推拿对坐骨神经损伤大鼠生长相关蛋白表达影响的研究
摘要
[目的]
基于推拿治疗坐骨神经损伤具有良好的临床疗效,从实验角度探寻坐骨神经损伤经推拿治疗后,机体在感觉功能及运动功能方面的改善效应,并通过对背根神经节中生长相关蛋白表达变化的研究,进一步揭示推拿治疗坐骨神经损伤的起效机理及作用途径。
[方法]
以SD大鼠为实验动物,制备坐骨神经夹持损伤模型,选取患侧殷门穴、承山穴、阳陵泉穴,每天用按摩手法模拟仪于以上三穴施以揉法、点法和拨法,通过对其斜板试验和足底部光热耐痛阈的检测,从行为学角度探寻坐骨神经损伤经推拿治疗后肢体运动功能和感觉功能的恢复情况；通过观察背根神经节脊髓、坐骨神经以及腓肠肌HE染色的变化,从形态学角度探寻推拿对神经元再生修复的促进效应；通过检测背根神经节及腓肠肌中生长相关蛋白表达的变化,探寻推拿促进损伤神经再生修复的机制,揭示推拿治疗坐骨神经损伤的起效机理和作用途径。
[结果]
1.行为学
造模7天后,模型组大鼠的斜板试验结果下降,与正常组比较具有统计学意义。推拿治疗10天后,推拿治疗组斜板试验结果高于模型组、模型对照组,趋于正常。推拿治疗可以使坐骨神经损伤大鼠腓肠肌肌力明显改善,促进其运动功能的恢复。
造模7天后,模型组大鼠右侧足底部光热耐痛阈升高,与正常组比较有统计学意义。推拿治疗10天后,推拿治疗组大鼠右侧足底部光热耐痛阈明显低于模型组,高于正常组,与模型组有统计学意义。推拿治疗可以提高坐骨神经损伤大鼠的痛觉敏感程度,促进其感觉功能的恢复。
2.病理学
正常情况下,背根神经节HE染色可见其神经元多数呈圆形,胞质呈嗜酸性,体积较大；胞核呈嗜碱性,染色浅,大而圆；核仁清楚,胞体外周有卫星细胞包绕；脊髓HE染色可见脊髓腹角细胞胞核位于细胞中央,尼氏体均匀分布。
通过对模型组的观察,DRG内部分神经元出现中央染色质溶解的现象,神经元有不同程度的坏死,细胞出现肿胀、变形和溶解的现象,胞浆的染色不均匀,细胞的形态也大小不一；脊髓HE染色显示细胞出现肿胀、胞核移向边缘尼氏体溶解消失等变性现象。说明坐骨神经损伤后,会导致背根神经节和脊髓腹角运动神经元胞体死亡。
通过对推拿治疗组的观察,DRG内神经元形状渐规则,大多为圆形,细胞肿胀及变形的现象不明显；脊髓腹角神经元部分细胞核移向边缘,部分细胞核位于中央,尼氏体溶解,细胞排列在完整性、方向性等方面好于模型组和模型对照组。说明推拿对坐骨神经损伤后感觉神经元和运动神经元的修复具有一定的促进作用。
3.生长相关蛋白
正常组及假手术组大鼠GAP-43在背根神经节及腓肠肌中有少量的表达,阳性的胞浆呈浅棕褐色,胞核呈淡蓝色；神经损伤7天后,模型组及模型对照组大鼠GAP-43在背根神经节及腓肠肌中的表达情况较正常组染色加深,免疫组化平均光密度增高,存在差异；推拿治疗10天后,模型组及模型对照组大鼠GAP-43在背根神经节及腓肠肌中的免疫组化平均光密度较正常组有一定程度增高,推拿治疗组大鼠GAP-43在背根神经节及腓肠肌中免疫组化平均光密度显著增高,染色程度和范围明显优于模型组,表达情况更为明显,较其余四组均存在差异。说明推拿治疗可以促进GAP-43在背根神经节及腓肠肌中的表达。
[结论]
推拿可以提高坐骨神经损伤大鼠的痛觉敏感程度,提高患侧腓肠肌的肌力及肌肉恢复率,从而促进坐骨神经损伤大鼠运动功能和感觉功能的恢复；其作用途径为对背根神经节感觉神经元及脊髓腹角运动神经元的营养和保护；起效机理之一为促进背根神经节及腓肠肌中生长相关蛋白的表达以促进损伤神经的再生修复,为损伤神经提供良好的微环境。
【Objective】
Based on better clinical effect of massage therapy on sciatic nerve injury, this study is to research the recovery on both sensory and motor function after massage therapy. Meanwhile, through the research on changes of GAP-43expression in DRG, the study can explore massage's approach and central mechanism on the treatment of sciatic nerve injury.
【Methods】
Taking SD rats as experimental animals to build clamping injured model, the study used massage simulator on UB37, BL57and GB34. And also, solar-thermal pain threshold resistance and inclined plate test is used from a behavioral point of view to explore the recovery on sensory and motor function after therapy. Through observing the variation of DRG, the spinal marrow, sciatic nerve and the HE of gastrocnemius, the promotion of therapy's effect on neuronal regeneration is sought on aspect of morphology. Through the research on changes of GAP-43expression in DRG and gastrocnemius muscles, approach and mechanism is revealed by massage therapy on sciatic nerve injury.
【Results】
1. Ethology
Seven days after made the model, results of inclined plate group declined which has statistical significance compared with normal group. Ten days after massage therapy, curative effect of inclined plate group is better than sham operation group, model group and control group. It means that massage therapy can improve gastrocnemius muscle for the sciatic nerve injury and promote the recovery of motor function.
Seven days after made the model, solar-thermal pain threshold resistance in model group is increased and it has statistical significance compared with normal group. Ten days after massage therapy, solar-thermal pain threshold resistance in therapy group is obviously lower than model group and higher than normal group. It has statistical significance compared with model group. It showed massage therapy can improve pain sensitivity for the sciatic nerve injury and promote the recovery of sensory function.
2. Pathology
Under normal circumstances, the neuron of DRG HE stain is visibly round and cytoplasm is eosinophilic and the cell volume is a bit larger. Cell nucleus is basophilic, staining light, large and round. Cell nucleolus is clear with satellite cells enveloping. HE in the spinal marrow is visible and cell nucleus is located in the central cells. Nissl body distributed uniformly of the cell bodies.
