内降钙素基因相关肽和μ阿片受体在腰椎间盘突出模型大鼠脊髓后角内的表达
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摘要
目的利用椎间孔内置管模拟椎间盘突出压迫脊神经根,观察模型鼠脊髓后角内降钙素基因相关肽(CGRP)和μ阿片受体(MOR)的表达,探讨腰椎间盘突出后致腰腿疼发生的可能机制。方法将18只SD大鼠随机分为模型组和对照组,模型组在大鼠椎间孔内置管,模拟腰椎间盘突出压迫脊神经根;对照组在大鼠椎间孔插管后旋即拔出。于术前、术后分别观察大鼠行为;采用足底力学感觉测量仪测定大鼠后肢机械痛阈,热板测痛仪测定大鼠左后足底的热刺激感受阈值;免疫荧光组织化学法观察CGRP和MOR在脊髓后角的形态学特点;Western blot检测CGRP和MOR在脊髓内的表达。结果模型组大鼠后肢运动受限,对照组于术后第7天基本恢复正常;术后第1、3和7天模型组的机械痛阈值和热痛阈值均小于同时间点对照组(P均<0.05);免疫荧光发现CGRP和MOR蛋白均表达于脊髓后角,且经Western blot检测发现CGRP含量模型组较对照组升高,而MOR蛋白含量模型组较对照组降低。结论腰椎间盘突出模型大鼠脊髓背角内CGRP和MOR蛋白表达改变,可能为腰椎间盘突出症腰腿痛的机制之一。
Objective To observe the expression of calcitonin gene-related peptide(CGRP) andμ opioid receptor(MOR) in the posterior horn of the spinal cord using the intervertebral foramen to simulate the intervertebral disc herniation,and then explores the possible mechanism of the occurrence of lumbar leg pain during lumbar disc herniation. Methods Eighteen SD rats were randomly divided into control group and model group. In the model group,a canal was inserted into the intervertebral foramen to simulate the compression of the spinal nerve root by lumbar disc herniation,in the control group,the cannula was inserted into the intervertebral foramen and then pulled out. Behavior of rats was observed before and after operation,mechanical pain threshold of hind limbs was measured by plantar mechanical sensory meter,thermal stimulation threshold of left hind plantar was measured by hot plate pain meter,morphological characteristics of CGRP and MOR in the posterior horn of spinal cord were observed by immunofluorescence histochemistry,CGRP and MOR expression in spinal cord were detected by Western blot. Results Compared with control group,the hind limb movement was restricted in the model group at day 7 after operation. Simultaneously,the threshold of mechanical pain and thermal pain in the model group was significantly lower than those of the control group at 1 d,3 d and 7 d after operation(P<0.05). The results of immunofluorescence showed that CGRP and MOR expressed in the posterior horn of spinal cord. According to Western blot,the expression of CGRP in the model group was higher than control group,while the content of MOR was lower than the control group.Conclusion The expression change of CGRP and MOR in spinal dorsal horn of rats with lumbar disc herniation is probably the mechanism of lumbar and leg pain in lumbar disc herniation.
Objective To observe the expression of calcitonin gene-related peptide(CGRP) andμ opioid receptor(MOR) in the posterior horn of the spinal cord using the intervertebral foramen to simulate the intervertebral disc herniation,and then explores the possible mechanism of the occurrence of lumbar leg pain during lumbar disc herniation. Methods Eighteen SD rats were randomly divided into control group and model group. In the model group,a canal was inserted into the intervertebral foramen to simulate the compression of the spinal nerve root by lumbar disc herniation,in the control group,the cannula was inserted into the intervertebral foramen and then pulled out. Behavior of rats was observed before and after operation,mechanical pain threshold of hind limbs was measured by plantar mechanical sensory meter,thermal stimulation threshold of left hind plantar was measured by hot plate pain meter,morphological characteristics of CGRP and MOR in the posterior horn of spinal cord were observed by immunofluorescence histochemistry,CGRP and MOR expression in spinal cord were detected by Western blot. Results Compared with control group,the hind limb movement was restricted in the model group at day 7 after operation. Simultaneously,the threshold of mechanical pain and thermal pain in the model group was significantly lower than those of the control group at 1 d,3 d and 7 d after operation(P<0.05). The results of immunofluorescence showed that CGRP and MOR expressed in the posterior horn of spinal cord. According to Western blot,the expression of CGRP in the model group was higher than control group,while the content of MOR was lower than the control group.Conclusion The expression change of CGRP and MOR in spinal dorsal horn of rats with lumbar disc herniation is probably the mechanism of lumbar and leg pain in lumbar disc herniation.
