脉冲电磁场影响大鼠坐骨神经损伤后功能恢复的实验研究
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摘要
背景:
周围神经(Peripheral nerve)损伤是临床的多发病、常见病,是人类致残的重要原因之一。不同原因引起的周围神经损伤,其损伤程度不同,导致临床表现不同,治疗方法与预后也大不相同。治疗方法包括药物治疗、物理治疗、手术治疗与细胞治疗等,但功能恢复仍不够理想。因此,如何有效的促进周围神经损伤后的再生和功能恢复仍是临床工作关注的热点。
物理治疗具有安全、无创、方便、有效等特点,日益受到临床医患的青睐。大量研究认为,脉冲电磁场具有增加骨密度、促进骨折愈合的作用,临床常运用于骨质疏松理疗。有零星报告脉冲电磁场也能促进受损周围神经的修复,但也有结果不支持的报道。因此,有必要进行相关实验研究,以明确其治疗的有效性与参数,并且探讨其作用机制,为拓展脉冲电磁场临床治疗周围神经损伤提供佐证。
材料与方法:
实验一按随机分组原则将SD大鼠30只分成5组:正常对照(A组);模型对照(B组); 0.3mT 2Hz (C组); 0.3mT 10Hz (D组); 0.3mT 40Hz (E组),每组6只。除正常对照组外,对所有大鼠按文献方法钳夹坐骨神经造模,造模成功后并定位标记。C、D、E组每天在强度为0.3mT,频率分别为2Hz、10Hz、40Hz的脉冲电磁场仪器中接受治疗,参考相关文献给予大鼠2h/次,2次/天,共30/d的治疗疗程,正常对照组与模型对照组不干预。测定坐骨神经功能指数(SFI)及神经传导速度(NCV),比较研究不同频率脉冲电磁场对大鼠坐骨神经损伤后功能恢复的影响。
实验二30只SD大鼠随机分成三组:模型对照(A组)、正常对照(B组)和治疗(C组),每组10只。除正常对照组不处理外,其余组按实验一方法造模。治疗组给予合适频率(根据实验一结果)的脉冲电磁场治疗,2h/次,2次/天,模型对照组和正常对照组不给予任何干预。于1,2,4周除正常对照组外,对其余两组按随机抽样原则分别取5只大鼠,取其离钳夹伤5mm处的坐骨神经进行透射电镜观察,及免疫组化检测。并于第4周对模型对照组与治疗组轴突图象分析。
结果:
在强度与干预时间相同的前提下,0.3mT 2Hz组大鼠SFI及NCV显著高于其他频率组(P<0.05)。2Hz脉冲电磁场处理大鼠损伤坐骨神经后, Wallerian变性进程加速,雪旺氏细胞的增殖明显增加,髓鞘再生明显活跃,损伤坐骨神经中NGF表达显著高于模型对照组(P<0.05)。
结论:
0.3mT 2Hz组较10、40Hz组更能促进大鼠坐骨神经损伤后的功能恢复。在组织形态学上,脉冲电磁场能促进雪旺氏细胞的增殖,髓鞘增生;在分子生物学上,能促进NGF表达等,这可能是脉冲电磁场促进周围神经损伤后再生和功能恢复的机制之一。
Background:
Peripheral nerve injury is a common disease which is a major reason for disability in clinical. The clinical performance and prognosis are different because of different reasons. Treatment methods include medication、physical therapy、surgical therapy and cell therapy. However, the functional recovery is not ideal. It is also an outstanding issue for nerve injury repairing in the clinical. How to promote the regeneration and functional recovery after nerve injury becomes a focus.
Physical therapy is safe、non-invasion、convenient and effective ,which is favored by the clinical. A large number of studies suggest that pulsed electromagnetic fields can increase bone mineral density, and promote the fracture healing. It is often used in treating the osteoporosis. What’s more, some reports said that pulsed electromagnetic fields can promote the peripheral nerve injury repairing, however, some do not support. Therefore, it is necessary to do the related experimental researches to make its effectiveness and the parameters clearly, also the mechanism. Providing the evidences in treating the peripheral nerve injury to realize the clinical values.
Methods and materials:
Experiment 1 According to the principle of randomized, 30 SD rats were divided into five groups: the normal control (group A), model control (group B), 0.3mT 2Hz (group C), 0.3mT 10Hz (group D), 0.3mT 40Hz (group E), each 6. Except the normal control group, the other rats were clamped into sciatic nerve injury model according to the literature. After finishing the model, we gave them location. C、D、E group accepted the treatment , the PEMF’s field strength was 0.3mT, the frequency was 2Hz, 10Hz, 40Hz respectively. 2h/time, 2times/day, 30days altogether according the literature. the normal control group and model control group gave no intervention. Determined the sciatic nerve function index (SFI) and nerve conduction velocity(NCV) to all rats.