After sciatic nerve injured, part of DRG neurons appeared that chromatin dissolved. Neurons have different degrees of necrosis. Some cells were swelled and dissolved with some deformation. Cytoplasm staining wasn't even with form size differ. HE staining of the spinal cord cell swelled with degeneration phenomenon. The nucleus moved toward the edge and nissl body dissolved and disappeared. It showed that after sciatic nerve has injured, DRG and motor neuron cell of spinal cord ventral-horn died.
After massage therapy, DRG neuron is in the shape of gradual rules, mostly round. Cell is swelled and deformation phenomenon is not obvious. Part of ventral-horn nerve nucleus moved toward the edge and part of the nucleus is located in the central. Nissl body dissolved, cells array is better than model group and model control group in integrity and direction. Massage therapy can promote the recovery of both sensory and motor neuron after the sciatic nerve injured.
3. Growth associated protein
GAP-43in normal group and sham operation group has some expression in DRG and gastrocnemius muscles. Positive neurons cytoplasm appeared light brown, nucleus appeared pale blue. After seven days of sciatic nerve injured, GAP-43expression from model group and model control group in DRG and gastrocnemius muscles is deepen staining than normal group. AOD was increased, where there are differences. After ten days of massage therapy, AOD of GAP-43from model group and model control group in DRG and gastrocnemius muscles has a bit increase than normal group. AOD of GAP-43in DRG and AOD of GAP-43in gastrocnemius from massage therapy group significantly increased, staining extent and scope significantly better than model group. The expression is more obvious than the other four groups which has statistical significance.
[Conclusion]
Massage therapy can promote pain sensitivity of sciatic nerve injured rats, it also can increase gastrocnemius muscle's recovery rate. Both sensory and motor function is enhanced on the sciatic nerve injury. It effected on sensory neurons in DRG and motor neurons in the spinal marrow. The mechanism is for the promotion of GAP-43expression in both DRG and gastrocnemius muscles'growth-related proteins to promote the regeneration of injured nerves, and provide a good microcnvironment for the injured nerve.
基于推拿治疗坐骨神经损伤具有良好的临床疗效,从实验角度探寻坐骨神经损伤经推拿治疗后,机体在感觉功能及运动功能方面的改善效应,并通过对背根神经节中生长相关蛋白表达变化的研究,进一步揭示推拿治疗坐骨神经损伤的起效机理及作用途径。
[方法]
以SD大鼠为实验动物,制备坐骨神经夹持损伤模型,选取患侧殷门穴、承山穴、阳陵泉穴,每天用按摩手法模拟仪于以上三穴施以揉法、点法和拨法,通过对其斜板试验和足底部光热耐痛阈的检测,从行为学角度探寻坐骨神经损伤经推拿治疗后肢体运动功能和感觉功能的恢复情况；通过观察背根神经节脊髓、坐骨神经以及腓肠肌HE染色的变化,从形态学角度探寻推拿对神经元再生修复的促进效应；通过检测背根神经节及腓肠肌中生长相关蛋白表达的变化,探寻推拿促进损伤神经再生修复的机制,揭示推拿治疗坐骨神经损伤的起效机理和作用途径。
[结果]
1.行为学
造模7天后,模型组大鼠的斜板试验结果下降,与正常组比较具有统计学意义。推拿治疗10天后,推拿治疗组斜板试验结果高于模型组、模型对照组,趋于正常。推拿治疗可以使坐骨神经损伤大鼠腓肠肌肌力明显改善,促进其运动功能的恢复。
造模7天后,模型组大鼠右侧足底部光热耐痛阈升高,与正常组比较有统计学意义。推拿治疗10天后,推拿治疗组大鼠右侧足底部光热耐痛阈明显低于模型组,高于正常组,与模型组有统计学意义。推拿治疗可以提高坐骨神经损伤大鼠的痛觉敏感程度,促进其感觉功能的恢复。
2.病理学
正常情况下,背根神经节HE染色可见其神经元多数呈圆形,胞质呈嗜酸性,体积较大；胞核呈嗜碱性,染色浅,大而圆；核仁清楚,胞体外周有卫星细胞包绕；脊髓HE染色可见脊髓腹角细胞胞核位于细胞中央,尼氏体均匀分布。
通过对模型组的观察,DRG内部分神经元出现中央染色质溶解的现象,神经元有不同程度的坏死,细胞出现肿胀、变形和溶解的现象,胞浆的染色不均匀,细胞的形态也大小不一；脊髓HE染色显示细胞出现肿胀、胞核移向边缘尼氏体溶解消失等变性现象。说明坐骨神经损伤后,会导致背根神经节和脊髓腹角运动神经元胞体死亡。
通过对推拿治疗组的观察,DRG内神经元形状渐规则,大多为圆形,细胞肿胀及变形的现象不明显；脊髓腹角神经元部分细胞核移向边缘,部分细胞核位于中央,尼氏体溶解,细胞排列在完整性、方向性等方面好于模型组和模型对照组。说明推拿对坐骨神经损伤后感觉神经元和运动神经元的修复具有一定的促进作用。
3.生长相关蛋白
正常组及假手术组大鼠GAP-43在背根神经节及腓肠肌中有少量的表达,阳性的胞浆呈浅棕褐色,胞核呈淡蓝色；神经损伤7天后,模型组及模型对照组大鼠GAP-43在背根神经节及腓肠肌中的表达情况较正常组染色加深,免疫组化平均光密度增高,存在差异；推拿治疗10天后,模型组及模型对照组大鼠GAP-43在背根神经节及腓肠肌中的免疫组化平均光密度较正常组有一定程度增高,推拿治疗组大鼠GAP-43在背根神经节及腓肠肌中免疫组化平均光密度显著增高,染色程度和范围明显优于模型组,表达情况更为明显,较其余四组均存在差异。说明推拿治疗可以促进GAP-43在背根神经节及腓肠肌中的表达。
[结论]
推拿可以提高坐骨神经损伤大鼠的痛觉敏感程度,提高患侧腓肠肌的肌力及肌肉恢复率,从而促进坐骨神经损伤大鼠运动功能和感觉功能的恢复；其作用途径为对背根神经节感觉神经元及脊髓腹角运动神经元的营养和保护；起效机理之一为促进背根神经节及腓肠肌中生长相关蛋白的表达以促进损伤神经的再生修复,为损伤神经提供良好的微环境。
【Objective】
Based on better clinical effect of massage therapy on sciatic nerve injury, this study is to research the recovery on both sensory and motor function after massage therapy. Meanwhile, through the research on changes of GAP-43expression in DRG, the study can explore massage's approach and central mechanism on the treatment of sciatic nerve injury.
