphonate-containing liposome(Cl2MDP) in respectively. Macrophage congregation in the local and the extent of wallerian degeneration were observed by pathological and immunohistochemical means on the 1、3、5、7、15、30、
60、90 days after injury, and the outcome of sciatic nerve function index(SFI)and Electrophysiolgical assessmentwere observed, by which the effects of different extent of wallerian degeneration on the nerve regeneration was evaluated.
In the part two of the research, the dynamic expression of the nerve growth factor (NGF)and Glial cell line-derived neurotrophic factor(GDNF) in the local were inspected with immunohistochemical technique, associating with pathology and morphometric analysis, the mechanisms and effects of Interleukin-1 beta were explored on injured peripheral nerve in rats.
In the part three of the research, The images of serial slices which were stained with trichrome stain was taken by micro-photograph system and inputted into computer by which the image matching and 2-D integrate processing were completed.The 3-D reconstruction and display of typical morphological structure of the injured nerve was accomplished in SGI workstation, and then we can explore morphological character of sciatic nerve regeneration and quantization method for nerve regeneration process.
Result
1.Soft red crisp colloid was seen in the silicone tube for 3 and 7 days after injury.15 days later it was observed that a soft red strip has connected two sides of transected sciatic nerve in the IL-1β group.After 30 to 90 days all of the newborn nerve became thicker , more tenacious, and its color became brighter than before, whereas, the diameter of the rebirth sciatic nerve in the IL-1β group were bigger than that of Cl2MDP and control groups.But the difference between the Cl2MDP group and control group has been inapparent .
2.The findings in the light microscope and electron microscope indicated that the time of macrophage assemble in the local region of injury and the time of axon rupture and myelin collapse in the IL-1β group was earlier than that of other groups, and the cleanness of degenerative tissue was more faster in the early stage in the IL-1β group.There was fewer hyperplastic collagen fiber in the IL-1β group.The indexes of regeneration nerve were superior to that of control group.On the contrary, there is fewer macrophage which invade into the local region in the early stage in the Cl2MDP group, and the elimination of degenerative tissue was also slow.The distributing density of the newborn nerve also was inferior.
3.The recovery rate of sciatic nerve functional inde
目的:为了研究不同程度瓦勒变性对大鼠坐骨神经再生与功能恢复的影响及白细胞介素-1β对神经再生的影响机制,以及神经损伤后变性与再生过程的形态学变化规律,探索周围神经再生过程的可视化研究方法,为周围神经的治疗提供理论依据。
材料与方法:本课题第一部分研究在大鼠坐骨神经切割伤模型中分别利用白细胞介素-1β(IL-1β)和二氯亚甲基二磷酸盐脂质体(Cl2MDP)干预神经变性过程,损伤后1、3、5、7、15、30、60、90天用组织学方法、免疫组化方法检测损伤后不同时间点坐骨神经断端巨噬细胞的聚集与瓦勒变性程度,结合坐骨神经功能指数(SFI)、神经电生理等指标的恢复情况,评估不同程度的瓦勒变性对神经再生的影响。课题第二部分利用免疫组化方法检测损伤局部神经生长因子及胶质细胞源性神经营养因子的动态表达变化,结合组织学方法及轴突图像分析,探索白细胞介素-1β在神经损伤与再生中的作用机制。课题第三部分对不同时间点、不同部位的坐骨神经进行连续切片,经过改良三色染色法染色后光学显微镜观察并摄像后输入微机进行图像配准和二维处理,再利用图形工作站对坐骨神经损伤后变性、再生过程中典型的显微形态结构进行三维重建和显示,探讨坐骨神经再生过程的形态特征及量化规律。
结果:1. 伤后3天、7天硅胶管内可见淡红色胶冻样物质,组织易碎;伤后15天白细胞介素-1β处理组可见纤细浅红色条索状物把坐骨神经两断端连接起来;伤后30～90天各组硅胶管内再生神经逐渐变粗、变韧,颜色呈亮白色,但是白细胞介素-1β处理组再生神经直径明显大于Cl2MDP处理组和对照组,而后两组没有显著差异。
2. 光镜及电镜结果显示,IL-1β处理组神经损伤后巨噬细胞聚集的时间及轴突断裂、髓鞘崩解的时间均较早,变性组织清除也相应较快,增生的胶原纤维较少;再生神经轴突直径、数量及髓鞘厚度均优于对照组。Cl2MDP处理组早期很少有巨噬细胞出现于神经断端,变性组织清除较慢,再生神经分布密度也较小。
3.坐骨神经功能指数测定结果显示,白细胞介素-1β处理组坐骨神经功能指数恢复较快而且较好,Cl2MDP处理组早期恢复不如对照组,但后期与对照组差异不显著。
4.神经电生理结果显示,白细胞介素-1β处理组神经传导速度及复合肌肉动作电位波幅的恢复明显优于其余两组(P<0.01);Cl2MDP处理组复合肌肉动作电位波幅较对照组差(P<0.01),神经传导速度与对照组相比早期较差(P<0.01),晚期则无明显差异(P>0.05)。
5.神经损伤早期,白细胞介素-1β处理组损伤局部神经生长因子及胶质细胞源性神经营养因子的表达明显高于同一时期的对照组(P<0.01),神经再生晚期无明显差异性。