Experiment 2 Select 30 SD rats, divided into 3 groups: model control(group A)、normal control (group B)and treatment (group C), each 10. Except the normal control group, the other rats were received clamped sciatic nerve model according to the experiment 1. The treatment group were received 0.3mT 2Hz PEMF (according to the experiment 1’s result),2h/time, 2times/day, model control group and normal control group were not given any treatment. Except the normal control group,we took 5 rats from the other two groups respectively,Then the nerves which was 5mm away from the clamped were sampled at the 1~(st), 2~(nd), 4~(th) week respectively, observed by electron microscope and immunohistoc-hemistry. Image pattern analysis of axon was done at the 4th week after operation to model control group and treatment group.
Results:
In the same strength and time, 2Hz group’s SFI and NCV were higher than other frequency groups significantly (P<0.05). 2Hz PEMF make the course of Wallerian degeneration accelerated、schwann’s cell proliferation increased significantly、regeneration of axon improved obviously. The NGFof the treatment group expressed more than the model control group (P<0.05).
Conclusion:
0.3mT 2Hz PEMF may be suitable for treating the sciatic nerve injury. one of the mechanism may be: PEMF accelerated the course of Wallerian degeneration,Schwann’s cell proliferation,regeneration of axon, and the expression of NGF in nerve’s fiber.
周围神经(Peripheral nerve)损伤是临床的多发病、常见病,是人类致残的重要原因之一。不同原因引起的周围神经损伤,其损伤程度不同,导致临床表现不同,治疗方法与预后也大不相同。治疗方法包括药物治疗、物理治疗、手术治疗与细胞治疗等,但功能恢复仍不够理想。因此,如何有效的促进周围神经损伤后的再生和功能恢复仍是临床工作关注的热点。
物理治疗具有安全、无创、方便、有效等特点,日益受到临床医患的青睐。大量研究认为,脉冲电磁场具有增加骨密度、促进骨折愈合的作用,临床常运用于骨质疏松理疗。有零星报告脉冲电磁场也能促进受损周围神经的修复,但也有结果不支持的报道。因此,有必要进行相关实验研究,以明确其治疗的有效性与参数,并且探讨其作用机制,为拓展脉冲电磁场临床治疗周围神经损伤提供佐证。
材料与方法:
实验一按随机分组原则将SD大鼠30只分成5组:正常对照(A组);模型对照(B组); 0.3mT 2Hz (C组); 0.3mT 10Hz (D组); 0.3mT 40Hz (E组),每组6只。除正常对照组外,对所有大鼠按文献方法钳夹坐骨神经造模,造模成功后并定位标记。C、D、E组每天在强度为0.3mT,频率分别为2Hz、10Hz、40Hz的脉冲电磁场仪器中接受治疗,参考相关文献给予大鼠2h/次,2次/天,共30/d的治疗疗程,正常对照组与模型对照组不干预。测定坐骨神经功能指数(SFI)及神经传导速度(NCV),比较研究不同频率脉冲电磁场对大鼠坐骨神经损伤后功能恢复的影响。
实验二30只SD大鼠随机分成三组:模型对照(A组)、正常对照(B组)和治疗(C组),每组10只。除正常对照组不处理外,其余组按实验一方法造模。治疗组给予合适频率(根据实验一结果)的脉冲电磁场治疗,2h/次,2次/天,模型对照组和正常对照组不给予任何干预。于1,2,4周除正常对照组外,对其余两组按随机抽样原则分别取5只大鼠,取其离钳夹伤5mm处的坐骨神经进行透射电镜观察,及免疫组化检测。并于第4周对模型对照组与治疗组轴突图象分析。
结果:
在强度与干预时间相同的前提下,0.3mT 2Hz组大鼠SFI及NCV显著高于其他频率组(P<0.05)。2Hz脉冲电磁场处理大鼠损伤坐骨神经后, Wallerian变性进程加速,雪旺氏细胞的增殖明显增加,髓鞘再生明显活跃,损伤坐骨神经中NGF表达显著高于模型对照组(P<0.05)。
结论:
0.3mT 2Hz组较10、40Hz组更能促进大鼠坐骨神经损伤后的功能恢复。在组织形态学上,脉冲电磁场能促进雪旺氏细胞的增殖,髓鞘增生;在分子生物学上,能促进NGF表达等,这可能是脉冲电磁场促进周围神经损伤后再生和功能恢复的机制之一。
Background:
Peripheral nerve injury is a common disease which is a major reason for disability in clinical. The clinical performance and prognosis are different because of different reasons. Treatment methods include medication、physical therapy、surgical therapy and cell therapy. However, the functional recovery is not ideal. It is also an outstanding issue for nerve injury repairing in the clinical. How to promote the regeneration and functional recovery after nerve injury becomes a focus.
