摘要
目的：近年来研究表明,神经病理性疼痛的产生与脊髓背角神经元中枢敏感化,以及脊髓背角神经元突触可塑性长时程增强的产生有密切的关系。研究长时程增强,以及针刺是否能抑制长时程增强的产生,对于研究神经病理性疼痛以及针刺治疗神经病理性疼痛的机制都有非常重要的意义。本实验采用坐骨神经分支选择性损伤(SNI)大鼠模型,观察电针对神经病理性疼痛大鼠痛阈和脊髓背角突触传递长时程增强的抑制作用,以进一步探讨神经病理性疼痛的发病机制以及针刺镇痛的机理。
方法：健康成年雄性SD大鼠经筛选后,符合条件的大鼠随机分为假手术对照组和造模组。制备大鼠坐骨神经分支选择性损伤型神经病理性疼痛(SNI)模型。造模成功后随机将模型大鼠分为模型组和模型加电针干预组进行实验,模型组观察14日,于SNI术前、术后次日、第7天、第14天测定大鼠的机械痛阈；模型加电针干预组大鼠机械痛阈观察同模型组,但在第7天开始,用HANS穴位神经刺激仪2Hz,1mA电针环跳、委中穴30min,每日1次,连续7天。观察各组大鼠强直刺激后C纤维诱发场电位的变化,另外观察电针对模型组大鼠已形成并维持的LTP的抑制作用,即在模型组3h观察结束后,给予电针干预,参数时间同前,电针后观察1h。观察电针环跳、委中穴,对SNI模型大鼠机械痛阈和脊髓背角神经元突触传递LTP的干预作用。
结果：
1.SNI术后,大鼠双侧机械痛阈显著进行性降低,以术侧更为明显,与同时段假手术组比较有显著差异性(P<0.01)同时大鼠出现术侧后肢足趾卷曲、足外翻、以及步态异常等疼痛行为学的变化。
2.电针干预后,大鼠术侧的机械痛阈值由电针前的20.6±2.6g提高至38.5±4.7g,明显高于同时段模型组(P<0.01),同时其疼痛行为异常也明显减轻。
3.模型组大鼠C-纤维诱发场电位变化率在HFS诱导前、诱导后,以及诱导3h时分别为69.9±30.8%、224.9±98.5%和174.6±38.3%,,HFS诱导后C-纤维诱发场电位变化率显著高于基础平均值(P<0.01),而同时段假手术组HFS诱导后3h各个时段比较无统计学意义。
4.模型加电针干预组大鼠电针后,C-纤维诱发场电位变化率在HFS诱导前、诱导后,以及诱导3h时分别为75.6±33.1%、73.3±20.3%和62.9±20.8%,与假手术组相当,与模型组比较有显著性差异(P<0.01)。
5.对模型组已形成的脊髓背角神经元LTP,并稳定维持3 h的SNI模型大鼠施行电针干预,C-纤维诱发场电位变化率从电针前的182.8±29.7%降至34.0±16.3%,与电针前比较具有显著差异(P<0.01)。
结论：神经病理性疼痛的产生与脊髓背角神经元突触可塑性长时程增强的形成以及脊髓背角神经元的中枢敏化有着密切的关系。电针可以显著抑制C纤维诱发电位LTP的形成,电针对LTP的维持具有有显著的阻断作用。同时电针可显著改善神经病理性疼痛模型大鼠的疼痛行为学变化。提示电针可能是通过抑制脊髓背角神经元突触可塑性变化的形成,减弱脊髓背角神经元的敏感化而发挥镇痛作用。
Objective:In recent years, studies have shown that the generation of neuropathic pain is closely related to the and spinal cord dorsal horn neurons in sensitization,spinal cord dorsal horn neurons in long-term potentiation of synaptic plasticity.It has important significance to study acupuncture on treatment of neuropathic pain and neuropathic pain mechanism,long-term potentiation,and whether acupuncture can inhibit the production of long-term potentiation. This study is sciatic nerve injury in rat model of selective branches to observe the power for the pain threshold of neuropathic pain in rats and spinal cord dorsal horn synaptic transmission and long-term potentiation of the inhibitory effect in order to further explore the mechanism of acupuncture analgesia.