引文
[1]张朝佑.人体解剖学[M]. 3版.北京:人民卫生出版社,2009:2132.
[2]陈琼夏.优秀女子帆船470运动员腰椎间盘突出症特征分析[J].当代体育科技,2015(9):9-10.
[3]张钢林,孟庆林,吕钟毓.腰椎间盘突出症的综合治疗─附498例分析[J].北京体育大学学报,1997(1):50-53.
[4]朱章标,鹿红宁,张方同,等.中国优秀蹦床运动员运动损伤特征、原因及防治[J].中国体育科技,2014,50(1):43-47.
[5]李娟红,倪家骧.硬膜外腔植入髓核对大鼠脊髓背根神经节SP和CGRP表达的影响[J].颈腰痛杂志,2009,30(6):497-500.
[6] Yi JY,Yeon LJ,Chul-Kyu P. Resolvin E1 inhibits substance P-Induced potentiation of TRPV1 in primary sensory neurons[J].Mediators of Inflammation,2016,2016(11):1-9.
[7]邱新毓,鲁亚成,李金莲. CGRP样阳性终末在脊神经结扎模型大鼠脊髓背角浅层内的表达变化[J].神经解剖学杂志,2013,29(5):498-502.
[8]武俊芳,韩金珠,郭志坤,等.慢性神经痛大鼠脊髓背角P物质、降钙素基因相关肽的表达[J].郑州大学学报(医学版),2008,43(4):699-701.
[9] Krock E,Rosenzweig DH,ChabotdoréAJ,et al. Painful,degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors[J].Journal of Cellular&Molecular Medicine,2014,18(6):1213-1225.
[10]牛乐.内吗啡肽2参与镇痛效应的形态学和机能学研究[D].西安:第四军医大学,2012.
[11] Narita M,Ozaki S,Suzuki T. Endomorphin-induced motivational effect:differential mechanism of endomorphin-1 and endomorphin-2[J]. Japanese Journal of Pharmacology,2002,89(3):224.
[12]焦旭文,徐泽,李军平,等. EPO/NGF对大鼠背根节压迫性损伤的治疗作用[J].神经解剖学杂志,2016,32(3):293-300.
[13]焦旭文,王国平,李军平,等. EPO对大鼠腰神经根慢性压迫性损伤的保护作用[J].宁夏医科大学学报,2013,35(11):1199-1203.
[14]陈卫明,徐鸽,金若敏,等.腰椎间盘突出症病因机制的研究进展[J].时珍国医国药,2007,18(6):1502-1504.
[15]林世德,张俊江,臧宇杰,等.脊髓损伤相关中枢性疼痛与CGRP阳性的初级传入纤维再生的相关性研究[J].中国矫形外科杂志,2016,24(24):2274-2279.
[16] Kou ZZ,Wan FP,Bai Y,et al. Decreased endomorphin-2 andμ-Opioid receptor in the spinal cord are associated with painful diabetic neuropathy[J].Front Mol Neurosci,2016,9:80.
[17] Kumar A,Liu NJ,Madia PA,et al. Contribution of endogenous spinal endomorphin 2 to intrathecal opioid antinociception in rats is agonist dependent and sexually dimorphic[J]. J Pain,2015,16(11):1200-1210.
[18]李云庆.内吗啡肽在脊髓背角通过突触前机制发挥镇痛作用[C]//中华医学会疼痛学分会年会暨casp成立二十周年.2009.
[19] Sakurada S,Hayashi T,Yuhki M,et al. Differential antagonism of endomorphin-1 and endomorphin-2supraspinal antinociception by naloxonazine and 3-methylnaltrexone[J]. Peptides,2002,23(5):895-901.
[20] Delafoy L,Gelot A,Ardid D,et al. Interactive involvement of brain derived neurotrophic factor,nerve growth factor, and calcitonin gene related pep tide in colonic hypersensitivity in the rat[J].Gut,2006,55(7):940-945.
[21] Greenwell TN,Martin-schild S,Inglis FM,et al.Colocalization and shared distribution of endomorphins with substance P,calcitonin gene-related peptide,gamma-aminobutyric acid,and the mu opioid receptor[J]. J Comp Neurol,2007,503(2):319-333.
[22] Sanderson Nydahl K,Skinner K,Julius D,et al. Colocalization of endomorphin-2 and substance P in primary afferent nociceptors and effects of injury:a light and electron microscopic study in the rat[J].Eur J Neurosci,2004,19(7):1789-1799.
[23] Xie H,Woods JH,Traynor JR,et al. The spinal antinociceptive effects of endomorphins in rats:behavioral and G protein functional studies[J]. Anesth Analg,2008,106(6):1873-1881.