6.再生神经轴突图像分析结果显示,白细胞介素-1β处理组再生神经的有髓神经纤维的数目和轴突直径、髓鞘厚度均明显优于对照组(P<0.01)。
7.利用改良三色法对正常及再生神经进行染色并于光镜下摄像,输入微机进行图像配准和二维处理,在图形工作站中进行三维重建和显示,三维图像显示神经变性和再生过程较为典型的病理结构,如Bungner带的形成,郎飞氏节的结构等。
结论:1. 白细胞介素-1β可加剧神经损伤后瓦勒变性,使其变性组织清除加快,神经再生的程度及功能恢复较好。二氯亚甲基二磷酸盐脂质体耗竭了大鼠的单核细胞后,致使神经损伤后的变性组织清除延迟,早期神经再生较差,但对晚期神经功能恢复的影响较小。
2. 白细胞介素-1β能增加损伤早期局部的神经营养因子(NGF及GDNF)的合成与分泌,抑制胶质瘢痕的增生,对周围神经再生有一定的促进作用,再生小室内用药结合局部注射白细胞介素-1β可以作为治疗周围神经损伤的一种方法与手段。
3. 利用计算机图像重建技术,采用三维图像的形式对外周神经再生过程和再生规律进行可视化研究,是一种有效的方法。
The peripheral nerve injury is always accompanied by Wallerian degeneration in the wound and distal regions of the nerve and apoptosis of some related neurons.But some harmful factors such as wrong growth each other of motor and sensory neuritis in regeneration regions, the fibroplasias in nerve gap and atrophy of axon terminals after a long time of denervation always result the nerval function recovery is always dissatisfactory.So, it has become focus in clinical and basic research to investigate morphological changes of the injured nerve and measures of promoting peripheral nerve regeneration.
Objective
1) To study the effects of different extent of the wallerian degeneration on injured sciatic nerve regeneration and recovery of neural function in rat .
2) To investigate the mechanisms and effects of Interleukin-1 beta on nerve regeneration.
3) To research morphological changes of degenerating and regenerating sciatic nerve following injury, and grope for medical image visualization methods for peripheral nerve regeneration.The findings of these researches may provide theoretical basis for clinical therapy to peripheral nerve injury.
Methods
In the part one of the research, the rats' sciatic nerve was transected and then bridge-connected with silicone tube, The wallerian degeneration of transected sciatic nerve were intervened with recombinative human Interleukin-1 beta(IL-1β) and dichloromethylene diphosphonate-containing liposome(Cl2MDP) in respectively. Macrophage congregation in the local and the extent of wallerian degeneration were observed by pathological and immunohistochemical means on the 1、3、5、7、15、30、
60、90 days after injury, and the outcome of sciatic nerve function index(SFI)and Electrophysiolgical assessmentwere observed, by which the effects of different extent of wallerian degeneration on the nerve regeneration was evaluated.
In the part two of the research, the dynamic expression of the nerve growth factor (NGF)and Glial cell line-derived neurotrophic factor(GDNF) in the local were inspected with immunohistochemical technique, associating with pathology and morphometric analysis, the mechanisms and effects of Interleukin-1 beta were explored on injured peripheral nerve in rats.
In the part three of the research, The images of serial slices which were stained with trichrome stain was taken by micro-photograph system and inputted into computer by which the image matching and 2-D integrate processing were completed.The 3-D reconstruction and display of typical morphological structure of the injured nerve was accomplished in SGI workstation, and then we can explore morphological character of sciatic nerve regeneration and quantization method for nerve regeneration process.
Result
1.Soft red crisp colloid was seen in the silicone tube for 3 and 7 days after injury.15 days later it was observed that a soft red strip has connected two sides of transected sciatic nerve in the IL-1β group.After 30 to 90 days all of the newborn nerve became thicker , more tenacious, and its color became brighter than before, whereas, the diameter of the rebirth sciatic nerve in the IL-1β group were bigger than that of Cl2MDP and control groups.But the difference between the Cl2MDP group and control group has been inapparent .