Methods:Healthy adult male SD rats after screening, eligible rats were randomly divided into sham operation group, model group, and electro acupuncture intervention group. Selective Preparation of sciatic nerve branch injury neuropathic pain model (SNI) model. The model group observed 14 days and observed SNI before surgery, the next day, the 7th day, the first 14 days of determination of the mechanical pain threshold in rats; model of electro-acupuncture intervention mechanical pain threshold in rats to observe the same model group, but in the first 7 day one, with the HANS acupoint nerve stimulator 2Hz,1mA electro-acupuncture stimulate Weizhong and Huantiao for 30min, once a day for 7 days. Another observation is the inhibition effect of LTP electro acupuncture for the model group has been formed and maintained by, that is to say,observed the model group after the end of 3h given electro-acupuncture interventions, the parameter of time with the former, electro-acupuncture observed after 1h. Observation intervention effect of LTP of synaptic transmission use electro-acupuncture Huantiao and Weizhong to the SNI rats with mechanical pain threshold and spinal cord dorsal horn neurons.
Results:1. After the operation of SNI, rats with bilateral mechanical pain threshold decreased significantly and more effect in operative side. At the same time, segment in sham group There were significant differences (P<0.01) while rats exhibited hind toe curling operative side, foot valgus, as well as pain and abnormal gait behavior changes.
2. After use electro-acupuncture,the mechanical side surgery pain threshold by electro-acupuncture before 20.6±2.6g increased to 38.5±4.7g, significantly higher than the same time segment model group (P<0.01), while significantly reducing their pain and abnormal behavior.
3. Model group C-fiber-evoked field potentials the rate of change in the HFS before induction, and after 3h induction, respectively for 69.9±30.8%,224.9±98.5% and 174.6±38.3%,, HFS induced C-fiber evoked field potential significantly higher than the rate of change of the basis of the average (P<0.01), while at the same segment in sham operation group at all time 3h after the induction of HFS was no statistical significance.
4. Model of electro-acupuncture plus intervention in rats after electro-acupuncture, C-fiber-evoked field potentials the rate of change in the HFS before induction, and after 3h induction, respectively for 75.6±33.1%,73.3±20.3% and 62.9±20.8%, and the sham-operated group equivalent compared with the model group were significantly different (P<0.01).
5. Group has been formed on the model of spinal cord dorsal horn neurons LTP, and to stabilize the maintenance of the SNI model rats 3 h the purposes of electro-acupuncture interventions, C-fiber-evoked field potentials from the electro-acupuncture before the rate of change of 182.8±29.7% down to 34.0±16.3%, with electro-acupuncture earlier significant differences (P<0.01).
Conclusion:The production of neuropathic pain and spinal cord dorsal horn neurons in synaptic plasticity long-term potentiation, as well as the formation of the spinal cord dorsal horn neurons of the central sensitization has close relationship. Electro-acupuncture can significantly inhibit C fiber-evoked potentials of the formation of LTP, electricity for the maintenance of LTP has a significant role in blocking. At the same time, electro-acupuncture can significantly improve the rat model of neuropathic pain, pain behavior change. Electro-acupuncture may be prompted by inhibiting the spinal cord dorsal horn neurons in synaptic plasticity changes in the formation of reduced spinal cord dorsal horn neurons in sensitization exert analgesic effect.
引文
[1]Bennett GJ.Neuropathic pain:new insights,new interventions [J].HospPract,1998,33:95-114.
[2]李琦,曾炳芳,王金武.神经病理性疼痛研究进展[J].中国疼痛医学杂志,2007,13,(4)：244-246.
[3]赵序利,赵松云,王珺楠.慢性疼痛的治疗[J].中国临床医生,2005,33(9)：6-8.
[4]DevorM.Neuropathic pain and injured nerve:peripheral mechanisms[J].BrMed Bull,1991,47(3):619-630.
[5]Yasuhiko K, Zhen ZX, Xiaoying W, et al. Distinct roles of Inntrix metalloprotcascs in the Early and late-phase development of neuropathic pain[J]. Nature Medicine,2008(14):331—336.
[6]Rollin MG, Mardini I. Advances in Neuropathic Pain[C]. American Pain Society(APS)26th Annum Scientific Meeting,2007.
[7]Scholz J, Broom DC, Youn DH, et al. Blocking caspase activity prevents transsynaptic neuronal apoptosis and the loss of inhibition in lamina II of the dorsal horn after peripheral nerve injury. [J].Neurosci, 2005,25:7317-7323.
