神经根型颈椎病动物模型研究进展

神经根型颈椎病(cervical spondylotic radiculopathy, CSR)是由颈椎小关节紊乱或颈椎间盘退行性改变而压迫神经根的一种疾病，典型症状表现为神经根支配区的颈肩部和上肢放射痛或麻木，严重影响着患者的生活质量。为深入研究该疾病病因病机、发展过程、健康防护和药物治疗等，正确地选择和建立符合疾病临床特征的动物模型是解决问题至关重要的环节之一。本文查询国内外相关文献报道，将CSR动物模型的制备归纳总结为单纯压迫模型(血管钳钳夹法、丝线结扎法、L形钢柱压迫法、硅胶片压迫法、自体骨压迫法、尼龙鱼线压迫法)、联合刺激模型(钳夹法联合铬制肠线压迫法、福尔马林定量滤纸刺激法、药物介导联合动静失衡法)和无创性干预模型(低头屈曲固定法)三类。将CSR动物模型制备进行总结概述，分析其特点和优劣势，以期为临床前研究选择适宜的动物模型和探索理想模型提供重要参考依据。     

瞬时外向钾电流的降低介导了慢性压迫背根神经节细胞兴奋性的升高

慢性压迫大鼠背根神经节（chronic compression of the dorsal root,ganglion,CCD）后,背根神经节细胞兴奋性升高,但引起神经元兴奋性改变的离子通道机制还需进一步探索。本实验采用胞内记录以及全细胞膜片钳记录方法,研究急性分离的大鼠背根神经节细胞兴奋性改变与瞬时外向钾电流（A-type potassium current,ⅠA）的关系。结果表明,CCD术后背根神经节细胞兴奋性升高,在急性分离的体外细胞中仍继续存在,表现为对辣椒素敏感的背根神经节细胞产生动作电位的最小电流刺激强度,即阈电流（current threshold）及阈电位（voltage threshold）降低;给予正常对照组神经元（未压迫损伤）瞬时外向钾通道阻断剂4-氨基吡啶,出现了类似CCD术后兴奋性升高的改变。进一步用两步电压钳方法分离ⅠA,研究CCD术后神经元ⅠA的变化,结果表明,CCD组神经元的ⅠA比对照组神经元ⅠA降低,并且与其阈电位的改变一致。以上结果提示,背根神经节压迫受损后,神经节细胞ⅠA降低可能参与介导了神经节细胞兴奋性的升高。     

糖尿病大鼠膀胱和骶髓背根神经节中NGF的表达与尿流动力学改变的研究

目的：探讨糖尿病大鼠膀胱与骶髓背根神经节（DRG）中神经生长因子（NGF）的表达与尿流动力学改变。方法：建立糖尿病大鼠模型10只,对照组10只,应用酶联免疫吸附试验（ELISA）法,分别检测大鼠膀胱组织及骶髓DRG中NGF的变化情况,结合代谢笼及尿流动力学改变,探讨糖尿病膀胱病变的可能发病机制。结果：造模12周后,糖尿病大鼠膀胱容量较正常对照组明显增大（1.47±0.28vs0.71±0.12,p〈0.05）,残余尿量明显增多（0.52±0.18vs0.07±0.08,p〈0.01）,排尿效率明显下降,膀胱及骶髓DRG中NGF表达水平明显降低。结论：NGF在糖尿病大鼠膀胱和骶髓背根神经节中低表达,在糖尿病膀胱病变中发挥着重要作用。     

糖尿病大鼠膀胱和骶髓背根神经节中NGF 的表达与尿流动力学 改变的研究

目的:探讨糖尿病大鼠膀胱与骶髓背根神经节(DRG)中神经生长因子(NGF)的表达与尿流动力学改变。方法:建立糖尿病大鼠模型10只,对照组10只,应用酶联免疫吸附试验(ELISA)法,分别检测大鼠膀胱组织及骶髓DRG中NGF的变化情况,结合代谢笼及尿流动力学改变,探讨糖尿病膀胱病变的可能发病机制。结果:造模12周后,糖尿病大鼠膀胱容量较正常对照组明显增大(1.47±0.28vs0.71±0.12,p<0.05),残余尿量明显增多(0.52±0.18vs0.07±0.08,p<0.01),排尿效率明显下降,膀胱及骶髓DRG中NGF表达水平明显降低。结论:NGF在糖尿病大鼠膀胱和骶髓背根神经节中低表达,在糖尿病膀胱病变中发挥着重要作用。     

慢性炎症痛引起DRG感觉神经元P2X3受体表达的改变

目的探讨脊髓背根神经节（dorsal root ganglia,DRG）P2X3受体参与大鼠足底慢性炎症痛相关的热痛觉过敏机制。方法 1）用行为学的研究方法,以大鼠右侧后脚掌注射松节油加石蜡（各占50％）0．1ml建立后脚掌慢性痛模型,用热测痛的方法测量后脚掌皮下注射松节油后的痛阈,每天1次,连续测15d。2）用免疫组织化学技术观察大鼠后脚掌慢性炎症后第2天和第7天,炎症侧脊髓背根神经节（L4—6）神经元中P2X,受体阳性细胞类型的分布变化;以及正常脊髓背根神经节（L4—6）神经元中P2X,受体阳性细胞类型的分布作为对照。结果1）炎症后大鼠后脚掌侧痛阈出现降低,在第2天痛阈达到最低,后逐渐恢复,14d后恢复正常痛阈值。2）正常大鼠P2X,主要表达于DRG的中小神经元上,炎症后DRG（L4—6）,中小型P2X,受体阳性细胞数比对照组明显增加。细胞平均面积增大。结论后脚掌慢性炎症痛可以引起大鼠对伤害性热刺激的痛觉过敏,并导致脊髓背根神经节（L4—6）神经元qbP2X3受体阳性细胞数目增加,表明P2X3在DRG的中小神经元的改变可能对松节油引起脚掌炎症痛时热痛觉过敏的形成与维持起重要作用。     

大鼠内毒素血症不同时期血浆CGRP和背根神经节CGRP mRNA水平的变化

本文采用逆转录聚合酶链反应（RT－PCR）方法测定大鼠内毒素血症不同时期胸腰段背根神经节降钙素基因相关肽（CGRP）mRNA水平的改变,结合血浆CGRP水平的改变,以期全面了解大鼠内毒素血症不同时期CGRP释放与合成的变化。结果显示：注射内毒素（5mg／kg）后30min时,大鼠血浆CGRP开始增高,而背根神经节CGRPmRNA水平无明显变化;注射内毒素后3h时,血浆CGRP及背根神经节CGRPmRNA均明显增高．分别为142％和32％,8h时则进一步增高,分别为216％和85％。提示内毒素不仅刺激外周组织释放CGRP,而且还能通过某些机制激活背根神经节CGRPmRNA的转录,使CGRP合成增加,以作为CGRP大量释放的重要补充来源。     

电压门控钠通道NaV1.7与痛信号的产生

电压门控钠通道NaV1.7选择性高表达在伤害感受性脊髓背根神经节的感觉神经元上,在疼痛电信号的产生、传导和调控中具有重要的生理功能。伤害性感受器上的NaV1.7亦在慢性神经痛和炎症痛的病理生理过程中发挥关键作用。近年来的研究发现,人类遗传性疼痛症(如红斑性肢痛病)与NaV1.7钠离子通道基因SCN9A的某些功能增强型突变相关。最近Cox等首次报道了SCN9A突变将导致人先天痛觉完全丧失,而无痛症患者机体其它功能正常,提示NaV1.7将可能成为有效治疗疼痛而无副作用的一个新靶标。     

一氧化氮对大鼠背根神经节神经元膜电位的作用

目的：观察一氧化氮对大鼠背根神经节神经元的作用及有关离子机制,并探讨一氧化氮在痛觉信息传递过程中的作用。方法：在分离的大鼠背根神经节标本上,应用细胞内记录技术,给予灌流一氧化氮供体硝普钠,观察硝普钠诱导的神经元膜反应。结果：大部分神经元对硝普钠敏感（79／102,77．45％）,滴加硝普钠（10～100mmol／L）后可引起浓度依赖性的超极化反应,剩余神经元没有反应。硝普钠（100mmol／L）可使神经元膜电导由（21．06±1．94）nS增加到（23．08±0．92）nS。L-NAME（非选择性一氧化氮合酶抑制剂,1mmol／L）、CdCl2（非选择性钙通道阻断剂,0．1mmol／L）、无Na^＋平衡盐液对硝普钠引起超极化反应无明显影响。四乙基碘化铵（非选择性钾通道阻断剂,10mmol／L）明显抑制硝普钠引起的超极化反应。结论：硝普钠在大鼠背根神经节神经元上可引起浓度依赖的超极化反应,且此超极化反应是钾电导介导的。     

子宫内膜异位症大鼠模型的建立及其病理学改变

目的构建子宫内膜异位症（内异症）大鼠动物模型,为阐明内异位症发病机理以及寻找有效的治疗方法提供理想的动物模型。方法取性成熟雌性Sprague-Dawley（SD）大鼠30只,通过手术将大鼠自体子宫组织移植到子宫旁韧带上,建立诱发型内异症大鼠动物模型。术后8周,再次剖腹观察异位组织的存活情况、病灶大小、与周围组织的粘连程度以及病理学变化。结果25只大鼠有明显的异位病灶。所有病灶都与周围组织有不同程度的粘连,病灶外观呈囊泡状。光镜观察见大部分异位子宫内膜形态和结构与在位子宫内膜基本相同,但内膜细胞、间质细胞、腺体,与在位内膜相比较少。少数病灶只有上皮组织或只有问质组织。结论自体子宫移植法可成功建立内异症大鼠模型。     

脑源性神经营养因子在大鼠后足切割疼痛中的作用

目的:观察脑源性神经营养因子(BDNF)对大鼠后足切割疼痛的影响。方法:采用纵行切割大鼠后足作为疼痛模型,运用免疫组织化学与免疫荧光双标记方法,观察大鼠后足切割后不同时间点(1-72hr)BDNF在相应节段背根神经节与脊髓内表达的变化。腹腔或鞘内注射BDNF抗体中和内源性BDNF后,以Von Frey尼龙纤维刺激后足行机械痛敏评价。结果:大鼠后足切割后1-24hr内,BDNF在切割侧L42-L5脊髓后角表达明增加,BDNF主要位于后角神经元内与神经末梢,星形胶质细胞与小胶质细胞内未见明显表达;在L42-L5背根神经节,BNDF免疫阳性细胞百分比在切割后1-24hr内也明显增加,增加的主要为大直经神经元;鞘内给予BDNF抗体可明显增加大鼠后足切割后的缩足阈值,而腹腔给予BDNF抗体对大鼠的缩足阈值影响较小。结论:BDNF参与了大鼠后足切割后机械痛敏的过程。     

Nucleolytic action of chondroitinase ABC on the lumbar disc of the rabbit

Nucleolysis using chondroitinase ABC was studied using the rabbit's intervertebral lumbar disc. The purpose was to find a possible alternative to chymopapain which is commonly used in the management of sciatica due to disc herniation. The injection of 1 U of the enzyme into the nucleus pulposus gave significant histological and biochemical changes in all twelve discs studied.     

椎间孔镜下髓核摘除术对腰椎间盘突出患者CK、CRP及腰椎功能恢复的影响

目的:探讨椎间孔镜下髓核摘除术对椎间盘突出患者肌酸激酶(CK)、C反应蛋白(CRP)及腰椎功能恢复的影响。方法:选择2016年10月-2018年8月在我院接受治疗的90例腰椎间盘突出患者,采用抽签法分为椎间孔镜组(n=45)和后路切开组(n=45)。对照组给予后路切开髓核摘除手术治疗,观察组给予椎间孔镜下髓核摘除治疗。比较两组患者的手术情况、CK、CRP、腰椎功能、视觉模拟(VAS)评分、Oswestry功能障碍指数(ODI)评分变化情况及并发症发生情况。结果:椎间孔镜组手术时间较后路切开组更长,术中出血量、术后下床时间及住院时间较后路切开组更低(P<0.05);手术前,两组CK、CRP检测结果无差异;手术后,两组CK、CRP均随着时间的延长均呈上升趋势,且椎间孔镜组上升较后路切开组低(P<0.05);手术前,两组腰椎功能检测结果无差异;手术后,两组腰椎曲度、直腿抬高试验均随着时间的延长均呈上升趋势,且椎间孔镜组上升更为明显(P<0.05);手术前,两组VAS、ODI评分评定结果无差异;手术后,两组VAS、ODI评分均随着时间的推移均呈下降趋势,且椎间孔镜组下降更为明显(P<0.05);手术后,与后路切开组24.44%(11/45)进行比较,椎间孔镜组4.44%(2/45)显著降低(P<0.05)。结论:在腰椎间盘突出症患者中应用椎间孔镜下髓核摘除效果显著,可有效改善CK、CRP及腰椎功能水平。     

Mechanical thresholds for initiation and persistence of pain following nerve root injury: mechanical and chemical contributions at injury

There is much evidence supporting the hypothesis that magnitude of nerve root mechanical injury affects the nature of the physiological responses which can contribute to pain in lumbar radiculopathy. Specifically, injury magnitude has been shown to modulate behavioral hypersensitivity responses in animal models of radiculopathy. However, no study has determined the mechanical deformation thresholds for initiation and maintenance of the behavioral sensitivity in these models. Therefore, it was the purpose of this study to quantify the effects of mechanical and chemical contributions at injury on behavioral outcomes and to determine mechanical thresholds for pain onset and persistence. Male Holtzman rats received either a silk or chromic gut ligation of the L5 nerve roots, a sham exposure of the nerve roots, or a chromic exposure in which no mechanical deformation was applied but chromic gut material was placed on the roots. Using image analysis, nerve root radial strains were estimated at the time of injury. Behavioral hypersensitivity was assessed by measuring mechanical allodynia continuously throughout the study. Chromic gut ligations produced allodynia responses for nerve root strains at two-thirds of the magnitudes of those strains which produced the corresponding behaviors for silk ligation. Thresholds for nerve root compression producing the onset (8.4%) and persistence of pain (17.4%-22.2%) were determined for silk ligation in this lumbar radiculopathy model. Such mechanical thresholds for behavioral sensitivity in a painful radiculopathy model begin to provide biomechanical data which may have utility in broader experimental and computational models for relating injury biomechanics and physiologic responses of pain.     

PeroxiredoxinⅡ在退变椎间盘髓核中的表达及其临床意义

目的：检测PeroxiredoxinⅡ在腰椎间盘髓核组织中的表达,分析其在椎间盘退变中的临床意义。方法：用蛋白免疫印迹（Western blot）的方法检测PeroxiredoxinⅡ在正常、突出及脱出腰椎间盘髓核中的表达情况。结果：PeroxiredoxinⅡ在退变椎间盘髓核中表达丰富,而在正常椎间盘髓核中表达微弱,两者比较差异显著（P〈0.05）,在突出和脱出椎间盘髓核中表达无显著性差异（P〉0.05）。结论：PeroxiredoxinⅡ在正常及退变腰椎间盘髓核组织中差异表达。     

Numerical analysis of the influence of nucleus pulposus removal on the biomechanical behavior of a lumbar motion segment

Nucleus replacement was deemed to have therapeutic potential for patients with intervertebral disc herniation. However, whether a patient would benefit from nucleus replacement is technically unclear. This study aimed to investigate the influence of nucleus pulposus (NP) removal on the biomechanical behavior of a lumbar motion segment and to further explore a computational method of biomechanical characteristics of NP removal, which can evaluate the mechanical stability of pulposus replacement. We, respectively, reconstructed three types of models for a mildly herniated disc and three types of models for a severely herniated disc based on a L4–L5 segment finite element model with computed tomography image data from a healthy adult. First, the NP was removed from the herniated disc models, and the biomechanical behavior of NP removal was simulated. Second, the NP cavities were filled with an experimental material (Poisson's ratio = 0.3; elastic modulus = 3 MPa), and the biomechanical behavior of pulposus replacement was simulated. The simulations were carried out under the five loadings of axial compression, flexion, lateral bending, extension, and axial rotation. The changes of the four biomechanical characteristics, i.e. the rotation degree, the maximum stress in the annulus fibrosus (AF), joint facet contact forces, and the maximum disc deformation, were computed for all models. Experimental results showed that the rotation range, the maximum AF stress, and joint facet contact forces increased, and the maximum disc deformation decreased after NP removal, while they changed in the opposite way after the nucleus cavities were filled with the experimental material.     

3.0T磁共振扩张量成像对腰椎间盘突出致神经根受压的诊断价值及其与Oswestry功能障碍指数及视觉模拟评分的相关性

目的:研究3.0T磁共振扩张量成像(DTI)对腰椎间盘突出致神经根受压的诊断价值及其与Oswestry功能障碍指数(ODI)及视觉模拟评分(VAS)的相关性。方法:纳入我院从2017年1月~2019年1月收治的腰椎间盘突出致神经根受压患者50例进行研究,记作研究组。另取同期我院收治的单纯腰椎间盘突出患者50例作为对照组。两组受试者均接受DTI扫描以及ODI、VAS评分。比较两组神经根不同层面的各向异性分数(FA)值、弥散系数(ADC)值、ODI、VAS评分,并作相关性分析。同时,以手术病理诊断为金标准,分析DTI诊断腰椎间盘突出致神经根受压的敏感性、特异性、准确度。结果:研究组患者神经根近层、中层、远层的FA值均显著低于对照组,而ADC值均显著高于对照组(均P<0.05)。以手术病理诊断为金标准,DTI诊断腰椎间盘突出致神经根受压的敏感性为94.00%、特异性为96.00%、准确度为95.00%。研究组ODI、VAS评分分别为(43.22±7.25)分、(6.68±1.92)分,相较于对照组的(28.56±6.22)分、(4.02±1.34)分显著更高(均P<0.05)。经Pearson相关性分析可得:腰椎间盘突出致神经根受压患者的FA值与ODI、VAS评分均呈负相关关系(均P<0.05),而ADC值与ODI、VAS评分无相关性(均P>0.05)。结论:DTI对腰椎间盘突出致神经根受压的诊断价值较高,且FA值与ODI、VAS均存在明显相关性。临床工作中可能将DTI的FA值作为量化神经根结构改变的重要参数,值得临床重点关注。     

正常椎间盘胶原的研究

腰椎间盘突出症是引起腰腿痛常见的原因。胶原作为椎间盘结构的主要成分,构成椎间盘的纤维框架,其类型与分布直接决定着椎间盘结构的强度和功能的稳定。本文利用溴化氰消化椎间盘胶原产生多肽,借助于梯度层析。SDS-PAGE及光密度定量扫描等对正常人椎间盘胶原进行了研究。结果表明:正常人椎间盘含Ⅰ型及Ⅱ型两种胶原,它们的分布呈明显而特征性的移行性变化:纤维环外层边缘以Ⅰ型胶原为主(83％),由外向内Ⅰ型胶原逐渐移行为Ⅱ型胶原,靠近髓核处以Ⅱ型胶原为主(72％);髓核中心含有Ⅱ型胶原。此为椎间盘的一个结构特性,以满足椎间盘的特殊功能的需要。     

Changes in the intervertebral disks in disorders of segmental blood supply of the spine

In 50 mature Chinchilla rabbits a model of chronic insufficiency of blood supply in the lumbar vertebral bodies has been disturbed as a result of unilateral sectioning of the segmentary arteries and veins. By means of light and transmissive electron microscopy the dynamics of structural changes has been followed in tissue of the intervertebral discs for 3 months after the operative intervention. Under hypoxia in the ground substance of the pulposus++ nucleus even proteoglycans granular-filamentous network gradually develops and floccular material and transverse striated filamentous aggregates are accumulated. Notochord cells are subjected to certain degenerative changes and die. Simultaneously fibroblastic cells of the pulposus++ nucleus periphery become activated, they produce glycosaminoglycans and collagen. As a result the hydrated tissue of the pulposus++ nucleus is substituted for a newly formed fibrous cartilage. The process of fibroses in the intervertebral disc is completed in 3 months after blood circulation has been disturbed in the vertebral bodies.     

中医针灸推拿配合牵引治疗对老年人腰椎间盘突出症的临床效果

目的：探讨中医针灸推拿配合牵引治疗在老年人腰椎间盘突出症的临床效果。方法：将我院2010年5月至2012年12月间收治的182例老年腰椎间盘突出症患者随机分为综合治疗组（91例）和对照组（91例）,对照组接受牵引治疗,综合治疗组采用针灸推拿配合牵引治疗。观察比较两组患者治疗前后的症状改善及临床疗效。结果：经过3周治疗,综合治疗组的患者总有效率为95．6％,对照组的患者总有效率为60．40／0,其差异具有统计学意义（P〈0．05）。综合治疗组治疗后FIM评分较治疗前相比各项均有明显改善,其结果具有统计学差异（P〈O．05）,对照组除括约肌控制及社会交流和认可方面无改善。其余项目FIM评分较治疗前有显著提高（P〈0．05）。综合治疗组治疗后FIM评分的提高显著高于对照组治疗后的评分（P〈0．05）。结论：中医针灸推拿配合牵引治疗对老年人腰椎间盘突出症的疗效显著,值得在临床推广应用。     

Measurement of in vivo lumbar intervertebral disc pressure during spinal manipulation: a feasibility study

This paper presents the first reported measurements of lumbar intervertebral disc pressure in vivo during spinal manipulation. A pressure transducer was inserted into the nucleus pulposus of one normal-appearing lumbar disc in an asymptomatic adult volunteer. Pressures were recorded during several body positions and maneuvers, then during spinal manipulation, and lastly during a repetition of the preintervention body positions. Baseline pressures in the prone and side-lying positions measured 110 kPa and 150 kPa, respectively. During the manipulation, pressure rose to a peak of 890 kPa over 250 ms. Immediately following, pressures in the prone and side-lying positions measured 150 kPa and 165 kPa, respectively. These data do not support the hypotheses that manipulation can reduce a herniation by decreasing intradiscal pressure, or cause a herniation by raising pressure to failure levels. Further work may lead to a better understanding of this treatment method.