2.The findings in the light microscope and electron microscope indicated that the time of macrophage assemble in the local region of injury and the time of axon rupture and myelin collapse in the IL-1β group was earlier than that of other groups, and the cleanness of degenerative tissue was more faster in the early stage in the IL-1β group.There was fewer hyperplastic collagen fiber in the IL-1β group.The indexes of regeneration nerve were superior to that of control group.On the contrary, there is fewer macrophage which invade into the local region in the early stage in the Cl2MDP group, and the elimination of degenerative tissue was also slow.The distributing density of the newborn nerve also was inferior.
3.The recovery rate of sciatic nerve functional inde
引文
Meyer R,Clauissen GC,Oh SJ. Modified trichrome staining technique of the nerve to determine proximal nerve viability. Microsurgery. 1995,16:129-132
朱长庚.神经解剖学.北京:人民卫生出版社,2002,3-112
Cajal's,Regeneration and regeneration of the nervous system (translated by May R M).New York,Oxford: Oxford University Press ,1991
Lubinska L.Patterns of Wallerian degeneration of myelinated fibres in short and long peripheral stumps and in isolated segments of rat phrenic nerve.Interpretation of the role of axoplasmic flow of the trophic factor. Brain Res,1982,233:227-240
范惠勇,郑德枢,等.周围神经Waller变性时信息传递方式,神经解剖杂志,2001,17(3),205-208.
Ide C, Tohyama K, Yokota R, et al. Schwann cell basal lamina and nerve regeneration. Brain Res,1983,288: 61-75
Wang G Y,Hirai K, Shimada H.The role of laminin, a component of Schwann cell basal lamina, in rat sciatic nerve regeneration within antiserum-treated nerve grafts.Brain Res. 1992,570: 116-125
Bigbee J W, Yoshino J E,DeVries G H.Morphologic and proliferative responses of cultured Schwann cells following rapid phagocytosis of a myelin-enriched fracrion.J Neurocytol,1987,16:487-496
Beuche W. Friede R L.The role of non-resident cells in Wallerian degeneration. J Neurocytol,1984,13: 767-796
Stoll G, Griffin J W, Li C Y. Wallerian degeneration in the Peripheral nervous system:Participation of both Schwann cells and macrophages in myelin degeneration.J Neurocytol,1989,18,671-683
范惠勇,郑德枢.周围神经Waller变性时巨噬细胞来源的探讨.神经解剖学杂志. 2002 .18(2):131～134
刘香梅.神经生长的生物学和病理学.北京:科学出版社,1985:170-191.
Perry V H,Brown M C. Role of macrophages in peripheral nerve degeneration and repair.Bioessays,1992,14: 401-406
Gregson N A,Hall S M. A quantitative analysis of the effects of the Intraneural injection
of lysophosphatidyl choline. J-Cell-Sci,1973,13: 257-277
Paul J A,Gregson N A.An immunohistochemical study of phospholipase A2 in peripheral nerve during Wallerian degeneration.J Neuroimmmunol,1992,39:31-48
Shimizu T. Wolfe L S. Arachidonic acid cascade and signal transduction. J Neurochem,1990,55: 1-13
Trotter J,smith M E. The role of phospholipase from inflammatory macrophages in demyelination .Neurochem Res. 1986: 11:349-361
Battisti W P, Wang J, Bozek K,at el. Macrophages, microglia, and astrocytes are rapidly activated after crush injury of the goldfish optic nerve: a light and electron microscopic analysis. J-Comp-Neurol,1995,354(2): 306-320
Hirata K,Mitoma H,Ueno N,et al.Differential response of macrophage subpopulations to myelin degradation in the injured rat sciatic nerve. J Neurocytol,1999,28:685-695
Bruck W,Huitinga I,Dijkstra CD,Liposome-mediated monocyte depletion during wallerian degeneration defines the role of hematogenous phagocytes in myelin removal. J-Neurosci-Res. 1996 Nov 15,4(4):477-484
Bruck W,Friede R L. Anti-macrophage CR3 antibody blocks myelin phagocytosis by macrophages in vitro.Acta-Neuropathol-(Berl).1990,80:415-418