[8]HUGHES DI SCOTT DT,TODD AJ et al,Lack of evidence for sprouting of Abeta afferents into the superficial laminas of the spinal cord dorsal horn after nerve section [J].Neurosci 2003,23(29):9491-9499
[9]Woolf CJ.Evidence for a central component of post-injury pain hypersensitivity[J].Nature,1983,306 (5944):686-688.
[10]Vikman KS,Duggan AW,Siddall PJ.Increased ability to induce long-term potentiation of spinal dorsal horn neurons in monoarthriticrats[J].Brain Research,2003,990(1-2):51—57.
[11]张红梅,周利君,刘先国.急性神经损伤引起脊髓背角C-纤维诱发电位长时程增强[J].生理学报,2004,56(5)：591-596.
[12]邢国刚,刘风雨,姚磊等.神经病理痛大鼠脊髓背角突触传递的长时程可塑性变化[J].北京大学学报(医学版),2003,3：226.
[13]Klein T, Magerl W, Hopf HC,et al Perceptual correlates of nociceptive long-term potentiation and long-term depression in humans[J].J Neurosci,2004,24:964-971.
[14]Soderling TR, Chang B,Bricky D Cellular signaling through multifunctional Ca2+/calmodulin-dependent protein kinase II. [J]. J Biol Chem,2001;267:3719-4204.
[15]Fang L, Wu J, Lin Q, Willis WD. Calcium-calmodulin dependent protein kinase II contributes to spinal cord central sensitization. [J] Neurosci,2002;22:4196-4204.
[16]Roberson ED,Sweat JD. Transient activation of cyclic AMP-dependent protein kinase during hippocam long-termpotentiaiton[J].Biol. Chem,1996,271 (48):30436-30441.
[17]杨红卫,胡晓东,刘先国等.CaMK II抑制剂阻抑脊髓背角C纤维诱发电位LTP的诱导和早期维持[J].中国神经科学杂志,2003 19(6)：374-378.
[18]张红梅,胡晓东,刘先国等.PKC在成年大鼠脊髓背角C-纤维诱发电位长时程增强的诱导和维持中的作用[J].中国病理生理杂志,2005,21(2)：252-255.
[19]杨红卫,张红梅,胡晓东等.PKA参与脊髓背角C-纤维诱发电位LTP的诱导和早期维持[J].中国病理生理杂志,2004,20(4)：513-517.
[20]De-Felici M,Pesce M. Growth factors in mouse primordial germ cell migration and proliferation[J].Prog Growth Factor Res,1994,5(2):135.
[21]Gjerstad J,Tiolsen A,Hole K.Induction of long-term potentiation of single wide dynamic rang neurones in the dorsal horn is inhibited by descending pathways[J].Pain,2001,91(3):263—268.
[22]Miletica G,Draganicb P,Pankratzc MT,et al.Muscimol prevents long-lasting potentiation of dorsal horn field potentials in rats with chronic constriction injury exhibiting decreased levels of t he GABAt ransporter GAT-1[J].Pain,2003,105:347—353.
[23]Tegeder I,Turco DD,Schmidtko A.et al.Reduced inflammatory hyperalgesia with preservation of acutet hermal nociception in mice lacking cGMP-dependent protein kinase I[J].PNAS,2004,101:3253-3257.
[24]张健,茹立强,李之望.电针对三叉神经痛大鼠痛行为反应的影响[J].陕西中医,2004,25(7)：660-662.
[25]李娜,李为民,陈颖波等.电针对完全弗氏佐剂性小鼠外周慢性炎症痛的缓解作用[J].中国针灸,2008,28(2)：122-125.
[26]梁繁荣,罗荣,刘雨星等.电针镇痛后效应与炎症局部5-HT、NE、DA含量关系的实验研究[J].中国中医基础医学杂志,2001,7(11)：52-55.
[27]张利泰,陈以国,成泽东.电针抗大鼠创伤痛免疫抑制效应的实验研究[J].辽宁中医杂志,2001.28(6)：379-380.
[28]李翠贤,闰丽萍,易建良,马骋。电针对神经病理性疼痛大鼠的干预作用及其脊髓EAAs含量的影响[J].上海针灸杂志,2008,27(1)：45-48.
[29]孙涛,宋文阁,姚尚龙等.电针降低神经病理性疼痛大鼠脊髓白细胞介素-6的表达[J].针刺研究,2006,37(5)：286-289.
[30]潘惠娟,王海燕,许建阳等.电针对实验性RA大鼠痛阈及脑干NO/NOS变化的研究[J].上海针灸杂志,2002,21(5)：48-50.
[31]姚凯,郭义,胡利民等.针刺镇痛与中枢神经系统神经元内外游离钙离子浓度的关系[J].天津中医药,2005,22(5)：399-400.
[32]郝卫方,张海,张志刚.头维穴重刺激为主治疗顽固性三叉神经痛32例[J].上海针灸,2009,28(2)：74.
[33]李伟洪.针刺治疗三叉神经痛58例[J].上海针灸杂志,2009,28(11)：665.
[34]苏静,程进.针刺治疗带状疱疹82例[J].山东中医杂志,2007,26(7)：486.
[35]王立群.针刺治疗带状疱疹78例[J].中医研究,2008,21(5)：245-246.
[36]王翠连.电针配合针刺治疗坐骨神经痛38例[J].山西职工医学院院学报,2008,18(3)：57.
[37]李晓帆,田佩庸.针刺治疗坐骨神经痛100例[J].吉林中医药,2004,24(11)：50.
[38]陈增辉,王立新.针灸治疗类风湿关节炎40例[J].长春中医药大学学报,2009,25(2)：247.
[39]钟伟泉.针刺加井穴放血治疗急性痛风性关节炎34例[J].实用医学杂志,2009,25(15)：2580-2581.
[40]Bennet GJ,Xie.Peripheral mononeuropathy in rat that produce disorders of pain sensation like those seen in man [J].Pain,1988;33:87-107.
[41]Seltzer Z. Novel behavioral model of neuropathic pain disorders produced in rats by partial sciatic nerve injury[J].Pain,1990;43:208-218.
[42]Kim SH.Chung JM.An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat [J].Pain,1992;50:355-363.
[43]Decosterd I,Woolf CJ.Spared nerve injury an animal model of persistent peripheral neuropathic pain[J].Pain,2000,87(2):149-158.
[44]余维豪,范维铭,蔡红等.“腰背委中求”的临床与机理研究[J].中国针灸,1997,(8)：503.
[45]杨向红,王彦青,高秀等,电针对大鼠神经痛痛敏分数的影响[J].针刺研究.2002,27(1)：60-63.
[46]朱兵.针灸的科学基础[M].青岛出版社,1998：281-283.
[47]张红梅,祁英杰,李永勇等.刺激Aδ纤维抑制背角c纤维诱发电位长时程增强[J].中国临床康复,2003,7(10)：1510-1512.
[48]Chen J, Sandkuhler J. Induction of homosynaptic long-term depression at spinal synapses of sensory delta-fibers requires activation of metabotropic glutamate receptors. Neuroscience[J].2000,98(1):141-148.
[49]马骋,李翠贤,易建良等.电针对大鼠神经病理性疼痛模型DRG和脊髓Glu和Asp含量的影响[J].针刺研究,2008,33(4)：250-254.
[50]马骋,冯克辉,闫丽萍.电针抑制iGluRs激动剂鞘内给药易化的大鼠脊髓背角突触传递LTP[J].中国疼痛医学杂志.,2005,15,(4)：236-239.
[51]李永新,梅镇彤.长时程增强的形成机制及其与学习记忆的相关性[J].生理科学进展,1993,24(3)：278-280.
[1]李永新,梅镇彤.长时程增强的形成机制及其与学习记忆的相关性[J].生理科学进展.,1993,24(3)：278-280
[2]Woolf CJ.Evidence for a central component of post-injury pain hypersensitivity[J].Nature,1983,306 (5944):686-688
[3]Vikman KS,Duggan AW,Siddall PJ.Increased ability to inducelong-term potentiation of spinal dorsal horn neurons in monoarthriticrats[J].Brain Research,2003,990(1-2):51—57.
[4]张红梅,周利君,刘先国.急性神经损伤引起脊髓背角C-纤维诱发电位长时程增强[J]生理学报,2004,56(5)：591-596
[5]邢国刚,刘风雨,姚磊等.神经病理痛大鼠脊髓背角突触传递的长时程可塑性变化[J].北京大学学报(医学版),2003,3：226.
[6]Klein T, Magerl W, Hopf HC,et al Perceptual correlates of nociceptive long-term potentiation and long-term depression in humans [J]. J Neurosci,2004,24:964—971.
[7]Soderling TR,Chang B,Bricky D Cellular signaling through multifunctional Ca2+/calmodulin-dependent protein kinase Ⅱ.[J] Biol Chem.2001;267:3719-4204.
[8]Fang L,Wu J,Lin Q,Willis WD.Calcium-calmodulin dependent protein kinase Ⅱ contributes to spinal cord central sensitization Neurosci 2002;22:4196-4204.
[9]Roberson ED, Sweat JD. Transient activation of cyclic AMP-dependent protein kinase during hippocam long-termpotentiaiton[J].J Biol.Chem,1996,271(48):30436-30441
[10]杨红卫,胡晓东,刘先国等.CaMK II抑制剂阻抑脊髓背角C纤维诱发电位LTP的诱导和早期维持[J].中国神经科学杂志,2003,19(6)：374-378.
[11]张红梅,胡晓东,刘先国等.PKC在成年大鼠脊髓背角C—纤维诱发电位长时程增强的诱导和维持中的作用[J].中国病理生理杂志,2005,21(2)：252-255
[12]杨红卫,张红梅,胡晓东等.PKA参与脊髓背角C—纤维诱发电位LTP的诱导和早期维持[J]中国病理生理杂志,2004,20(4)：513-517
[13]De-Felici M,Pesce M. Growth factors in mouse primordial germ cell migration and proliferation[J].Prog Growth Factor Res,1994,5(2):135
[14]Gjerstad J,Tiolsen A,Hole K.Induction of long-term potentiation of single wide dynamic rang neurones in the dorsal horn is inhibited by descending pathways[J].Pain,2001,91(3):263—268.
[15]Miletica G,Draganicb P, Pankratzc MT,et al.Muscimol prevents long-lasting potentiation of dorsal horn field potentials in rats with chronic constriction injury exhibiting decreased levels of t he GABAt ransporter GAT-1[J].Pain,2003,105:347—353.
[16]Tegeder I,Turco DD,Schmidtko A,et al.Reduced inflammatory hyperalgesia with preservation of acute hermal nociception in mice lacking cGMP-dependent protein kinase I[J].PNAS,2004,101:3253—3257.
[17]张健,茹立强,李之望.电针对三叉神经痛大鼠痛行为反应的影响[J].陕西中医,2004,25(7)：660-662.
[18]李娜,李为民,陈颖波等.电针对完全弗氏佐剂性小鼠外周慢性炎症痛的缓解作用[J].中国针灸,2008,28(2)：122-125.
[19]梁繁荣,罗荣,刘雨星等.[J].中国中医基础医学杂志,2001,7(11)：52-55.
[20]张利泰,陈以国,成泽东.电针抗大鼠创伤痛免疫抑制效应的实验研究[J].辽宁中医杂志,2001,28(6)：379-380.
[21]李翠贤,闰丽萍,马骋等.电针对神经病理性疼痛大鼠的干预作用及其脊髓EAAs含量的影响[J].上海针灸杂志,2008,27(1)：45-48.
[22]孙涛,宋文阁,姚尚龙等.电针降低神经病理性疼痛大鼠脊髓白细胞介素-6的表达[J].针刺研究,2006,37(5)：286-289.
[23]潘惠娟,王海燕,许建阳等.电针对实验性RA大鼠痛阂及脑干NO/NOS变化的研究[J].上海针灸杂志,2002,21(5)：48-50.
[24]姚凯,郭义,胡利民等.针刺镇痛与中枢神经系统神经元内外游离钙离子浓度的关系[J].天津中医药,2005,22(5)：399-400.
[25]朱兵.针灸的科学基础[M].青岛出版社1998：281-283.
[26]张红梅,祁英杰,李永勇等.刺激Aδ纤维抑制背角c纤维诱发电位长时程增强[J].中国临床康复,2003,7(10)：1510-1512.
[27]Chen J, Sandkuhler J.Induction of homosynaptic long-term depres-sion at spinal synapses of sensory a delta-fibers requires activation of metabotropic glutamate receptors. Neuroscience[J].2000,98(1):141-148.
[28]马骋,李翠贤,易建良等.电针对大鼠神经病理性疼痛模型DRG和脊髓Glu和Asp含量的影响[J].针刺研究,2008,33(4)：250-254.
[29]马骋,冯克辉,闫丽萍.电针抑制iGluRs激动剂鞘内给药易化的大鼠脊髓背角突触传递LTP[J].中国疼痛医学杂志,2005,15(4)：236-239.