【Methods】
Taking SD rats as experimental animals to build clamping injured model, the study used massage simulator on UB37, BL57and GB34. And also, solar-thermal pain threshold resistance and inclined plate test is used from a behavioral point of view to explore the recovery on sensory and motor function after therapy. Through observing the variation of DRG, the spinal marrow, sciatic nerve and the HE of gastrocnemius, the promotion of therapy's effect on neuronal regeneration is sought on aspect of morphology. Through the research on changes of GAP-43expression in DRG and gastrocnemius muscles, approach and mechanism is revealed by massage therapy on sciatic nerve injury.
【Results】
1. Ethology
Seven days after made the model, results of inclined plate group declined which has statistical significance compared with normal group. Ten days after massage therapy, curative effect of inclined plate group is better than sham operation group, model group and control group. It means that massage therapy can improve gastrocnemius muscle for the sciatic nerve injury and promote the recovery of motor function.
Seven days after made the model, solar-thermal pain threshold resistance in model group is increased and it has statistical significance compared with normal group. Ten days after massage therapy, solar-thermal pain threshold resistance in therapy group is obviously lower than model group and higher than normal group. It has statistical significance compared with model group. It showed massage therapy can improve pain sensitivity for the sciatic nerve injury and promote the recovery of sensory function.
2. Pathology
Under normal circumstances, the neuron of DRG HE stain is visibly round and cytoplasm is eosinophilic and the cell volume is a bit larger. Cell nucleus is basophilic, staining light, large and round. Cell nucleolus is clear with satellite cells enveloping. HE in the spinal marrow is visible and cell nucleus is located in the central cells. Nissl body distributed uniformly of the cell bodies.
After sciatic nerve injured, part of DRG neurons appeared that chromatin dissolved. Neurons have different degrees of necrosis. Some cells were swelled and dissolved with some deformation. Cytoplasm staining wasn't even with form size differ. HE staining of the spinal cord cell swelled with degeneration phenomenon. The nucleus moved toward the edge and nissl body dissolved and disappeared. It showed that after sciatic nerve has injured, DRG and motor neuron cell of spinal cord ventral-horn died.
After massage therapy, DRG neuron is in the shape of gradual rules, mostly round. Cell is swelled and deformation phenomenon is not obvious. Part of ventral-horn nerve nucleus moved toward the edge and part of the nucleus is located in the central. Nissl body dissolved, cells array is better than model group and model control group in integrity and direction. Massage therapy can promote the recovery of both sensory and motor neuron after the sciatic nerve injured.
3. Growth associated protein
GAP-43in normal group and sham operation group has some expression in DRG and gastrocnemius muscles. Positive neurons cytoplasm appeared light brown, nucleus appeared pale blue. After seven days of sciatic nerve injured, GAP-43expression from model group and model control group in DRG and gastrocnemius muscles is deepen staining than normal group. AOD was increased, where there are differences. After ten days of massage therapy, AOD of GAP-43from model group and model control group in DRG and gastrocnemius muscles has a bit increase than normal group. AOD of GAP-43in DRG and AOD of GAP-43in gastrocnemius from massage therapy group significantly increased, staining extent and scope significantly better than model group. The expression is more obvious than the other four groups which has statistical significance.
[Conclusion]
Massage therapy can promote pain sensitivity of sciatic nerve injured rats, it also can increase gastrocnemius muscle's recovery rate. Both sensory and motor function is enhanced on the sciatic nerve injury. It effected on sensory neurons in DRG and motor neurons in the spinal marrow. The mechanism is for the promotion of GAP-43expression in both DRG and gastrocnemius muscles'growth-related proteins to promote the regeneration of injured nerves, and provide a good microcnvironment for the injured nerve.
引文
[1]薛立功.经筋系统的生理结构与解剖内涵[A].中国针灸学会经筋诊治专业委员会.中国针灸学会经筋诊治专业委员会成立大会论文集(2009年)重点论文选编[C].中国针灸学会经筋诊治专业委员会：,2009：77-89
[2]王战朝.名老中医李金明诊疗周围神经损伤经验[J].中国中医骨伤科杂志,2000,(01)：64-65.
[3]廉武星.筋痹辨治浅析[J].河南中医.1999,19(3)：38
[4]李元明.身痛逐瘀汤加减治疗腰椎间盘突出症[J].中国中医骨伤科杂志,2001,4(9)：39-40.
[5]徐枫.桃红饮加味治疗痰疲痹阻型痹症32例[J].四川中医,1999,17(1)：26-27
[6]罗跃东.补肾安络汤治疗坐骨神经痛60例[J].陕西中医,2004,25(12)：1097-1098.
[7]苏绍华.麻黄附子细辛汤加味治疗坐骨神经痛60例[J].江西中医药,2008,39(308)：53.
[8]柳哲,三乌坐痛饮治疗坐骨神经痛80例[J].陕西中医,2008,9(29)：1190-1191
[9]郭团茂,刘森,张银刚.独活寄生汤加减治疗椎间盘源性下腰痛67例[J].中医正骨,2010,22(10)：73-74
[10]陈宝强,陈霞,田凤琴.独活寄生汤加味治疗中老年坐骨神经痛88例临床观察[J].甘肃中医,2003,16(11)：25-26
[11]仇丽伟,杨建丰.通痹汤治疗原发性坐骨神经痛30例[J].河南中医.2002,22(4)：35
[12]陈选惠.独活寄生汤合补阳还五汤化裁治疗坐骨神经痛60例[J].四川中医,2005,1(23)：38.
[13]雷学礼.加味三痹汤治疗痹症125例[J].陕西中医,2009,30(8)：1011.
[14]韩卢丽,张江涛.补阳还五汤治疗下肢神经损伤56例[J].四川中医,2003,7(21)：79-80.