Heumann R,Lindholm D,Bandtlow C,et al.Differential regulation of mRNA encoding nerve growth factor and its receptor in rat sciatic nerve during development degeneration and regeneration: Role of macrophages. Proc Natl Acad Sci USA, 1987, 84:8735-8739
Lindholm D, Heumann R, Meyer M, et al. Interleukin-1 regulates synthesis of nerve growth factor hi non-neuronal cells of rat sciatic nerve. Nature, 1987,330:658-659
Nathan CF. Secretory products of macrophages. J Clin Invest, 1987,79:319-326
Lindholm D, Heumann R, Hengerer B, et al. Interleukin-1 increases stability and transcription of mRNA encoding nerve growth factor in cultured rat fibroblast. J Biol Chem, 1988,263:16348-16351
Guenard V, Dinarello CA, Weston PJ, et al. Peripheral nerve regeneration is impeded by interleukin-1 receptor antagonist released from a polymeric guidance channel. J Neurosci Res, 1991,29:396-400
Van Rooijen ,The liposome-mediated macrophage `suicide' technique, Journal of
Immunological Methods, 1989,124(1):1-6
黄涛,秦建强,熊绍虎,余磊,等。周围神经损伤后巨噬细胞游走抑制因子mRNA在雪旺细胞的表达状况.中华外科杂志.2002 40:699-701
Carroll S L, Frohnert P W.Expression of JE(monocyte chemoattractant protein 1)is induced by sciatic axotomy in wild type rodents but not in C57BL/WLD(s) mice. J Neuropathol Exp Neurol.1998.57:915-930
黄涛,秦建强,刘大庸,等.外周神经损伤后雪旺细胞激活巨噬细胞的实验研究.中华显微外科杂志.2002 .25:192-193
Toews A D, Barrett C, Morell P. Monocyte chemoattractant protein-1 is responsible for macrophage recruitment following injury to sciatic nerve. J Neurosci Res.1998.53:260-267
Cammer W, Bloom BR, et al. Degradation of basic protein in myelin by neutral proteases secreted by stimulated macrophages: a possible mechanism of inflammatory demyelination.Proc Natl Acad sci USA, 1978: 75 (3):1664
Warner SJC,Auger BR, et al. Human interleukin-1 induces interleukin-1 gene expression in human vascular smooth muscle ce11s.J Exp Med,1987.165:1316
Dinarello CAT, lkejima SJC, et al. Interleukin-1 induces interleukin-1. Induction of circulating interleukin-1 in rabbits in vivo and in huuan mononuclear cells in vitro.J lmmunol, 1987. 139:1902
Salzer JL, Bunge RP, Glaser L. Studies of Schwann cell proliferation: Evidence for the surface localization of the neurite mitogen. J Cell Biol,1980,84:767-778
Salzer J L, Williams A K, Glaser L, et al. Studies of Schwann cell proliferation Characterization of the stimulation and specificity of the response to a neurite membrane fraction. J Cell Biol, 1980.84:753-766
Goodrum JF, Fow ler KA, H ostettler JD, et al. Peripheral nerve regeneration and cholesterol reutilization are normal in the low-density lipoprotein receptor knockout mouse.J Neurosci Res, 2000, 59: 581-586
杨静,裴福兴,杨志明等.IL-1促周围神经再生的实验研究.中华创伤杂志.1998.14(2):77-79
Yasuda T,Sobue G,Ito T,etal.Nerve growth factor enhance neurite arborization of adult sensory neurons: a study in single-cell culture.Brain Reseurch.1990.524 (1): 54-63
Gold B G,Mobley W C, Matheson S F.Regulation of axonal caliber, neurofilament content, and nuclear localization in mature sensory neurons by nerve growth factor.J Neurosci.1991.11(4): 943-955.
Derby A, Engleman VW, Frierdich GE, et al. Nerve growth factor facilitates regeneration across nerve gaps, Morphological and behavioral studies in rat sciatic nerve. Exp Neurol, 1993.119:176-191
Derby A, Engleman VW, Neises G, et al. Exogenous nerve growth factor enhances the regeneration of rat sciatic nerve. Soc Neurosci Abstr, 1988.14:498-503
Rich KM, Alexander TD, Pryoor JC, et al. Nerve growth factor enhances regeneration through silicone chambers.Exp Neurol,1989 , 105(2): 162-170
尹方明,秦建强等,P75在周围神经损伤后的表达. 2000. 18 (1):71-72
Pezet S, Onteniente B, Jullien J,et al.Differential regulation of NGF receptors in primary sensory neurons by adjuvant-induced arthritis in the rat.Pain,2001, 90(1-2):113-125.