Physical therapy is safe、non-invasion、convenient and effective ,which is favored by the clinical. A large number of studies suggest that pulsed electromagnetic fields can increase bone mineral density, and promote the fracture healing. It is often used in treating the osteoporosis. What’s more, some reports said that pulsed electromagnetic fields can promote the peripheral nerve injury repairing, however, some do not support. Therefore, it is necessary to do the related experimental researches to make its effectiveness and the parameters clearly, also the mechanism. Providing the evidences in treating the peripheral nerve injury to realize the clinical values.
Methods and materials:
Experiment 1 According to the principle of randomized, 30 SD rats were divided into five groups: the normal control (group A), model control (group B), 0.3mT 2Hz (group C), 0.3mT 10Hz (group D), 0.3mT 40Hz (group E), each 6. Except the normal control group, the other rats were clamped into sciatic nerve injury model according to the literature. After finishing the model, we gave them location. C、D、E group accepted the treatment , the PEMF’s field strength was 0.3mT, the frequency was 2Hz, 10Hz, 40Hz respectively. 2h/time, 2times/day, 30days altogether according the literature. the normal control group and model control group gave no intervention. Determined the sciatic nerve function index (SFI) and nerve conduction velocity(NCV) to all rats.
Experiment 2 Select 30 SD rats, divided into 3 groups: model control(group A)、normal control (group B)and treatment (group C), each 10. Except the normal control group, the other rats were received clamped sciatic nerve model according to the experiment 1. The treatment group were received 0.3mT 2Hz PEMF (according to the experiment 1’s result),2h/time, 2times/day, model control group and normal control group were not given any treatment. Except the normal control group,we took 5 rats from the other two groups respectively,Then the nerves which was 5mm away from the clamped were sampled at the 1~(st), 2~(nd), 4~(th) week respectively, observed by electron microscope and immunohistoc-hemistry. Image pattern analysis of axon was done at the 4th week after operation to model control group and treatment group.
Results:
In the same strength and time, 2Hz group’s SFI and NCV were higher than other frequency groups significantly (P<0.05). 2Hz PEMF make the course of Wallerian degeneration accelerated、schwann’s cell proliferation increased significantly、regeneration of axon improved obviously. The NGFof the treatment group expressed more than the model control group (P<0.05).
Conclusion:
0.3mT 2Hz PEMF may be suitable for treating the sciatic nerve injury. one of the mechanism may be: PEMF accelerated the course of Wallerian degeneration,Schwann’s cell proliferation,regeneration of axon, and the expression of NGF in nerve’s fiber.
引文
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[35]梅全喜,毕焕新,主编.现代中药药理手册.第1版,北京:中国中医药出版社,1998;324-585
[1] Liu Yinchun, You Huaming. Cell ultrastructure of ganoderma lacidum sprinkled with magnetized water [J]. Journal of Fujian college of Forestry, 2001,21(3):220-223.
[2] Liu Yinchun.Study on the law of experiment of wood absorbed 5.43um infrared and its penetrability for the infrared[J].Journal of Fujian College of Forestry,2002,22(3):227-231.
[3] Hao Bolin.Physics and biology[J].Physics, 2003, 32(6): 353-359.
[4] Xie Yuzhang.The Physics of Crystalline Liquid[M].Beijing:Technology Publisher, 1988.
[5] Komura Shigeyuki OU-YANG Zhongcan. High-and Low-pitch helicaI structures of tilted chiral lipid bilayers [J].Physical Review Letters,1998,81(2):473-476.
[6] Liebert L. Liquid Crystals[M].New York:Academic Press,1978:188-205.
[7] Luben RA,Cain CD, Chen MC-Y. et al.Effects of electromagnetic stimuling in bone and bone cells in vitro:inhibition of responses to parathyroid hormone by low energy, low frequency fields[J].Proc Nat Acad Sci,1982,79(3):4180.
[8] Li Rusheng. Nonbalance Energetics and DissipationStructure[M] Beijing:Tsinghua University Publisher,1986.
[9] Liu Changjun,Wang Baoyi,Zhang Hong,et al. A study on the diathesis function mechanism of cell model imposed by impulse electromagnetic field [J].Journalof Biomedical Engineering,1998,15(3):294-297.