[15]熊卫红,张葆现.愈痹汤配合穴位注射治疗坐骨神经痛66例[J].陕西中医2009,30(8)：1012.
[16]王喜芳.龙胆泻肝汤加减治疗非根性坐骨神经痛200例疗效观察[J].甘肃中医,2007,20(8)：36
[17]罗发生.清热利湿化瘀法治疗坐骨神经痛63例[J].江西中医药.2004,7(35)：45
[18]高瑞琴.针刺环跳、委中穴为主治疗坐骨神经痛168例[J].河北中医,2002, 1(24)：41.
[19]刘淑印.针刺中药并用治疗坐骨神经痛57例[J].实用中医内科杂志,2008,22(8)：65.
[20]尹国红,王怀兰.针刺治疗梨状肌综合征46例[J].光明中医,2009,12(24)：2312
[21]尹根起,陈伟江,张志芬,等.针刺治疗坐骨神经痛135例临床报告[J].针灸临床杂志,2003,19(5)：19
[22]盛生宽,盛全成.齐刺法为主治疗坐骨神经痛38例临床观察[J].国医论坛,2006,4(21)：27.
[23]韦迪.齐刺法加温针灸治疗干性坐骨神经痛60例[J].国际医药卫生导报,2003,9(7)：75-76
[24]雷国华,安军明.齐刺法治疗梨状肌综合征60例疗效观察[J].现代中医药,2005,3：51.
[25]陈美仁,王萍,成钢,等.苍龟探穴针法治疗坐骨神经痛临床观察.中华中医药学刊,2007,25(12)：2546-2547
[26]刘娜,邓玉霞.苍龟探穴治疗梨状肌综合征疗效观察[J].上海针灸杂志,2007,26(11)：25-26
[27]艾双春,蒲一宏,周祖刚,等.针灸飞经走气法为主对地震伤员周围神经损伤的康复[J].医学信息,2011,2：752-753.
[28]王泽显.经穴电针治疗老年根性坐骨神经痛临床观察[J].中国针灸,2009,29(2)：126-128
[29]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[J].中国实用神经疾病杂志,2008,8(11)：35-36.
[30]肖华强.推拿并电针治疗医源性坐骨神经损伤1例[J].中国中医急症,2003,12(5)：482.
[31]曹友德,王敏华,王敏,等.电针夹脊穴治疗根性坐骨神经痛临床观察[J].上海针灸杂志,2003,22(10)：19-20
[32]张磊.针刀、针灸治疗慢性腰腿痛166例临床疗效观察[J],内蒙古中医药,010,29(13)：86.
[33]张伟民.针刀治疗梨状肌综合征的临床观察[J].山西医药杂志,2009,38(4)：374-375
[34]单赤军.小针刀治疗梨状肌综合征128例疗效观察[J].武警医学院学报,2009,18(9)：796
[35]左同军.针刀治疗干性坐骨神经痛78例[J].现代中西医结合杂志,2009, 18(8)：897-89
[36]于天源.按摩推拿学[M].北京：中国协和医科大学出版社,2009.
[37]乔欣军,手法治疗急性臀上皮神经损伤38例报告[J].中国中医骨伤科杂志,2003,3(11)：40-41.
[38]杨国芳.按摩治疗坐骨神经痛[J].按摩与导引,2007,23(12)：39-40
[39]陈于阳.侧扳法为主治疗腰椎间盘突出症108例[J].中国中医急症,2010,19(3)： 514-515
[40]李岩,周震,贺小靖,徐卫革.火针疗法治疗痹证的临床疗效研究[J].针灸临床杂志,2003,(04)：31-34+63.
[41]尹晶,陈邦国,王俊华,王刚,李陈盈.温针灸治疗腰椎间盘突出症的临床观察[J].湖北中医杂志,2008,(12)：44-46.
[42]冯凯,马建国,李继安,徐明,周云涛,吕宪民,高海凤.活血康元汤对Wistar大鼠坐骨神经损伤保护作用的实验研究[J].中国煤炭工业医学杂志,2000,(08)：859-861.
[43]石杜鹃；凌丽；薛金伟；补阳还五汤药浴对大鼠周围神经损伤再生影响的实验研究[J].时珍国医国药,2008,(05)：1066-1067.
[44]钱叶斌,陶有略,黄翠芬,武一曼.补阳还五汤促进周围神经损伤后神经元存活的实验研究[J].实用手外科杂志,2002,(03)：152-154.
[45]薛金伟；姜志刚；韩冬；朱清远；尹维田.补阳还五汤药浴对大鼠周围神经再生组织形态学的影响[J].中国老年学杂志,2008,(06)：523-524.
[46]魏光如,姜保国,赵富强,张宏波复方红芪减方对周围神经再生影响的实验研究[J].中华手外科杂志,2004,(03)：51-53.
[47]杨德梅；阿丽娅；魏光如；董建强；姜保国；复方红芪减方提取液对坐骨神经损伤后表达bFGF、 NGF和Trk的影响[J].中华手外科杂志,2006,(05)：308-310.
[48]尹宗生,顾玉东,顾映红,朱腾方中药治疗对大鼠坐骨神经损伤后神经生长因子蛋白表达的影响[J].中华手外科杂志,2003,(01)：57-59.
[49]满毅；徐广辉；林浩东；复方神肌再生冲剂对大鼠坐骨神经损伤后生长相关蛋白-43表达的影响[J].上海中医药杂志,2011,(04)：63-66.
[50]吕丹；朱雷；李建鑫；复方丹参注射液和甲基泼尼松龙联合治疗周围神经损伤的实验研究[J].华南国防医学杂志,2009,(04)：25-26+37.
[51]邵水金,单宝枝,余安胜.电针对大鼠坐骨神经损伤后轴突转运的影响.针剌研究,1998,23(1)：66
[52]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
[53]郭义,石田寅夫,李庆雯,等.不同频率电针对周围神经再生与修复影响的临床与实验研究[J].天津中医药大学学报,2006,25(3)：177-181.
[54]李庆雯,石田寅夫,郭义.不同频率电针对大鼠坐骨神经损伤后神经组织形态学与骨骼肌肌电图的影响[J].中国针灸,2005,25(3)：217-210
[55]张晓晖.电针治疗对坐骨神经损伤大鼠NGF、 CNTF、 BDNF、 bFGF表达的影响[D].导师：于天源.：北京中医药大学,2011
[56]李澎涛,陶之理.电针、NGF对臂丛前索损伤再生过程中脊髓运动神经元GAP-43表达的影响[J].神经解剖学杂志,2000,16(1)：67-69.