Rush RA. Immunohistochemical localization of endogenous nerve growth factor. Nature, 1984.312,364-367
Taniuchi M, Clark HB, Johnson EM. Induction of nerve growth factor receptors in Schwann cells after axotomy. AProc Natl Acad Sci USA, 1986.83,4094-4098
许家军,何成,宋田斌,等.周围神经损伤后外源性神经生长因子及睫状神经营养因子对神经元的保护作用. 中华显微外科杂志,1997,20(1):32-35
Korsching S, Thoenen H. Treatment with 6-hydroxydopamine and colchicine decreases nerve growth factor levels in sympathetic ganglia and increase them in the corresponding target tissue. J Neurosci, 1985.5:1058-1061
Heumann R, Koraching S, Bandtlow C, et al. Nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transection.J Cell Biol, 1987,104:1623-1631
Heumann R, Schwab ME, et al. Cellular localization of nerve growth factor synthesis by in situ hybridization. EMBO J, 1987.6:891-899
LARS B D.Stimulation of regeneration of the sciatic never by experimentally induced inflammation in rats [J] .Scand J Plast Reconstr Hand Surg, 1992, 26:121-125.
Lin LF,Doherty DH,Lile JD et al .GDNF:a glial cell line-derived neurotrophic factor
for midbrain dopaminergic neurons.Science,1993,260 (5111):1130-1132.
Henderson CE,Philips HS,Pollock RA, et al .GDNF:A potent survival factor for motoneurons present in peripheral nerve and muscle .Science,1994,266:1062-1064.
Itupp M, Ryden M, Jornvall H et al .Peripheral expression and biological activities GDNF:A new neurotrophic factor from avian and mammalian peripheral neurons .J Cell Biol, 1995,130(1):137-148.
Sanicola M, Hession C,Worley D et al.Glial cell line-derived neurotrophic factor dependent RET activation can be mediated by two different cell-surface accessory proteins .Proc Natl A cad Sci USA, 1997,94:6238-6243.
Yan Q,Wang J,Matheson CR,et al.Glial cell line-derived neurotrophic factor(GDNF) promotes the survival of axotomized retinal ganglion cells in adult rats:comparison to and combination with brain derived neurotrophic factor(BDNF). J Neurobiol,1999,38(3):382-390.
Walton KM. GDNF:a novel factor with therapeutic potential for neurodegenerative disorders.Nbl Neurobiol,1999,19 (1):43-59.
宋海涛,贾连顺,陈坚,等.坐骨神经切断后GDNF mRNA在两侧断端的表达变化.中华骨科杂志.2002,22(3):176-178
Appel E, Kolman O, Kazimirsky G,etal.Regulation of GDNF expression in cultured astrocytes by inflammatory stimuli.NeuroReport , 1997,8: 3309–3312
陈哲宇,曹莉,路长林,等.周围神经损伤后外源性GDNF对神经元的保护作用.生理学报,2000,52(4):295～300
陈振兵,洪光祥,王发斌,等.鼠神经胶原纤维的免疫组织化学与电镜观察.中华手外科杂志.1997,13(1):53-54
Bhatnagar R.Interleukin-1 inhibits the synthesis of collagen by fibroblasts.Biochem Internat,1986,13:709-711
Arturo. Regulation of human lung fibroblast aj I) procollagen gene expression by TNT, IL-1 p ,and PGF2 .J Biolog Chem,1993 ,268:10364-10366
杨东元,罗锦辉,朱金华.白细胞介素-1对病理性瘢痕成纤维细胞胶原合成的影响. 中华整形烧伤外科杂志.1999, 15(6):434-436
Nicole A,Sprewings K.Effect of PGF2 on proline uptake and protein synthesis by cultured human mesangial cells . Kidney International ,1990 ,38 :1151-1153
Ronald H. The Effext of PGF2 on amine acid uptake and protein formation by lung fibroblasts.J Biolog Chem,1986,261:8734-8137
Bruce J.Mandel K.Effect of cAMP on the intracellular degradation of newly synthesized collagen .J Biolog Chem,1980,255:2843-2848
Arnold E.IL-1 stimulati on of collage nase production by cultured fibroblasts.J Experim Med,1983,157:801-805
Mrosovky N,Molony LA, Conn CA, et al.Anorexic effects of interleukin-1 in the rat. Am .J. Physiol 1989, 257: 1315-1321.
Dunlap KD, Church DR.Interleukin-1 beta reduces daily activity level in male lizards, scelopores occidentalis. Brain Behav Immun 1996,10(1):68-73.
