神经肽Y和受体在腰椎动静力失衡大鼠模型中表达及对痛阈的影响
|
摘要
目的:观察神经肽Y(neuropeptide Y,NPY)及Y1R、Y2R在椎间盘退变(intervertebral disc degeneration,IDD)组和假手术(sham operation,SHAM)组大鼠椎间盘(intervertebral disc,IVD)和背根神经节(dorsal root ganglion,DRG)表达的差异,以及与大鼠机械缩足阈值(mechanical withdrawal threshold,MWT)和热缩足反射潜伏期(thermal withdrawal latency,TWL)的相关关系,探讨NPY及受体与椎间盘源性腰痛的关系。方法:选择6月龄无特定病原体(specific pathogen free,SPF)级雌性未孕SD大鼠12只,随机分为椎间盘退变组和假手术组(n=6),前者采用动静力失衡大鼠椎间盘退变模型,后者做假手术处理,造模后12周通过MRI和X线评估椎间盘退变模型建立是否成功并测量2组大鼠MWT及TWL,使用免疫组织化学法检测2组大鼠IVD和DRG中NPY及受体的表达,分析其平均光密度(mean optical density,MOD)值与疼痛行为学之间的相关性。结果:(1)疼痛检测发现,术后12周IDD组大鼠MWT较SHAM组大鼠明显降低[(9.44±2.91)g vs.(17.99±4.40)g,P=0.002],TWL明显降低[(9.17±0.39)s vs.(13.53±1.69)s,P=0.002]。(2)免疫组织化学法检测显示,与SHAM组相比,IDD组大鼠IVD和DRG中NPY的表达明显升高[IVD:[(0.15±0.03)vs.(0.11±0.02),P=0.021;DRG:(0.23±0.04)vs.(0.17±0.04),P=0.018],Y1R表达明显降低[IVD:(0.10±0.02)vs.(0.13±0.01),P=0.009;DRG:(0.05±0.01)vs.(0.08±0.01),P=0.000],2组大鼠IVD和DRG中Y2R的表达差异均无统计学意义。(3)Pearson相关性分析表明,2组大鼠IVD和DRG中NPY的MOD值与MWT之间成负相关关系(r=-0.605,P=0.037;r=-0.617,P=0.033);2组大鼠IVD和DRG中Y1R的MOD值与MWT之间成正相关关系(r=0.696,P=0.012;r=0.735,P=0.006)。2组大鼠IVD和DRG中NPY的MOD值与TWL之间成负相关关系(r=-0.603,P=0.038;r=-0.708,P=0.010);2组大鼠IVD和DRG中Y1R的MOD值与TWL之间成正相关关系(r=0.624,P=0.030;r=0.834,P=0.001)。结论:在腰椎动静力失衡大鼠模型中,NPY可能通过外周Y1R参与椎间盘源性腰痛的发生发展,但其具体机制有待进一步研究。
Objective:To investigate the expression of neuropeptide Y(NPY),Y1 receptor(Y1 R),and Y2 receptor(Y2 R)in the intervertebral disc(IVD)and dorsal root ganglion(DRG)of rats with intervertebral disc degeneration(IDD)and sham-operated rats,and to investigate the correlation of NPY and its receptors with mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)in rats and the relationship of NPY and its receptors with discogenic low back pain. Methods:Twelve 6-month-old female specific pathogen-free Sprague-Dawley rats which were not pregnant were equally and randomly divided into two groups:IDD group and sham-operation group. An IDD model was induced by unbalanced dynamic and static forces in the IDD group,and sham operation was performed in the sham-operation group. At 12 weeks after the operation,whether the establishment of the IDD model was successful was evaluated by magnetic resonance imaging and X-ray,and MWT and TWL were measured. The expression of NPY and its receptors in the IVD and DRG was determined by immunohistochemistry to analyze the correlation of the mean optical density(MOD)of NPY and its receptors with pain behaviors(MWT and TWL). Results:(1)Pain testing results showed that the IDD group had significantly lower MWT and TWL compared with the sham-operation group at 12 weeks after operation[(9.44 ±2.91) g vs.(17.99±4.40) g,P=0.002;(9.17±0.39) s vs.(13.53 ±1.69) s,P =0.002].(2) Immunohistochemical analysis showed that the IDD group had significantly higher expression of NPY in the IVD and DRG and significantly lower expression of Y1 R in the IVD and DRG compared with the sham-operation group(IVD:0.15±0.03 vs. 0.11±0.02,P=0.021;DRG:0.23±0.04 vs.0.17±0.04,P=0.018;IVD:0.10±0.02 vs. 0.13±0.01,P=0.009;DRG:0.05±0.01 vs. 0.08±0.01,P=0.000). There was no significant difference in the expression of Y2 R in the IVD and DRG between the two groups(P>0.05).(3)Pearson correlation analysis showed that the MOD values of NPY in the IVD and DRG were negatively correlated with MWT and TWL in the two groups(r=-0.605,P=0.037;r=-0.617,P=0.033;r=-0.603,P=0.038;r=-0.708,P=0.010);the MOD values of Y1 R in the IVD and DRG were positively correlated with MWT and TWL in the two groups(r=0.696,P=0.012;r=0.735,P=0.006;r=0.624,P=0.030;r=0.834,P=0.001).Conclusion:In the rat model of unbalanced dynamic and static forces,NPY may be involved in the development and progression of discogenic low back pain through peripheral Y1 R,but the specific mechanism needs further investigations.