[57]贾立,余学东,卢公标,黄纯聪,袁世全,.139例不同原因周围神经损伤临床分析[J].重庆医学,2007,(18).
[58]刘勇,康志学,穆广态,等.周围神经损伤120例的疗效分析[J].宁夏医学杂志,2007,29(7)：634-635
[59]邵长敏,韩建梅,孙华婧.48例儿童臀部坐骨神经注射性损伤治疗临床探讨.中国全科医学,2004,7(5)：352.
[60]郝兴伟,郝坚明,李德光,.周围神经损伤的临床分析[J].中国医药指南,2010,(3).
[61]吴克坚,殷代昌,陈家臻.周围神经损伤的实验与临床观察[J].临床军医杂志,2001,(1).
[62]孙占胜,邱春晖,赵孔波.游离神经移植修复周围神经缺损100例[J].山东医药,2003,43(27)：21-22
[63]张建众,李维宾,包小义,徐吉,蔡国平.接枝水凝胶硅胶膜修复周围神经损伤缺损的初步临床应用[J].中国临床康复,2002,(6).
[64]陈晓东.周围神经损伤与修复[J].创伤外科杂志,2004,6(6)：478-450.
[65]姚朝娅,丁新生,王杏英.人神经生长因子治疗坐骨神经干性损伤的疗效观察[J].南京医科大学学报,1999, 19(1)：45-46
[66]梁伦高,梁崇礼,翁龙江,杨举伦,李云玫,夏正武bFGF对晚期周围神经再生作用的电生理评价[J].昆明医学院学报,2001,(04)：21-23.
[67]史福东,刘东,李长江.神经生长因子在周围神经损伤中的应用[J].现代中西医结合杂志,2008,17(27)：4281-4282.
[68]顾立强,陈国奋,相大勇,朱立军,裴国献.免疫抑制剂FK506加速周围神经损伤修复后功能恢复的初步临床报告[J].中国创伤骨科杂志,2001,(3).
[69]陈华规,董方红,刘芸,等.腰椎旁神经阻滞治疗椎管外软组织损害引起的坐骨神经痛.长江大学学报,2006,3(2)：257-259
[70]陈伟.用肌电图仪治疗臂丛及周围神经损伤的临床应用[D].大连医科大学大连医科大学,2006.
[71]董红让,米永,李澜,.高压氧配合显微外科技术治疗高原军训中周围神经损伤的临床效果观察[J].解放军医学杂志,2009,(2).
[72]田德虎,张英泽,米立新,赵峰,韩久卉,张继春,韩金豹.分米波在周围神经损伤后s-100蛋白表达变化中作用的实验研究[J].中国康复医学杂志,2004,(04)：29-31.
[73]刘大庸,胡耀民,钟世镇.周围神经系统的微环境.中国临床解剖学杂志1991,9：244.
[74]罗永湘,方火皇,庞清江,等.神经生长因子对周围神经运动纤维再生的影响[J].中华手外科杂志,1997, 13(1)：3-5.
[75]Santos FX, Bilbao G, Rodrigo J， et al. Experimental model for local administration of nerve growth factor in microsurgical reconnection[J] Microsurgery,1995,16(2):71-76.
[76]Burnett MG, Zager EL. Pathophysiology of peripheral nerve injury:a briei review. Neurosurg Focus,2005,16:El.
[77]Stirling, V. The dance of the growth cones-where to next? Trends Neurosci, 1995; 18:111-115
[78]Alex, L.K.Growth cones and the cues that repel them. Trends Neurosci.1996; 19:507-513
[79]Benowitz,-L-I:Routtenberg,-A..GAP-43:an intrinsic determinant of neuronal development and plasticity[J]. Trends Neurosci,1997, Feb:20(2):84-91.
[80]Tolner EA, van Vliet EA, Holtmaat AJ, et al. GAP-43 mRNA and protein expression in the hippocampal and parahippocampal region during the course of epileptogenesis in rats[J]. Eur J Neurosci,2003,17(11):2369-2380.
[81]Nacimiento W et al:J Neurobiol 22:413; 1993
[82]Hall SMJ Neurosci Res 21:820,1992
[83]Golding JP and Tonge DA:Neuroscience52:415,1993
[84]Liabotis,-S; Schreyer,-D-J. Magnitude of GAP-43 intuction following peripheral axotomy of adult rat dorsal root ganglion neurons is independent of lesion distance.Exp-Neurol.1995 Sep; 135(1):28-35
[85]张在强；曹世俭；王拥军；大鼠坐骨神经损伤后背根神经节再生微环境变化[A].第九次全国神经病学学术大会论文汇编[C].中国广东广州：,2006
[86]刘光久,李振强,殷玉芹,宋林,孙建森.神经损伤与再生中神经元GAP-43mRNA表达的原位杂交研究[J].解剖学报,2002,(06)：659-661.
[87]Benowitz,-L-I:Routtenberg,-A..GAP-43:an intrinsic determinant of neuronal development and plastieity[J].Trends Neurosei,1997, Feb:20(2):84-91.
[88]Chakravarthy B, Rashid A, Brown L, et al.Association of GAP-43 (neuromodulin) with microtubule-associated protein MAP-2 in neuronal cells[J]. Biochemical and Biophysical Research Communications,2008, 371(4):679-683.
[89]宋晔；推拿牵引治疗腰椎间盘突出症65例[J].中医药临床杂志,2011,(11)：978.
[90]叶玉霞；葛平丽；机牵配合膀胱经推拿治疗腰椎间盘突出症[J].中国医学工程,2011,(10)：154+158.
[91]郝双阶；田朝晖；自拟推拿手法治疗腰椎间盘突出症120例[J].湖北民族学院学报(医学版),2010,(01)：53-54.
[92]王炳；推拿手法及复位治疗腰椎间盘突出症临床疗效观察[J].按摩与导引,2007,(08)：22-24.
[93]李军；推拿治疗梨状肌损伤综合征32例[J].河南科技大学学报(医学版),2006,(04)：272-273.