邓平,宋一璇,祝家镇.改良三色法显示石蜡切片的神经轴突及神经髓鞘.中华病理学杂志.1998, 3:235
罗述谦.医学图像配准技术.国外医学·生物医学工程分册,1999, 22(1):1-7
秦斌杰,陈旭,庄天戈.医学图像三维可视化.航天医学与医学工程.2001,6:452
Otten E, van Lecuwen JL. A 3-D reconstruction package from serial section images.Eur J Cell Biol,1989,48(25):73
王伯沄.病理学技术.第一版.北京:人民卫生出版社,2000:68-75
朱长庚.神经解剖学.北京:人民卫生出版社,2002,3-112
Cajal's,Regeneration and regeneration of the nervous system (translated by May R M).New York,Oxford: Oxford University Press ,1991
Lubinska L.Patterns of Wallerian degeneration of myelinated fibres in short and long peripheral stumps and in isolated segments of rat phrenic nerve.Interpretation of the role of axoplasmic flow of the trophic factor. Brain Res,1982,233:227-240
范惠勇,郑德枢,等.周围神经Waller变性时信息传递方式,神经解剖杂志,2001,17(3),205-208.
Ide C, Tohyama K, Yokota R, et al. Schwann cell basal lamina and nerve regeneration. Brain Res,1983,288: 61-75
Wang G Y,Hirai K, Shimada H.The role of laminin, a component of Schwann cell basal lamina, in rat sciatic nerve regeneration within antiserum-treated nerve grafts.Brain Res. 1992,570: 116-125
Bigbee J W, Yoshino J E,DeVries G H.Morphologic and proliferative responses of cultured Schwann cells following rapid phagocytosis of a myelin-enriched fracrion.J Neurocytol,1987,16:487-496
Beuche W. Friede R L.The role of non-resident cells in Wallerian degeneration. J Neurocytol,1984,13: 767-796
Stoll G, Griffin J W, Li C Y. Wallerian degeneration in the Peripheral nervous system:Participation of both Schwann cells and macrophages in myelin degeneration.J Neurocytol,1989,18,671-683
范惠勇,郑德枢.周围神经Waller变性时巨噬细胞来源的探讨.神经解剖学杂志. 2002 .18(2):131～134
刘香梅.神经生长的生物学和病理学.北京:科学出版社,1985:170-191.
Perry V H,Brown M C. Role of macrophages in peripheral nerve degeneration and repair.Bioessays,1992,14: 401-406
Gregson N A,Hall S M. A quantitative analysis of the effects of the Intraneural injection
of lysophosphatidyl choline. J-Cell-Sci,1973,13: 257-277
Paul J A,Gregson N A.An immunohistochemical study of phospholipase A2 in peripheral nerve during Wallerian degeneration.J Neuroimmmunol,1992,39:31-48
Shimizu T. Wolfe L S. Arachidonic acid cascade and signal transduction. J Neurochem,1990,55: 1-13
Trotter J,smith M E. The role of phospholipase from inflammatory macrophages in demyelination .Neurochem Res. 1986: 11:349-361
Battisti W P, Wang J, Bozek K,at el. Macrophages, microglia, and astrocytes are rapidly activated after crush injury of the goldfish optic nerve: a light and electron microscopic analysis. J-Comp-Neurol,1995,354(2): 306-320
Hirata K,Mitoma H,Ueno N,et al.Differential response of macrophage subpopulations to myelin degradation in the injured rat sciatic nerve. J Neurocytol,1999,28:685-695
Bruck W,Huitinga I,Dijkstra CD,Liposome-mediated monocyte depletion during wallerian degeneration defines the role of hematogenous phagocytes in myelin removal. J-Neurosci-Res. 1996 Nov 15,4(4):477-484
Bruck W,Friede R L. Anti-macrophage CR3 antibody blocks myelin phagocytosis by macrophages in vitro.Acta-Neuropathol-(Berl).1990,80:415-418
Heumann R,Lindholm D,Bandtlow C,et al.Differential regulation of mRNA encoding nerve growth factor and its receptor in rat sciatic nerve during development degeneration and regeneration: Role of macrophages. Proc Natl Acad Sci USA, 1987, 84:8735-8739
Lindholm D, Heumann R, Meyer M, et al. Interleukin-1 regulates synthesis of nerve growth factor hi non-neuronal cells of rat sciatic nerve. Nature, 1987,330:658-659
Nathan CF. Secretory products of macrophages. J Clin Invest, 1987,79:319-326
Lindholm D, Heumann R, Hengerer B, et al. Interleukin-1 increases stability and transcription of mRNA encoding nerve growth factor in cultured rat fibroblast. J Biol Chem, 1988,263:16348-16351
Guenard V, Dinarello CA, Weston PJ, et al. Peripheral nerve regeneration is impeded by interleukin-1 receptor antagonist released from a polymeric guidance channel. J Neurosci Res, 1991,29:396-400