Objective:To investigate the expression of neuropeptide Y(NPY),Y1 receptor(Y1 R),and Y2 receptor(Y2 R)in the intervertebral disc(IVD)and dorsal root ganglion(DRG)of rats with intervertebral disc degeneration(IDD)and sham-operated rats,and to investigate the correlation of NPY and its receptors with mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)in rats and the relationship of NPY and its receptors with discogenic low back pain. Methods:Twelve 6-month-old female specific pathogen-free Sprague-Dawley rats which were not pregnant were equally and randomly divided into two groups:IDD group and sham-operation group. An IDD model was induced by unbalanced dynamic and static forces in the IDD group,and sham operation was performed in the sham-operation group. At 12 weeks after the operation,whether the establishment of the IDD model was successful was evaluated by magnetic resonance imaging and X-ray,and MWT and TWL were measured. The expression of NPY and its receptors in the IVD and DRG was determined by immunohistochemistry to analyze the correlation of the mean optical density(MOD)of NPY and its receptors with pain behaviors(MWT and TWL). Results:(1)Pain testing results showed that the IDD group had significantly lower MWT and TWL compared with the sham-operation group at 12 weeks after operation[(9.44 ±2.91) g vs.(17.99±4.40) g,P=0.002;(9.17±0.39) s vs.(13.53 ±1.69) s,P =0.002].(2) Immunohistochemical analysis showed that the IDD group had significantly higher expression of NPY in the IVD and DRG and significantly lower expression of Y1 R in the IVD and DRG compared with the sham-operation group(IVD:0.15±0.03 vs. 0.11±0.02,P=0.021;DRG:0.23±0.04 vs.0.17±0.04,P=0.018;IVD:0.10±0.02 vs. 0.13±0.01,P=0.009;DRG:0.05±0.01 vs. 0.08±0.01,P=0.000). There was no significant difference in the expression of Y2 R in the IVD and DRG between the two groups(P>0.05).(3)Pearson correlation analysis showed that the MOD values of NPY in the IVD and DRG were negatively correlated with MWT and TWL in the two groups(r=-0.605,P=0.037;r=-0.617,P=0.033;r=-0.603,P=0.038;r=-0.708,P=0.010);the MOD values of Y1 R in the IVD and DRG were positively correlated with MWT and TWL in the two groups(r=0.696,P=0.012;r=0.735,P=0.006;r=0.624,P=0.030;r=0.834,P=0.001).Conclusion:In the rat model of unbalanced dynamic and static forces,NPY may be involved in the development and progression of discogenic low back pain through peripheral Y1 R,but the specific mechanism needs further investigations.
引文
[1] Andersson GB. Epidemiological features of chronic low-back pain[J]. Lancet,1999,354(9178):581-585.
[2] Petersen T,Laslett M,Juhl C. Clinical classification in low back pain:best-evidence diagnostic rules based on systematic reviews[J]. BMC Musculoskelet Disord,2017,18(1):188.
[3] McCarron RF,Wimpee MW,Hudkins PG,et al. The inflammatory effect of nucleus pulposus. A possible element in the pathogenesis of low-back pain[J]. Spine(Phila Pa 1976),1987,12(8):760-764.