[94]丛德毓；邓伟；李新建；推拿手法治疗梨状肌综合征96例临床观察[J].中国社区医师(综合版),2007,(12)：87.
[95]沈海军；王瑞强；李德君；手法推拿治疗梨状肌综合征33例[J].中国社区医师(医学专业),2012,(01)：216.
[96]杨杰.推拿治疗梨状肌综合征100例[J].现代康复,2000,(13)：124.
[97]刘志诚.推拿牵引镇痛调整作用与5-羟色胺等含量变化的研究[J].中西医结合杂志,1985,5(10)：615-617.
[98]张建华,孙安达.推拿牵引治疗腰椎间盘突出症的疗效观察及血中单胺类物质含量变化分析[J].中国骨伤,1999,12(5)：47-48.
[99]陈全珠,刘冰怀,焦洁,等.推拿牵引镇痛调整唾液中5-羟色胺的变化.疼痛学杂志,1995；3(4)：149-150.
[100]李静.推拿治疗腰椎间盘突出症的临床及其机理研究[D].导师：王道全.：山东中医药大学,2003.
[101]韩东伟.推拿对坐骨神经损伤大鼠行为学和形态学的影响[D].导师：于天源.：北京中医药大学,2008.
[102]吴凡.推拿对坐骨神经损伤大鼠胶质细胞源性神经营养因子的影响[D].导师：于天源.：北京中医药大学,2010.
[103]韦景斌.推拿干预对周围神经损伤大鼠脊髓背角中NTFs表达变化的研究[D].导师：于天源.：北京中医药大学,2011.
[104]袁之琳.推拿对大鼠坐骨神经损伤后NTFs的影响[D].导师：于天源.：北京中医药大学,2011.
[105]Hasegawa.K..A new method of measuring functional recovery after crushing the peripheral nerves in unanesthetized and unrestrained rats.Experientia, 1978; 34:272
[106]Paul DS and John DH. pli-tubulin and GAP-43mRNA expression ir chronically injured neurons of the red nucleus after a second spinal corc injury[J].Experimental Neurology[J],2003, Vol.183 (2):537-547
[107]丰岩清,郭云良,石秉霞.脑缺血再灌注后生长相关蛋白和突触素表达及其意义.青岛大学医学院学报,2000,12.15,36(4)：247
[2]王战朝.名老中医李金明诊疗周围神经损伤经验[J].中国中医骨伤科杂志,2000,(01)：64-65.
[3]廉武星.筋痹辨治浅析[J].河南中医.1999,19(3)：38
[4]李元明.身痛逐瘀汤加减治疗腰椎间盘突出症[J].中国中医骨伤科杂志,2001,4(9)：39-40.
[5]徐枫.桃红饮加味治疗痰疲痹阻型痹症32例[J].四川中医,1999,17(1)：26-27
[6]罗跃东.补肾安络汤治疗坐骨神经痛60例[J].陕西中医,2004,25(12)：1097-1098.
[7]苏绍华.麻黄附子细辛汤加味治疗坐骨神经痛60例[J].江西中医药,2008,39(308)：53.
[8]柳哲,三乌坐痛饮治疗坐骨神经痛80例[J].陕西中医,2008,9(29)：1190-1191
[9]郭团茂,刘森,张银刚.独活寄生汤加减治疗椎间盘源性下腰痛67例[J].中医正骨,2010,22(10)：73-74
[10]陈宝强,陈霞,田凤琴.独活寄生汤加味治疗中老年坐骨神经痛88例临床观察[J].甘肃中医,2003,16(11)：25-26
[11]仇丽伟,杨建丰.通痹汤治疗原发性坐骨神经痛30例[J].河南中医.2002,22(4)：35
[12]陈选惠.独活寄生汤合补阳还五汤化裁治疗坐骨神经痛60例[J].四川中医,2005,1(23)：38.
[13]雷学礼.加味三痹汤治疗痹症125例[J].陕西中医,2009,30(8)：1011.
[14]韩卢丽,张江涛.补阳还五汤治疗下肢神经损伤56例[J].四川中医,2003,7(21)：79-80.
[15]熊卫红,张葆现.愈痹汤配合穴位注射治疗坐骨神经痛66例[J].陕西中医2009,30(8)：1012.
[16]王喜芳.龙胆泻肝汤加减治疗非根性坐骨神经痛200例疗效观察[J].甘肃中医,2007,20(8)：36
[17]罗发生.清热利湿化瘀法治疗坐骨神经痛63例[J].江西中医药.2004,7(35)：45
[18]高瑞琴.针刺环跳、委中穴为主治疗坐骨神经痛168例[J].河北中医,2002, 1(24)：41.
[19]刘淑印.针刺中药并用治疗坐骨神经痛57例[J].实用中医内科杂志,2008,22(8)：65.
[20]尹国红,王怀兰.针刺治疗梨状肌综合征46例[J].光明中医,2009,12(24)：2312
[21]尹根起,陈伟江,张志芬,等.针刺治疗坐骨神经痛135例临床报告[J].针灸临床杂志,2003,19(5)：19
[22]盛生宽,盛全成.齐刺法为主治疗坐骨神经痛38例临床观察[J].国医论坛,2006,4(21)：27.
[23]韦迪.齐刺法加温针灸治疗干性坐骨神经痛60例[J].国际医药卫生导报,2003,9(7)：75-76
[24]雷国华,安军明.齐刺法治疗梨状肌综合征60例疗效观察[J].现代中医药,2005,3：51.
[25]陈美仁,王萍,成钢,等.苍龟探穴针法治疗坐骨神经痛临床观察.中华中医药学刊,2007,25(12)：2546-2547
[26]刘娜,邓玉霞.苍龟探穴治疗梨状肌综合征疗效观察[J].上海针灸杂志,2007,26(11)：25-26
[27]艾双春,蒲一宏,周祖刚,等.针灸飞经走气法为主对地震伤员周围神经损伤的康复[J].医学信息,2011,2：752-753.
[28]王泽显.经穴电针治疗老年根性坐骨神经痛临床观察[J].中国针灸,2009,29(2)：126-128
[29]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[J].中国实用神经疾病杂志,2008,8(11)：35-36.
[30]肖华强.推拿并电针治疗医源性坐骨神经损伤1例[J].中国中医急症,2003,12(5)：482.