Van Rooijen ,The liposome-mediated macrophage `suicide' technique, Journal of
Immunological Methods, 1989,124(1):1-6
黄涛,秦建强,熊绍虎,余磊,等。周围神经损伤后巨噬细胞游走抑制因子mRNA在雪旺细胞的表达状况.中华外科杂志.2002 40:699-701
Carroll S L, Frohnert P W.Expression of JE(monocyte chemoattractant protein 1)is induced by sciatic axotomy in wild type rodents but not in C57BL/WLD(s) mice. J Neuropathol Exp Neurol.1998.57:915-930
黄涛,秦建强,刘大庸,等.外周神经损伤后雪旺细胞激活巨噬细胞的实验研究.中华显微外科杂志.2002 .25:192-193
Toews A D, Barrett C, Morell P. Monocyte chemoattractant protein-1 is responsible for macrophage recruitment following injury to sciatic nerve. J Neurosci Res.1998.53:260-267
Cammer W, Bloom BR, et al. Degradation of basic protein in myelin by neutral proteases secreted by stimulated macrophages: a possible mechanism of inflammatory demyelination.Proc Natl Acad sci USA, 1978: 75 (3):1664
Warner SJC,Auger BR, et al. Human interleukin-1 induces interleukin-1 gene expression in human vascular smooth muscle ce11s.J Exp Med,1987.165:1316
Dinarello CAT, lkejima SJC, et al. Interleukin-1 induces interleukin-1. Induction of circulating interleukin-1 in rabbits in vivo and in huuan mononuclear cells in vitro.J lmmunol, 1987. 139:1902
Salzer JL, Bunge RP, Glaser L. Studies of Schwann cell proliferation: Evidence for the surface localization of the neurite mitogen. J Cell Biol,1980,84:767-778
Salzer J L, Williams A K, Glaser L, et al. Studies of Schwann cell proliferation Characterization of the stimulation and specificity of the response to a neurite membrane fraction. J Cell Biol, 1980.84:753-766
Goodrum JF, Fow ler KA, H ostettler JD, et al. Peripheral nerve regeneration and cholesterol reutilization are normal in the low-density lipoprotein receptor knockout mouse.J Neurosci Res, 2000, 59: 581-586
杨静,裴福兴,杨志明等.IL-1促周围神经再生的实验研究.中华创伤杂志.1998.14(2):77-79
Yasuda T,Sobue G,Ito T,etal.Nerve growth factor enhance neurite arborization of adult sensory neurons: a study in single-cell culture.Brain Reseurch.1990.524 (1): 54-63
Gold B G,Mobley W C, Matheson S F.Regulation of axonal caliber, neurofilament content, and nuclear localization in mature sensory neurons by nerve growth factor.J Neurosci.1991.11(4): 943-955.
Derby A, Engleman VW, Frierdich GE, et al. Nerve growth factor facilitates regeneration across nerve gaps, Morphological and behavioral studies in rat sciatic nerve. Exp Neurol, 1993.119:176-191
Derby A, Engleman VW, Neises G, et al. Exogenous nerve growth factor enhances the regeneration of rat sciatic nerve. Soc Neurosci Abstr, 1988.14:498-503
Rich KM, Alexander TD, Pryoor JC, et al. Nerve growth factor enhances regeneration through silicone chambers.Exp Neurol,1989 , 105(2): 162-170
尹方明,秦建强等,P75在周围神经损伤后的表达. 2000. 18 (1):71-72
Pezet S, Onteniente B, Jullien J,et al.Differential regulation of NGF receptors in primary sensory neurons by adjuvant-induced arthritis in the rat.Pain,2001, 90(1-2):113-125.
Rush RA. Immunohistochemical localization of endogenous nerve growth factor. Nature, 1984.312,364-367
Taniuchi M, Clark HB, Johnson EM. Induction of nerve growth factor receptors in Schwann cells after axotomy. AProc Natl Acad Sci USA, 1986.83,4094-4098
许家军,何成,宋田斌,等.周围神经损伤后外源性神经生长因子及睫状神经营养因子对神经元的保护作用. 中华显微外科杂志,1997,20(1):32-35
Korsching S, Thoenen H. Treatment with 6-hydroxydopamine and colchicine decreases nerve growth factor levels in sympathetic ganglia and increase them in the corresponding target tissue. J Neurosci, 1985.5:1058-1061
Heumann R, Koraching S, Bandtlow C, et al. Nerve growth factor synthesis in nonneuronal cells in response to sciatic nerve transection.J Cell Biol, 1987,104:1623-1631
Heumann R, Schwab ME, et al. Cellular localization of nerve growth factor synthesis by in situ hybridization. EMBO J, 1987.6:891-899
LARS B D.Stimulation of regeneration of the sciatic never by experimentally induced inflammation in rats [J] .Scand J Plast Reconstr Hand Surg, 1992, 26:121-125.