[2]陈琼夏.优秀女子帆船470运动员腰椎间盘突出症特征分析[J].当代体育科技,2015(9):9-10.
[3]张钢林,孟庆林,吕钟毓.腰椎间盘突出症的综合治疗─附498例分析[J].北京体育大学学报,1997(1):50-53.
[4]朱章标,鹿红宁,张方同,等.中国优秀蹦床运动员运动损伤特征、原因及防治[J].中国体育科技,2014,50(1):43-47.
[5]李娟红,倪家骧.硬膜外腔植入髓核对大鼠脊髓背根神经节SP和CGRP表达的影响[J].颈腰痛杂志,2009,30(6):497-500.
[6] Yi JY,Yeon LJ,Chul-Kyu P. Resolvin E1 inhibits substance P-Induced potentiation of TRPV1 in primary sensory neurons[J].Mediators of Inflammation,2016,2016(11):1-9.
[7]邱新毓,鲁亚成,李金莲. CGRP样阳性终末在脊神经结扎模型大鼠脊髓背角浅层内的表达变化[J].神经解剖学杂志,2013,29(5):498-502.
[8]武俊芳,韩金珠,郭志坤,等.慢性神经痛大鼠脊髓背角P物质、降钙素基因相关肽的表达[J].郑州大学学报(医学版),2008,43(4):699-701.
[9] Krock E,Rosenzweig DH,ChabotdoréAJ,et al. Painful,degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors[J].Journal of Cellular&Molecular Medicine,2014,18(6):1213-1225.
[10]牛乐.内吗啡肽2参与镇痛效应的形态学和机能学研究[D].西安:第四军医大学,2012.
[11] Narita M,Ozaki S,Suzuki T. Endomorphin-induced motivational effect:differential mechanism of endomorphin-1 and endomorphin-2[J]. Japanese Journal of Pharmacology,2002,89(3):224.
[12]焦旭文,徐泽,李军平,等. EPO/NGF对大鼠背根节压迫性损伤的治疗作用[J].神经解剖学杂志,2016,32(3):293-300.
[13]焦旭文,王国平,李军平,等. EPO对大鼠腰神经根慢性压迫性损伤的保护作用[J].宁夏医科大学学报,2013,35(11):1199-1203.
[14]陈卫明,徐鸽,金若敏,等.腰椎间盘突出症病因机制的研究进展[J].时珍国医国药,2007,18(6):1502-1504.
[15]林世德,张俊江,臧宇杰,等.脊髓损伤相关中枢性疼痛与CGRP阳性的初级传入纤维再生的相关性研究[J].中国矫形外科杂志,2016,24(24):2274-2279.
[16] Kou ZZ,Wan FP,Bai Y,et al. Decreased endomorphin-2 andμ-Opioid receptor in the spinal cord are associated with painful diabetic neuropathy[J].Front Mol Neurosci,2016,9:80.
[17] Kumar A,Liu NJ,Madia PA,et al. Contribution of endogenous spinal endomorphin 2 to intrathecal opioid antinociception in rats is agonist dependent and sexually dimorphic[J]. J Pain,2015,16(11):1200-1210.
[18]李云庆.内吗啡肽在脊髓背角通过突触前机制发挥镇痛作用[C]//中华医学会疼痛学分会年会暨casp成立二十周年.2009.
[19] Sakurada S,Hayashi T,Yuhki M,et al. Differential antagonism of endomorphin-1 and endomorphin-2supraspinal antinociception by naloxonazine and 3-methylnaltrexone[J]. Peptides,2002,23(5):895-901.
[20] Delafoy L,Gelot A,Ardid D,et al. Interactive involvement of brain derived neurotrophic factor,nerve growth factor, and calcitonin gene related pep tide in colonic hypersensitivity in the rat[J].Gut,2006,55(7):940-945.
[21] Greenwell TN,Martin-schild S,Inglis FM,et al.Colocalization and shared distribution of endomorphins with substance P,calcitonin gene-related peptide,gamma-aminobutyric acid,and the mu opioid receptor[J]. J Comp Neurol,2007,503(2):319-333.
[22] Sanderson Nydahl K,Skinner K,Julius D,et al. Colocalization of endomorphin-2 and substance P in primary afferent nociceptors and effects of injury:a light and electron microscopic study in the rat[J].Eur J Neurosci,2004,19(7):1789-1799.
[23] Xie H,Woods JH,Traynor JR,et al. The spinal antinociceptive effects of endomorphins in rats:behavioral and G protein functional studies[J]. Anesth Analg,2008,106(6):1873-1881.