[31]曹友德,王敏华,王敏,等.电针夹脊穴治疗根性坐骨神经痛临床观察[J].上海针灸杂志,2003,22(10)：19-20
[32]张磊.针刀、针灸治疗慢性腰腿痛166例临床疗效观察[J],内蒙古中医药,010,29(13)：86.
[33]张伟民.针刀治疗梨状肌综合征的临床观察[J].山西医药杂志,2009,38(4)：374-375
[34]单赤军.小针刀治疗梨状肌综合征128例疗效观察[J].武警医学院学报,2009,18(9)：796
[35]左同军.针刀治疗干性坐骨神经痛78例[J].现代中西医结合杂志,2009, 18(8)：897-89
[36]于天源.按摩推拿学[M].北京：中国协和医科大学出版社,2009.
[37]乔欣军,手法治疗急性臀上皮神经损伤38例报告[J].中国中医骨伤科杂志,2003,3(11)：40-41.
[38]杨国芳.按摩治疗坐骨神经痛[J].按摩与导引,2007,23(12)：39-40
[39]陈于阳.侧扳法为主治疗腰椎间盘突出症108例[J].中国中医急症,2010,19(3)： 514-515
[40]李岩,周震,贺小靖,徐卫革.火针疗法治疗痹证的临床疗效研究[J].针灸临床杂志,2003,(04)：31-34+63.
[41]尹晶,陈邦国,王俊华,王刚,李陈盈.温针灸治疗腰椎间盘突出症的临床观察[J].湖北中医杂志,2008,(12)：44-46.
[42]冯凯,马建国,李继安,徐明,周云涛,吕宪民,高海凤.活血康元汤对Wistar大鼠坐骨神经损伤保护作用的实验研究[J].中国煤炭工业医学杂志,2000,(08)：859-861.
[43]石杜鹃；凌丽；薛金伟；补阳还五汤药浴对大鼠周围神经损伤再生影响的实验研究[J].时珍国医国药,2008,(05)：1066-1067.
[44]钱叶斌,陶有略,黄翠芬,武一曼.补阳还五汤促进周围神经损伤后神经元存活的实验研究[J].实用手外科杂志,2002,(03)：152-154.
[45]薛金伟；姜志刚；韩冬；朱清远；尹维田.补阳还五汤药浴对大鼠周围神经再生组织形态学的影响[J].中国老年学杂志,2008,(06)：523-524.
[46]魏光如,姜保国,赵富强,张宏波复方红芪减方对周围神经再生影响的实验研究[J].中华手外科杂志,2004,(03)：51-53.
[47]杨德梅；阿丽娅；魏光如；董建强；姜保国；复方红芪减方提取液对坐骨神经损伤后表达bFGF、 NGF和Trk的影响[J].中华手外科杂志,2006,(05)：308-310.
[48]尹宗生,顾玉东,顾映红,朱腾方中药治疗对大鼠坐骨神经损伤后神经生长因子蛋白表达的影响[J].中华手外科杂志,2003,(01)：57-59.
[49]满毅；徐广辉；林浩东；复方神肌再生冲剂对大鼠坐骨神经损伤后生长相关蛋白-43表达的影响[J].上海中医药杂志,2011,(04)：63-66.
[50]吕丹；朱雷；李建鑫；复方丹参注射液和甲基泼尼松龙联合治疗周围神经损伤的实验研究[J].华南国防医学杂志,2009,(04)：25-26+37.
[51]邵水金,单宝枝,余安胜.电针对大鼠坐骨神经损伤后轴突转运的影响.针剌研究,1998,23(1)：66
[52]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
[53]郭义,石田寅夫,李庆雯,等.不同频率电针对周围神经再生与修复影响的临床与实验研究[J].天津中医药大学学报,2006,25(3)：177-181.
[54]李庆雯,石田寅夫,郭义.不同频率电针对大鼠坐骨神经损伤后神经组织形态学与骨骼肌肌电图的影响[J].中国针灸,2005,25(3)：217-210
[55]张晓晖.电针治疗对坐骨神经损伤大鼠NGF、 CNTF、 BDNF、 bFGF表达的影响[D].导师：于天源.：北京中医药大学,2011
[56]李澎涛,陶之理.电针、NGF对臂丛前索损伤再生过程中脊髓运动神经元GAP-43表达的影响[J].神经解剖学杂志,2000,16(1)：67-69.
[57]贾立,余学东,卢公标,黄纯聪,袁世全,.139例不同原因周围神经损伤临床分析[J].重庆医学,2007,(18).
[58]刘勇,康志学,穆广态,等.周围神经损伤120例的疗效分析[J].宁夏医学杂志,2007,29(7)：634-635
[59]邵长敏,韩建梅,孙华婧.48例儿童臀部坐骨神经注射性损伤治疗临床探讨.中国全科医学,2004,7(5)：352.
[60]郝兴伟,郝坚明,李德光,.周围神经损伤的临床分析[J].中国医药指南,2010,(3).
[61]吴克坚,殷代昌,陈家臻.周围神经损伤的实验与临床观察[J].临床军医杂志,2001,(1).
[62]孙占胜,邱春晖,赵孔波.游离神经移植修复周围神经缺损100例[J].山东医药,2003,43(27)：21-22
[63]张建众,李维宾,包小义,徐吉,蔡国平.接枝水凝胶硅胶膜修复周围神经损伤缺损的初步临床应用[J].中国临床康复,2002,(6).
[64]陈晓东.周围神经损伤与修复[J].创伤外科杂志,2004,6(6)：478-450.
[65]姚朝娅,丁新生,王杏英.人神经生长因子治疗坐骨神经干性损伤的疗效观察[J].南京医科大学学报,1999, 19(1)：45-46
[66]梁伦高,梁崇礼,翁龙江,杨举伦,李云玫,夏正武bFGF对晚期周围神经再生作用的电生理评价[J].昆明医学院学报,2001,(04)：21-23.
[67]史福东,刘东,李长江.神经生长因子在周围神经损伤中的应用[J].现代中西医结合杂志,2008,17(27)：4281-4282.
[68]顾立强,陈国奋,相大勇,朱立军,裴国献.免疫抑制剂FK506加速周围神经损伤修复后功能恢复的初步临床报告[J].中国创伤骨科杂志,2001,(3).