[10]郭瑞华,黄噻清,贾鸿妹,等.脉冲电磁场促进周围神经再生的实验研究[J].中国康复医学杂志,1999,14(3):112-114.
[11]周田华,李主一,周中英,等.脉冲电磁场对周围神经再生的影响[J].中国临床康复,2003,7(2):208-209.
[12] Byers JM,Clark KF, Thompson GC.Effect of pulsed electromagnetic stimulationon facial nerve regeneration [J].Arch Otolaryngol Headneck Surg,1998,124 (4):383-389.
[13] Walker JL, et al. Enhancement of functional recovery following a crush lesion to the rat sciatic nerve by exposure to pulsed electromagnetic fields. Exp Neurol, 1994, 125(2): 302-305.
[14]刘永辉,侯希敏,李海霞,等.脉冲电刺激促进周围神经再生的应用[J].骨与关节损伤杂志,2000,12(1):342-343
[15] Mert T, et al. Regenerative effects of pulsed magnetic field on injured peripheral nerves. Altern Ther Health Med. 2006, 12(5): 42-49.
[16]周田华,李主一,周中英.等脉冲电磁场促进周围神经再生的实验观察.中国矫形外科杂志. 1985.5 160-161.
[17] Shah JP, Midkiff P, Brandt PC, et al. Growth and differentiation of PC6 cells: the effects of pulsed electromagnetic fields [J]. Bioelectromagnetics, 2001,22 (4): 267-271.
[18]房元章,余晓君,张红锋,等.低频脉冲电场对PC12细胞酪氨酸羟化酶和多巴胺合成的影响[J].细胞生物学杂志,2007,29(3):78-80.
[19]余晓君,张红锋,等.低频脉冲对PC12细胞突起生长和NGF受体分布的影响[J]细胞生物学杂志. 2004, 26:421-424.
[20] Longo FM, Yang T, Hamilton S. et al. Electromagnetic fields influence NGF acivity and levels following sciatic nerve transection [J]. J Neurosci Res, 1999, 55(2): 230-237.
[1] Dubovy P. Laminin molecules in freeze-treated nerve segments are associated with migrating Schwann cells that display the corresponding alpha6 betal integrin receptor[J]. Glia, 2001,33(1):36-44.
[2] Lnoue M, Hojo T, Yano T. et a1.The effects of electro acupuncture on peripheral nerve regeneration in rats[J]. Acupunct Med,2003,21(1-2):9-l7.
[3] Chen YS, Hu CL, Hsieh CL, et a1.Effects of percutaneous electrical stimulation on peripheral nerve regeneration using silicone rubber chambers [J].J Biomed Mater Res,2001, 57(4):541-549.
[4] Lazar DA, Curra FP, Molar B. et a1.Acceleration of recovery after injury to the peripheral nervous system using ultrasound and other therapeutic modalities[J].Neurosurg Clin N Am,2001,12(2):353-357.
[5] Poo MM. Motor reflex reponses elicited by cutaneous simulation in the regeneration nerve of man. [J]Annual review of biophysics’and bioengine 1981,10:245.
[6] Hinkle L, et al. The direction of growth of differentiating neurons and myoblasts inan applied electric fields. [J].physical 1981, 314:121.
[7] Patel N.B, Poo MM, Perturbation of neuritis growth by pulsed and focal electric fields [J] Neurosis 1984, 4:2 939.
[8] Bongens, RB, ETAL, Bioelectricity and regeneration initiation of frog limb regeneration by minute current [J] comp Neurd 1986,250:168.
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[12] Kolin A, Bril NO, Broberg PJ. Stimulation of irritable tissues by means of an alternating magnetic field. [J] Proc Soc Exp Biol.1953, 102(1):251-253
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[1] Liu Yinchun, You Huaming. Cell ultrastructure of ganoderma lacidum sprinkled with magnetized water [J]. Journal of Fujian college of Forestry, 2001,21(3):220-223.
[2] Liu Yinchun.Study on the law of experiment of wood absorbed 5.43um infrared and its penetrability for the infrared[J].Journal of Fujian College of Forestry,2002,22(3):227-231.
[3] Hao Bolin.Physics and biology[J].Physics, 2003, 32(6): 353-359.
[4] Xie Yuzhang.The Physics of Crystalline Liquid[M].Beijing:Technology Publisher, 1988.
[5] Komura Shigeyuki OU-YANG Zhongcan. High-and Low-pitch helicaI structures of tilted chiral lipid bilayers [J].Physical Review Letters,1998,81(2):473-476.
[6] Liebert L. Liquid Crystals[M].New York:Academic Press,1978:188-205.
[7] Luben RA,Cain CD, Chen MC-Y. et al.Effects of electromagnetic stimuling in bone and bone cells in vitro:inhibition of responses to parathyroid hormone by low energy, low frequency fields[J].Proc Nat Acad Sci,1982,79(3):4180.
[8] Li Rusheng. Nonbalance Energetics and DissipationStructure[M] Beijing:Tsinghua University Publisher,1986.
[9] Liu Changjun,Wang Baoyi,Zhang Hong,et al. A study on the diathesis function mechanism of cell model imposed by impulse electromagnetic field [J].Journalof Biomedical Engineering,1998,15(3):294-297.
[10]郭瑞华,黄噻清,贾鸿妹,等.脉冲电磁场促进周围神经再生的实验研究[J].中国康复医学杂志,1999,14(3):112-114.
[11]周田华,李主一,周中英,等.脉冲电磁场对周围神经再生的影响[J].中国临床康复,2003,7(2):208-209.
[12] Byers JM,Clark KF, Thompson GC.Effect of pulsed electromagnetic stimulationon facial nerve regeneration [J].Arch Otolaryngol Headneck Surg,1998,124 (4):383-389.
[13] Walker JL, et al. Enhancement of functional recovery following a crush lesion to the rat sciatic nerve by exposure to pulsed electromagnetic fields. Exp Neurol, 1994, 125(2): 302-305.
[14]刘永辉,侯希敏,李海霞,等.脉冲电刺激促进周围神经再生的应用[J].骨与关节损伤杂志,2000,12(1):342-343
[15] Mert T, et al. Regenerative effects of pulsed magnetic field on injured peripheral nerves. Altern Ther Health Med. 2006, 12(5): 42-49.
[16]周田华,李主一,周中英.等脉冲电磁场促进周围神经再生的实验观察.中国矫形外科杂志. 1985.5 160-161.
[17] Shah JP, Midkiff P, Brandt PC, et al. Growth and differentiation of PC6 cells: the effects of pulsed electromagnetic fields [J]. Bioelectromagnetics, 2001,22 (4): 267-271.
[18]房元章,余晓君,张红锋,等.低频脉冲电场对PC12细胞酪氨酸羟化酶和多巴胺合成的影响[J].细胞生物学杂志,2007,29(3):78-80.
[19]余晓君,张红锋,等.低频脉冲对PC12细胞突起生长和NGF受体分布的影响[J]细胞生物学杂志. 2004, 26:421-424.
[20] Longo FM, Yang T, Hamilton S. et al. Electromagnetic fields influence NGF acivity and levels following sciatic nerve transection [J]. J Neurosci Res, 1999, 55(2): 230-237.
[1] Dubovy P. Laminin molecules in freeze-treated nerve segments are associated with migrating Schwann cells that display the corresponding alpha6 betal integrin receptor[J]. Glia, 2001,33(1):36-44.
[2] Lnoue M, Hojo T, Yano T. et a1.The effects of electro acupuncture on peripheral nerve regeneration in rats[J]. Acupunct Med,2003,21(1-2):9-l7.
[3] Chen YS, Hu CL, Hsieh CL, et a1.Effects of percutaneous electrical stimulation on peripheral nerve regeneration using silicone rubber chambers [J].J Biomed Mater Res,2001, 57(4):541-549.
[4] Lazar DA, Curra FP, Molar B. et a1.Acceleration of recovery after injury to the peripheral nervous system using ultrasound and other therapeutic modalities[J].Neurosurg Clin N Am,2001,12(2):353-357.
[5] Poo MM. Motor reflex reponses elicited by cutaneous simulation in the regeneration nerve of man. [J]Annual review of biophysics’and bioengine 1981,10:245.
[6] Hinkle L, et al. The direction of growth of differentiating neurons and myoblasts inan applied electric fields. [J].physical 1981, 314:121.
[7] Patel N.B, Poo MM, Perturbation of neuritis growth by pulsed and focal electric fields [J] Neurosis 1984, 4:2 939.
[8] Bongens, RB, ETAL, Bioelectricity and regeneration initiation of frog limb regeneration by minute current [J] comp Neurd 1986,250:168.
[9] Zanakis MF. The use of DC electric fields to promote regeneration in the mamarian nerve system. [J]Acup Electro: Res, 1990, 15:185.
[10] Beveridge JA, et al. Faciliared regeneration in the rat peripheral nerve system using appied Electric fields.[J] Plast Reconstr surg, 1988, 83:301.
[11] Kern JM. Electrical simulation of nerve regeneration in the rat, the early effects evaluated By a vibrating probe and electron microscopy.[J] Neuroscience, 1991, 40:93.
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