Lin LF,Doherty DH,Lile JD et al .GDNF:a glial cell line-derived neurotrophic factor
for midbrain dopaminergic neurons.Science,1993,260 (5111):1130-1132.
Henderson CE,Philips HS,Pollock RA, et al .GDNF:A potent survival factor for motoneurons present in peripheral nerve and muscle .Science,1994,266:1062-1064.
Itupp M, Ryden M, Jornvall H et al .Peripheral expression and biological activities GDNF:A new neurotrophic factor from avian and mammalian peripheral neurons .J Cell Biol, 1995,130(1):137-148.
Sanicola M, Hession C,Worley D et al.Glial cell line-derived neurotrophic factor dependent RET activation can be mediated by two different cell-surface accessory proteins .Proc Natl A cad Sci USA, 1997,94:6238-6243.
Yan Q,Wang J,Matheson CR,et al.Glial cell line-derived neurotrophic factor(GDNF) promotes the survival of axotomized retinal ganglion cells in adult rats:comparison to and combination with brain derived neurotrophic factor(BDNF). J Neurobiol,1999,38(3):382-390.
Walton KM. GDNF:a novel factor with therapeutic potential for neurodegenerative disorders.Nbl Neurobiol,1999,19 (1):43-59.
宋海涛,贾连顺,陈坚,等.坐骨神经切断后GDNF mRNA在两侧断端的表达变化.中华骨科杂志.2002,22(3):176-178
Appel E, Kolman O, Kazimirsky G,etal.Regulation of GDNF expression in cultured astrocytes by inflammatory stimuli.NeuroReport , 1997,8: 3309–3312
陈哲宇,曹莉,路长林,等.周围神经损伤后外源性GDNF对神经元的保护作用.生理学报,2000,52(4):295～300
陈振兵,洪光祥,王发斌,等.鼠神经胶原纤维的免疫组织化学与电镜观察.中华手外科杂志.1997,13(1):53-54
Bhatnagar R.Interleukin-1 inhibits the synthesis of collagen by fibroblasts.Biochem Internat,1986,13:709-711
Arturo. Regulation of human lung fibroblast aj I) procollagen gene expression by TNT, IL-1 p ,and PGF2 .J Biolog Chem,1993 ,268:10364-10366
杨东元,罗锦辉,朱金华.白细胞介素-1对病理性瘢痕成纤维细胞胶原合成的影响. 中华整形烧伤外科杂志.1999, 15(6):434-436
Nicole A,Sprewings K.Effect of PGF2 on proline uptake and protein synthesis by cultured human mesangial cells . Kidney International ,1990 ,38 :1151-1153
Ronald H. The Effext of PGF2 on amine acid uptake and protein formation by lung fibroblasts.J Biolog Chem,1986,261:8734-8137
Bruce J.Mandel K.Effect of cAMP on the intracellular degradation of newly synthesized collagen .J Biolog Chem,1980,255:2843-2848
Arnold E.IL-1 stimulati on of collage nase production by cultured fibroblasts.J Experim Med,1983,157:801-805
Mrosovky N,Molony LA, Conn CA, et al.Anorexic effects of interleukin-1 in the rat. Am .J. Physiol 1989, 257: 1315-1321.
Dunlap KD, Church DR.Interleukin-1 beta reduces daily activity level in male lizards, scelopores occidentalis. Brain Behav Immun 1996,10(1):68-73.
邓平,宋一璇,祝家镇.改良三色法显示石蜡切片的神经轴突及神经髓鞘.中华病理学杂志.1998, 3:235
罗述谦.医学图像配准技术.国外医学·生物医学工程分册,1999, 22(1):1-7
秦斌杰,陈旭,庄天戈.医学图像三维可视化.航天医学与医学工程.2001,6:452
Otten E, van Lecuwen JL. A 3-D reconstruction package from serial section images.Eur J Cell Biol,1989,48(25):73
王伯沄.病理学技术.第一版.北京:人民卫生出版社,2000:68-75