[69]陈华规,董方红,刘芸,等.腰椎旁神经阻滞治疗椎管外软组织损害引起的坐骨神经痛.长江大学学报,2006,3(2)：257-259
[70]陈伟.用肌电图仪治疗臂丛及周围神经损伤的临床应用[D].大连医科大学大连医科大学,2006.
[71]董红让,米永,李澜,.高压氧配合显微外科技术治疗高原军训中周围神经损伤的临床效果观察[J].解放军医学杂志,2009,(2).
[72]田德虎,张英泽,米立新,赵峰,韩久卉,张继春,韩金豹.分米波在周围神经损伤后s-100蛋白表达变化中作用的实验研究[J].中国康复医学杂志,2004,(04)：29-31.
[73]刘大庸,胡耀民,钟世镇.周围神经系统的微环境.中国临床解剖学杂志1991,9：244.
[74]罗永湘,方火皇,庞清江,等.神经生长因子对周围神经运动纤维再生的影响[J].中华手外科杂志,1997, 13(1)：3-5.
[75]Santos FX, Bilbao G, Rodrigo J， et al. Experimental model for local administration of nerve growth factor in microsurgical reconnection[J] Microsurgery,1995,16(2):71-76.
[76]Burnett MG, Zager EL. Pathophysiology of peripheral nerve injury:a briei review. Neurosurg Focus,2005,16:El.
[77]Stirling, V. The dance of the growth cones-where to next? Trends Neurosci, 1995; 18:111-115
[78]Alex, L.K.Growth cones and the cues that repel them. Trends Neurosci.1996; 19:507-513
[79]Benowitz,-L-I:Routtenberg,-A..GAP-43:an intrinsic determinant of neuronal development and plasticity[J]. Trends Neurosci,1997, Feb:20(2):84-91.
[80]Tolner EA, van Vliet EA, Holtmaat AJ, et al. GAP-43 mRNA and protein expression in the hippocampal and parahippocampal region during the course of epileptogenesis in rats[J]. Eur J Neurosci,2003,17(11):2369-2380.
[81]Nacimiento W et al:J Neurobiol 22:413; 1993
[82]Hall SMJ Neurosci Res 21:820,1992
[83]Golding JP and Tonge DA:Neuroscience52:415,1993
[84]Liabotis,-S; Schreyer,-D-J. Magnitude of GAP-43 intuction following peripheral axotomy of adult rat dorsal root ganglion neurons is independent of lesion distance.Exp-Neurol.1995 Sep; 135(1):28-35
[85]张在强；曹世俭；王拥军；大鼠坐骨神经损伤后背根神经节再生微环境变化[A].第九次全国神经病学学术大会论文汇编[C].中国广东广州：,2006
[86]刘光久,李振强,殷玉芹,宋林,孙建森.神经损伤与再生中神经元GAP-43mRNA表达的原位杂交研究[J].解剖学报,2002,(06)：659-661.
[87]Benowitz,-L-I:Routtenberg,-A..GAP-43:an intrinsic determinant of neuronal development and plastieity[J].Trends Neurosei,1997, Feb:20(2):84-91.
[88]Chakravarthy B, Rashid A, Brown L, et al.Association of GAP-43 (neuromodulin) with microtubule-associated protein MAP-2 in neuronal cells[J]. Biochemical and Biophysical Research Communications,2008, 371(4):679-683.
[89]宋晔；推拿牵引治疗腰椎间盘突出症65例[J].中医药临床杂志,2011,(11)：978.
[90]叶玉霞；葛平丽；机牵配合膀胱经推拿治疗腰椎间盘突出症[J].中国医学工程,2011,(10)：154+158.
[91]郝双阶；田朝晖；自拟推拿手法治疗腰椎间盘突出症120例[J].湖北民族学院学报(医学版),2010,(01)：53-54.
[92]王炳；推拿手法及复位治疗腰椎间盘突出症临床疗效观察[J].按摩与导引,2007,(08)：22-24.
[93]李军；推拿治疗梨状肌损伤综合征32例[J].河南科技大学学报(医学版),2006,(04)：272-273.
[94]丛德毓；邓伟；李新建；推拿手法治疗梨状肌综合征96例临床观察[J].中国社区医师(综合版),2007,(12)：87.
[95]沈海军；王瑞强；李德君；手法推拿治疗梨状肌综合征33例[J].中国社区医师(医学专业),2012,(01)：216.
[96]杨杰.推拿治疗梨状肌综合征100例[J].现代康复,2000,(13)：124.
[97]刘志诚.推拿牵引镇痛调整作用与5-羟色胺等含量变化的研究[J].中西医结合杂志,1985,5(10)：615-617.
[98]张建华,孙安达.推拿牵引治疗腰椎间盘突出症的疗效观察及血中单胺类物质含量变化分析[J].中国骨伤,1999,12(5)：47-48.
[99]陈全珠,刘冰怀,焦洁,等.推拿牵引镇痛调整唾液中5-羟色胺的变化.疼痛学杂志,1995；3(4)：149-150.
[100]李静.推拿治疗腰椎间盘突出症的临床及其机理研究[D].导师：王道全.：山东中医药大学,2003.
[101]韩东伟.推拿对坐骨神经损伤大鼠行为学和形态学的影响[D].导师：于天源.：北京中医药大学,2008.
[102]吴凡.推拿对坐骨神经损伤大鼠胶质细胞源性神经营养因子的影响[D].导师：于天源.：北京中医药大学,2010.
[103]韦景斌.推拿干预对周围神经损伤大鼠脊髓背角中NTFs表达变化的研究[D].导师：于天源.：北京中医药大学,2011.
[104]袁之琳.推拿对大鼠坐骨神经损伤后NTFs的影响[D].导师：于天源.：北京中医药大学,2011.
[105]Hasegawa.K..A new method of measuring functional recovery after crushing the peripheral nerves in unanesthetized and unrestrained rats.Experientia, 1978; 34:272
[106]Paul DS and John DH. pli-tubulin and GAP-43mRNA expression ir chronically injured neurons of the red nucleus after a second spinal corc injury[J].Experimental Neurology[J],2003, Vol.183 (2):537-547
[107]丰岩清,郭云良,石秉霞.脑缺血再灌注后生长相关蛋白和突触素表达及其意义.青岛大学医学院学报,2000,12.15,36(4)：247