[4] Schroeder M,Viezens L,Schaefer C,et al. Chemokine profile of disc degeneration with acute or chronic pain[J]. J Neurosurg Spine,2013,18(5):496-503.
[5] Krock E,Rosenzweig DH,Chabot-Dore AJ,et al. Painful,degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors[J]. J Cell Mol Med,2014,18(6):1213-1225.
[6] Barczewska M,Juranek J,Wojtkiewicz J. Origins and neurochemical characteristics of porcine intervertebral disc sympathetic innervation:a preliminary report[J]. J Mol Neurosci,2017,63(1):50-57.
[7] Zhang K,Ding W,Sun W,et al. Beta1 integrin inhibits apoptosis induced by cyclic stretch in annulus fibrosus cells via ERK1/2 MAPK pathway[J]. Apoptosis,2016,21(1):13-24.
[8] Chaplan SR,Bach FW,Pogrel JW,et al. Quantitative assessment of tactile allodynia in the rat paw[J]. J Neurosci Methods,1994,53(1):55-63.
[9] Geurts JW,Willems PC,Kallewaard JW,et al. The impact of chronic discogenic low back pain:costs and patients’burden[J]. Pain Res Manag,2018,2018:4696180.
[10] Miyagi M,Millecamps M,Danco AT,et al. ISSLS Prize winner:increased innervation and sensory nervous system plasticity in a mouse model of low back pain due to intervertebral disc degeneration[J]. Spine(Phila Pa 1976),2014,39(17):1345-1354.
[11] Intondi AB,Zadina JE,Zhang X,et al. Topography and time course of changes in spinal neuropeptide Y immunoreactivity after spared nerve injury[J]. Neuroscience,2010,165(3):914-922.
[12] Lai A,Moon A,Purmessur D,et al. Annular puncture with tumor necrosis factor-alpha injection enhances painful behavior with disc degeneration in vivo[J]. Spine J,2016,16(3):420-431.
[13] Shi TJ,Li J,Dahlstrom A,et al. Deletion of the neuropeptide Y Y1 receptor affects pain sensitivity,neuropeptide transport and expression,and dorsal root ganglion neuron numbers[J]. Neuroscience,2006,140(1):293-304.
[14] Taylor BK,Fu W,Kuphal KE,et al. Inflammation enhances Y1receptor signaling,neuropeptide Y-mediated inhibition of hyperalgesia,and substance P release from primary afferent neurons[J]. Neuroscience,2014,256:178-194.
[15] Robich MP,Matyal R,Chu LM,et al. Effects of neuropeptide Y on collateral development in a swine model of chronic myocardial ischemia[J]. J Mol Cell Cardiol,2010,49(6):1022-1030.
[16] Magnussen C,Hung SP,Ribeiro-da-Silva A. Novel expression pattern of neuropeptide Y immunoreactivity in the peripheral nervous system in a rat model of neuropathic pain[J]. Mol Pain,2015,11:31.
[17] Shi TJ,Zhang X,Berge OG,et al. Effect of peripheral axotomy on dorsal root ganglion neuron phenotype and autonomy behaviour in neuropeptide Y-deficient mice[J]. Regul Pept,1998,75-76:161-173.
[18] Navone SE,Marfia G,Giannoni A,et al. Inflammatory mediators and signalling pathways controlling intervertebral disc degeneration[J].Histol Histopathol,2017,32(6):523-542.
[19] Ji RR,Zhang X,Wiesenfeld-Hallin Z,et al. Expression of neuropeptide Y and neuropeptide Y(Y1)receptor mRNA in rat spinal cord and dorsal root ganglia following peripheral tissue inflammation[J]. J Neurosci,1994,14(11 Pt 1):6423-6434.
[20] Iwagaki N,Ganley RP,Dickie AC,et al. A combined electrophysiological and morphological study of neuropeptide Y-expressing inhibitory interneurons in the spinal dorsal horn of the mouse[J]. Pain,2016,157(3):598-612.
[2] Petersen T,Laslett M,Juhl C. Clinical classification in low back pain:best-evidence diagnostic rules based on systematic reviews[J]. BMC Musculoskelet Disord,2017,18(1):188.
[3] McCarron RF,Wimpee MW,Hudkins PG,et al. The inflammatory effect of nucleus pulposus. A possible element in the pathogenesis of low-back pain[J]. Spine(Phila Pa 1976),1987,12(8):760-764.
[4] Schroeder M,Viezens L,Schaefer C,et al. Chemokine profile of disc degeneration with acute or chronic pain[J]. J Neurosurg Spine,2013,18(5):496-503.
[5] Krock E,Rosenzweig DH,Chabot-Dore AJ,et al. Painful,degenerating intervertebral discs up-regulate neurite sprouting and CGRP through nociceptive factors[J]. J Cell Mol Med,2014,18(6):1213-1225.
[6] Barczewska M,Juranek J,Wojtkiewicz J. Origins and neurochemical characteristics of porcine intervertebral disc sympathetic innervation:a preliminary report[J]. J Mol Neurosci,2017,63(1):50-57.
[7] Zhang K,Ding W,Sun W,et al. Beta1 integrin inhibits apoptosis induced by cyclic stretch in annulus fibrosus cells via ERK1/2 MAPK pathway[J]. Apoptosis,2016,21(1):13-24.
[8] Chaplan SR,Bach FW,Pogrel JW,et al. Quantitative assessment of tactile allodynia in the rat paw[J]. J Neurosci Methods,1994,53(1):55-63.
[9] Geurts JW,Willems PC,Kallewaard JW,et al. The impact of chronic discogenic low back pain:costs and patients’burden[J]. Pain Res Manag,2018,2018:4696180.
[10] Miyagi M,Millecamps M,Danco AT,et al. ISSLS Prize winner:increased innervation and sensory nervous system plasticity in a mouse model of low back pain due to intervertebral disc degeneration[J]. Spine(Phila Pa 1976),2014,39(17):1345-1354.
[11] Intondi AB,Zadina JE,Zhang X,et al. Topography and time course of changes in spinal neuropeptide Y immunoreactivity after spared nerve injury[J]. Neuroscience,2010,165(3):914-922.
[12] Lai A,Moon A,Purmessur D,et al. Annular puncture with tumor necrosis factor-alpha injection enhances painful behavior with disc degeneration in vivo[J]. Spine J,2016,16(3):420-431.
[13] Shi TJ,Li J,Dahlstrom A,et al. Deletion of the neuropeptide Y Y1 receptor affects pain sensitivity,neuropeptide transport and expression,and dorsal root ganglion neuron numbers[J]. Neuroscience,2006,140(1):293-304.
[14] Taylor BK,Fu W,Kuphal KE,et al. Inflammation enhances Y1receptor signaling,neuropeptide Y-mediated inhibition of hyperalgesia,and substance P release from primary afferent neurons[J]. Neuroscience,2014,256:178-194.
[15] Robich MP,Matyal R,Chu LM,et al. Effects of neuropeptide Y on collateral development in a swine model of chronic myocardial ischemia[J]. J Mol Cell Cardiol,2010,49(6):1022-1030.
[16] Magnussen C,Hung SP,Ribeiro-da-Silva A. Novel expression pattern of neuropeptide Y immunoreactivity in the peripheral nervous system in a rat model of neuropathic pain[J]. Mol Pain,2015,11:31.
[17] Shi TJ,Zhang X,Berge OG,et al. Effect of peripheral axotomy on dorsal root ganglion neuron phenotype and autonomy behaviour in neuropeptide Y-deficient mice[J]. Regul Pept,1998,75-76:161-173.
[18] Navone SE,Marfia G,Giannoni A,et al. Inflammatory mediators and signalling pathways controlling intervertebral disc degeneration[J].Histol Histopathol,2017,32(6):523-542.
[19] Ji RR,Zhang X,Wiesenfeld-Hallin Z,et al. Expression of neuropeptide Y and neuropeptide Y(Y1)receptor mRNA in rat spinal cord and dorsal root ganglia following peripheral tissue inflammation[J]. J Neurosci,1994,14(11 Pt 1):6423-6434.
[20] Iwagaki N,Ganley RP,Dickie AC,et al. A combined electrophysiological and morphological study of neuropeptide Y-expressing inhibitory interneurons in the spinal dorsal horn of the mouse[J]. Pain,2016,157(3):598-612.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |