uPA、MMP-3在退变椎间盘中的表达及意义

目的探讨尿激酶型纤溶酶原激活剂（uPA）、基质金属蛋白-3（MMP-3）在人类退变椎间盘组织中的表达变化。方法对照组为取自脊柱侧凸患者的正常腰椎间盘组织；实验组为腰椎间盘突出患者的退变椎间盘组织。分别进行免疫组织化学染色，比较2组椎间盘组织中uPA及MMP-3的光密度值。结果uPA和MMP-3在突出椎间盘组织中平均光密度值较正常椎间盘组织中的平均光密度值表达明显升高；相关分析显示突出椎间盘中uPA、MMP-3呈正相关关系。结论uPA和MMP-3均可能参与了人椎间盘组织的退变过程，且MMP-3与uPA在人类退变椎间盘组织中有相同的增高趋势。     

退变腰椎间盘组织中碱性成纤维细胞生长因子的表达研究

目的 探讨碱性成纤维细胞生长因子（basic fibroblast growth factor,bFGF)在正常和退变椎间盘组织中的表达情况。方法 交30例来源于腰椎间盘突出症患者手术中所取得的椎间盘组织（观察组，男11例，女19例；年龄25－78岁，平均48岁；病程3个月－30年，平均9年11个月）与6例来源于脊柱侧凸患者前路松解术所取得的椎间盘组织（对照组，男女各3例，年龄10－17岁，平均14.2岁）进行对比，首先经病理组织学检查证实为退变椎间盘组织和政党椎间盘组织，然后将两组椎间盘组织分别通过免疫组织化学方法和原位杂交方法，检测各自椎间盘组织中的bFGF及其mRNA的表达。观察组30例均为退变椎间盘组织，免疫组化阳性率为90％（27／30），原位杂交阳性率为20％（6／30）；对照组6例均为正常椎间盘组织，其免疫组及原位杂交均为阴性，两组间免疫组化方法检测阳性率在统计学上差异有非常显著性意义。结论 bFGF在正常和退变椎间盘组织中表达的差异有显著性意义。提示 bFGF可能作为增生刺激因子促进椎间盘组织中的软骨细胞增生和细胞外基质合成，进而加速椎间盘退变。     

MMP-3、IgG和CD68在青少年与中老年突出腰椎间盘组织中的表达分析

【摘要】 目的：比较细胞因子基质金属蛋白酶3（matrix metalloproteinases 3,MMP-3）、IgG和CD68在青少年与中老年患者突出腰椎间盘组织中的表达,探讨两个年龄段腰椎间盘突出的病因。方法：收集40例腰椎间盘突出症患者的突出腰椎间盘髓核标本,青少年组18例,年龄11～25岁,平均20.6±3.4岁;中老年组22例,年龄40～72岁,平均48.0±10.3岁。HE染色观察标本退变情况,免疫组化染色检测MMP-3、IgG和CD68的表达,光学显微镜下观测并计录数据。结果：标本HE染色提示多数（17/22）中老年组腰椎间盘存在明显退变,青少年组多数椎间盘（13/18）退变不明显或无退变。MMP-3阳性率青少年组（33.33%）低于中老年组（81.82%）;IgG阳性率青少年组（66.67%）高于中老年组（31.82%）;CD-68阳性率青少年组（83.33%）高于中老年组（45.45%）,两组MMP-3、IgG和CD68的表达阳性率比较,差异均有统计学意义（P<0.05）。结论：免疫和炎症反应可能是青少年腰椎间盘突出的重要病因,而中老年腰椎间盘突出可能主要与退变有关。     

新型山羊腰椎间盘突出模型的建立

目的建立新型的山羊腰椎间盘退变合并突出的动物模型。方法9只6月龄山羊，随机分成3组，每组18个腰椎间盘，再将每只山羊的6个腰椎间盘随机等分成正常组和模型组。模型组椎间盘左前外侧纤维环用直径2．5mm的微型环锯限深5mm造成柱状纤维环缺损。术后2、4、6个月时对正常及模型椎间盘摄X线片和MR扫描，观察X线片上的椎间盘高度指数、MRI T2加权像上的椎间盘高信号区的面积和平均R弛豫时间、椎间盘纤维环损伤区的形态学变化，进行组织病理学切片观察（HE染色、Masson三色胶原染色和番红O-固绿染色），对椎间盘组织形态学变化、生物化学变化和组织学评分等指标进行评估。结果正常成年山羊椎间盘组织学结构特点与人类相似；模型椎间盘在损伤后2个月即出现明显退变（P〈0．01），包括椎间盘高度指数降低、椎间盘T2加权像高信号区面积和平均R弛豫时间减低、组织学退变评分增高；MRI和组织学均发现损伤纤维环外侧1／3修复，内2／3出现髓核组织突出充填。损伤后4、6个月模型椎间盘退变加重，但与损伤后2个月的指标差异无统计学意义（P〉0．05）；椎间盘突出形态稳定。结论以成年山羊为对象，应用微型环锯对椎间盘前外侧纤维环定量损伤可以获得快速、有效、重复性好的腰椎间盘突出动物模型。     

TNF-α、IL-1β在退变椎间盘组织中的表达及其意义

目的探讨肿瘤坏死因子-α（TNF—α）、白细胞介素-1β（IL-1β）在人类突出椎间盘组织中的表达变化及意义。方法实验组为手术切除的腰椎间盘突出患者的退变椎间盘组织,分为纤维环破裂组和未破裂组;对照组为取自腰椎骨折患者的正常腰椎间盘组织。采用双抗体夹心ABC—ELISA法分别测定3组椎间盘组织中的TNF-α、IL-1β的含量并进行比较。结果TNF-α和IL-1β含量在纤维环破裂组和纤维环未破裂组均明显高于对照组,纤维环破裂组明显高于纤维环未破裂组（P〈0．05）。结论TNF—α和IL-1β均可能参与了人类椎间盘组织的退变过程,且两者在突出的椎间盘组织中有相同的增高趋势。     

退变腰椎间盘组织的血管浸润现象及其意义

目的：观察退变腰椎间盘的血管浸润现象。方法：对５７例经手术治疗的腰椎间盘突出症患者的退变椎间盘组织及３９个（１３例）正常椎间盘组织做了组织学观察。结果：破裂型突出的血管浸润率（７２．０％）高于凸起型（１８．８％）；病程在一年以内的血管浸润率（５１．２％）高于病程超过一年者（１８．８％），患者年龄与血管浸润无直接关系。结论：腰椎间盘突出的不同病理类型不仅是形态学上的差异，也具有各自不同的组织学基础，破裂型突出的较高血管浸润率很可能与其较严重的临床表现以及较明显的组织退变相关联。血管浸润主要出现在腰椎间盘突出的早期或急性阶段，这不仅是该病变的一种伴随现象，很可能在椎间盘退变中起关键性的始动作用     

环氧化酶2与血管内皮生长因子在突出腰椎间盘中的表达及意义

目的：探讨环氧化酶2（COX-2）、血管内皮生长因子（VEGF）在突出腰椎间盘中的表达及其意义。方法：62个突出椎间盘标本取自58例腰椎间盘突出症手术患者,包括突起型22个,破裂型20个,游离型20个。取材部位分别为突出组织或游离组织（A部位）和椎间隙残余髓核组织（B部位）。对照组取自4例新鲜年轻尸体的L3/4、L4/5及L5/S1椎间盘组织共12个标本,取材部位为椎间盘边缘（A部位）和中央髓核（B部位）;应用免疫组化法对各组标本中COX-2和VEGF的表达进行检测;应用图像分析系统测量标本中COX-2和VEGF表达量的平均光密度值。结果：在腰椎间盘突出组,特别是破裂型和游离型组的突出组织中存在富含新生血管的肉芽组织,COX-2和VEGF染色阳性细胞主要表达于肉芽组织中及突出组织的椎间盘细胞中,对照组未见阳性染色细胞。在突出组的A部位从突起型、破裂型到游离型COX-2和VEGF的表达均逐渐增高,差异有显著性（P〈0.01）。突出组的A部位COX-2和VEGF的表达明显高于B部位（P〈0.01）。腰椎间盘组织中COX-2和VEGF的表达存在明显相关性（r=0.855,P〈0.01）。结论：COX-2、VEGF参与腰椎间盘退变、突出的发病过程;随着腰椎间盘退变突出的进展,COX-2、VEGF的表达均逐渐增高;COX-2与VEGF表达密切相关。     

青少年与中老年腰椎间盘突出相关因素分析

[目的]检测细胞因子基质金属蛋白酶(matrix metalloproteinases,MMPs)MMP-3,IgG和CD68在青少年与中老年患者突出腰椎间盘组织中的表达,探讨两个年龄段腰椎间盘突出的病因.[方法]收集42例腰椎间盘突出症患者的突出腰椎间盘髓核标本,青少年组18例;年龄11 ～25岁,平均(20.59 ±3.39)岁;中老年组24例,年龄30～72岁,平均(47±11.15)岁.HE染色观察标本退变情况,免疫组化检测MMP-3,IgG和CD68的表达,光学显微镜下观测并记录数据.[结果] MMP-3阳性率青少年组(33.3％)低于中老年组(79.2％);IgG阳性率青少年组(66.7％)高于中老年组(29.2％);CD-68阳性率青少年组(83.3％)高于中老年组(54.2％),两组MMP-3,IgG和CD68的表达阳性率比较,均P＜0.05,差异均有统计学意义.标本HE染色提示中老年组腰椎间盘存在明显退变,青少年组椎间盘退变不明显或无退变.[结论]病理表明中老年组突出腰椎间盘组织存在明显退变,而青少年组退变不明显.免疫组化检测显示中老年组MMP-3明显高于青少年组,但青少年组IgG和CD68均明显高于中老年组,免疫和炎症反应可能是青少年腰椎间盘突出的重要病因,而中老年腰椎间盘突出可能主要与退变有关.     

动物模型椎间盘退变全程基因变化的对比

目的：新西兰大白兔的纤维环损伤制作腰椎间盘退变模型，用以证实和比较在人椎间盘退变中明显变化的基因变化情况。方法：2．0只新西兰大白兔L4，5及L5,6纤维环损伤作为实验组，4只作对照组未行损伤。2、4、8、40周分别行核磁共振及计算机扫描摄影拍片证实椎间盘退变情况同时取椎间盘行精确定量逆转录聚合酶链式反应检测金属蛋白酶(matrix metalloproteinase)MMP-1、MMP-2、MMP-3及它们的特意性抑制剂(tissue inhibitors of metalloproteinases)Timp-1、Timp-2的变化。结果：核磁共振及计算机扫描摄影证实了腰椎纤维环损伤后腰椎逐渐退变并与人类退变结果相似，基因表达情况MMP-1、MMP-2、MMP-3、Timp-1、Timp-2早期均上调，MMP-3、Timp-1在退变的晚期出现下调。结论：纤维环损伤可以成功制作出椎间盘退变的模型。在人类腰椎间盘退变中明显上调的基因在此退变模型椎间盘中被发现同样上调，从而在分子水平证实了动物退变模型与人类的相似性。     

凋亡相关基因产物Fas APO-1蛋白在腰椎间盘组织中的表达及其意义

我们采用免疫组织化学方法对 35例腰椎间盘突出症患者的退变腰椎间盘及 11个 (7例 )正常腰椎间盘组织中ＦａｓＡＰＯ 1蛋白表达进行检测 ,探讨其在腰椎间盘退变及突出症发生、发展中的作用。一、材料与方法1.标本来源 :退变腰椎间盘组织(Ｌ1～ 5)取自本院骨科 1998年 5月～1999年 1月间 35例经手术治疗的腰椎间盘突出症患者 ,其中男 2 0例 ,女 15例 ,年龄 2 0～ 6 9岁 ,平均 40 .0岁。术中取纤维环及髓核组织各 1块 ,标本经病理学检查证实为退变椎间盘。 11个 (7例 )正常椎间盘组织 (Ｌ1～ 5)做为对照 ,标本取自新鲜尸检 ,年龄 2 2～ 5 9岁 …     

Bone morphogenic protein-2 signaling in human disc degeneration and correlation to the Pfirrmann MRI grading system

BACKGROUND CONTEXTBack and neck pain secondary to disc degeneration is a major public health burden. There is a need for therapeutic treatments to restore intervertebral disc (IVD) composition and function.PURPOSETo quantify ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression in IVD specimens collected from patients undergoing surgery for disc degeneration, to correlate ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression in IVD specimens to the 5-level Pfirrmann MRI grading system, and to compare ALK3, BMP-2, pSMAD1/5/8 and MMP-13 expression between cervical and lumbar degenerative disc specimens.STUDY DESIGNAn immunohistochemical study assessing ALK3, BMP-2, pSMAD1/5/8, and MMP-13 expression levels in human control and degenerative IVD specimens.METHODSHuman IVD specimens were collected from surgical patients who underwent discectomy and interbody fusion at our institution between 1/2015 and 8/2017. Each patient underwent MRI prior to surgery. The degree of disc degeneration was measured according to the 5-level Pfirrmann MRI grading system. Patients were categorized into either the 1) control group (Pfirrmann grades I-II) or 2) degenerative group (Pfirrmann grades III-V). Histology slides of the collected IVD specimens were prepared and immunohistochemical staining was performed to assess ALK3, BMP-2, pSMAD1/5/8, and MMP-13 expression levels in the control and degenerative specimens. Expression levels were also correlated to the Pfirrmann criteria. Lastly, the degenerative specimens were stratified according to their vertebral level and expression levels between the degenerative lumbar and cervical discs were compared.RESULTSFifty-two patients were enrolled; however, 2 control and 2 degenerative patients were excluded due to incomplete data sets. Of the remaining 48 patients, there were 12 control and 36 degenerative specimens. Degenerative specimens had increased expression levels of BMP-2 (p=.0006) and pSMAD1/5/8 (p<.0001). Pfirrmann grade 3 (p=.0365) and grade 4 (p=.0008) discs had significantly higher BMP-2 expression as compared to grade 2 discs. Pfirrmann grade 4 discs had higher pSMAD1/5/8 expression as compared to grade 2 discs (p<.0001). There were no differences in ALK3 or MMP-13 expression between the control and degenerative discs (p>.05). Stratifying the degenerative specimens according to their vertebral level showed no significant differences in expression levels between the lumbar and cervical discs (p>.05).CONCLUSIONSBMP-2 and pSMAD1/5/8 signaling activity was significantly upregulated in the human degenerative specimens, while ALK3 and MMP-13 expression were not significantly changed. The expression levels of BMP-2 and pSMAD1/5/8 correlate positively with the degree of disc degeneration measured according to the Pfirrmann MRI grading system.CLINICAL SIGNIFICANCEBMP-SMAD signaling represents a promising therapeutic target to restore IVD composition and function in the setting of disc degeneration.     

MMP-3在退变腰椎间盘组织中的表达及意义

目的研究MMP-3在退变腰椎间盘髓核和纤维环组织中的表达及其临床意义。方法用半定量RT—PCR和免疫组织化学法检测实验组1（30例退变腰椎间盘髓核）、实验组2（30例退变腰椎间盘纤维环）和对照组（10例创伤腰椎间盘髓核）中MMP3mRNA和蛋白表达。结果实验组1MMP-3mRNA和蛋白的表达均显著高于对照组（P均〈0．01）,实验组1与2之间MMP-3mRNA和蛋白的表达没有显著差异（P均〉0．05）。结论MMP-3的表达增加可能参与腰椎间盘退变的进程。     

退变性腰椎侧凸形成和发展的相关因素分析

目的 探讨退变性腰椎侧凸患者椎问盘的不对称指数、腰椎间盘退变程度以及骨密度降低对侧凸角度的影响.方法 采用回顾性研究的方法,选取2002年1月至2010年8月,共96例退变性腰椎侧凸患者为研究对象(侧凸组);2002年1月至2010年8月确诊为腰椎管狭窄症并且资料齐全的患者96例为对照组;两组间性别、年龄、体质量指数匹配.侧凸组:在腰椎正位X线片上测量凸凹侧顶椎间盘及其上下椎间盘的高度和顶椎及其上下椎体的高度,利用Adobe Photoshop 6.0软件,测量MRI图像T2WI顶椎及其上下椎间盘内髓核与脑脊液的相对信号强度.对照组:取2～3、L3-4、L4-5这3个椎间盘为研究对象测定上述指标.应用双能X线吸收法测定两组患者腰椎(L2-4)及股骨颈、股骨粗隆和Ward's三角的T值.结果 侧凸组凸侧椎间盘高度和为(40±7)mm高于凹侧的(28±7)mm(P<0.01),凸侧椎体高度和为(76±12)mm高于凹侧的(72±10)mm(P=0.016):两组之间的椎间盘退变程度差异有统计学差异(P=0.003);两组之间骨密度T值的平均值和骨质疏松的发生率差异有统计学意义(均P<0.01).通过多元线性回归分析结果 显示患者椎间盘的不对称指数、椎间盘的退变程度、骨密度T值影响退变性腰椎侧凸角度.结论 退变性腰椎侧凸常伴有凸凹两侧椎间盘高度以及椎体高度不对称.侧凸角度与椎间盘的不对称指数、椎间盘的退变程度呈正相关,与骨密度值T值呈负相关.

Abstract：

Objectives To investigate the correlation between scoliosis angle and the asymmetric index of degenerative lumbar scoliosis, the degree of intervertebral disc degeneration, decreased bone density. Methods As a retrospectively study, a total of 96 patients with degenerative lumbar scoliosis were retrospectively enrolled from January 2002 to August 2010 as scoliosis group, meanwhile % patients with lumbar spinal stenosis matched in gender, age and body mass index (BMI) were selected as control group.All patients were studied with plain radiographs, MRI and dual energy X-ray absorptiometry at presentation. Radiographic measurements include Cobb angle, the height of the convex and concave side of the apical disc and the contiguous disc superiorly and inferiorly, the height of the convex and concave side of the apical and the contiguous vertebral body superiorly and inferiorly in scoliosis group, the height of L2-3, L3-4, L4-5 discs and the height of L2-4 vertebral body in control group. The average relative signal intensity of lumbar intervertebral disc and cerebrospinal fluid in T2WI sagittal image was measured in apex intervertebral disc and adjacent discs by Adobe Photoshop 6.0 in scoliosis group, which was measured in L2-3, L3-4, L4-5 disc in control group. The bone density of lumbar, femoral neck, trochanter, and Ward's triangle regions were measured with dual-energy X-ray absorptiometry. Results The intervertebral disc height in convex side was greater than the height in the concave side [(40 ± 7) mm vs. (28 ± 7) mm, P < 0. 01] , the vertebral body height in convex side was greater than the height in the concave side [(76 ± 12) mm vs. (72 ± 10) mm, P =0.016] in scoliosis group. There was significant statistically difference in the degenerative degree of intervertebral discs between two groups (P = 0. 003). There was significant statistically difference of the average T-value and the rate of osteoporosis between two groups (P < 0. 01). Multiple linear regression analysis showed that the asymmetric disc index, the degenerative degree of intervertebral disc and osteoporosis were the predominant correlative factors, which affected the development of degenerative lumbar scoliosis. Conclusions Degenerative lumbar scoliosis is always accompanied by the height asymmetry of intervertebral discs and vertebral body from convex and concavity sides. There is positive correlation between the angle of scoliosis and the asymmetric disc index, the degeneration of intervertebral disc, and negative correlation between the angle of scoliosis and the bone density (T-value).     

Herniated and spondylotic intervertebral discs of the human cervical spine: histological and immunohistological findings in 500 en bloc surgical samples. Laboratory investigation

OBJECT: In this paper the authors' goal was to identify histological and immunohistochemical differences between cervical disc hemrniation and spondylosis. METHODS: A total of 500 cervical intervertebral discs were excised from 364 patients: 198 patients with disc herniation and 166 patients with spondylosis. We examined en bloc samples of endplate-ligament-disc complexes. Types of herniation and graded degrees of disc degeneration on MR images were examined histologically and immunohistochemically. RESULTS: The herniated discs showed granulation tissue, newly developed blood vessels, and massive infiltration of CD68-positive macrophages, which surrounded the herniated tissue mainly in the ruptured outer layer of the anulus fibrosus. The vascular invasion was most significant in uncontained (extruded)-type herniated discs. Chondrocytes positive for matrix metalloproteinase (MMP)-3, tumor necrosis factor (TNF)-alpha, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were abundant in both herniated and spondylotic discs. Free nerve fibers, positive for nerve growth factor (NGF), neurofilament 68, growth-associated protein (GAP)-43, and substance P, were strongly apparent in and around the outer layer of uncontained (extruded)-type herniated discs, with enhanced expression of NGF. The authors observed that herniated discs showed more advanced degeneration in the outer layer of the anulus fibrosus around the granulation tissue than spondylotic discs. On the other hand, spondylotic discs showed more advanced degeneration in the cartilaginous endplate and inner layer of the anulus fibrosus than herniated discs. Spondylotic discs also had thicker bony endplates and expressed TNFalpha and MMP-3 more diffusely than herniated discs, especially in the inner layer of the anulus fibrosus. CONCLUSIONS: The authors' results indicate that herniated and spondylotic intervertebral discs undergo different degenerative processes. It is likely that TNFa, MMP-3, bFGF, and VEGF expression is upregulated via the herniated mass in the herniated intervertebral discs, but by nutritional impairment in the spondylotic discs. Macrophage accumulation around newly formed blood vessels in the herniated disc tissues seemed to be regulated by MMP-3 and TNFalpha expression, and both herniated and spondylotic discs exhibited marked neoangiogenesis associated with increased bFGF and VEGF expression. Nerve fibers were associated with NGF overexpression in the outer layer of the anulus fibrosus as well as in endothelial cells of the small blood vessels.     

P物质阳性神经纤维在退变腰椎间盘的分布及意义

目的研究P物质（substance P,SP）阳性神经纤维在腰椎间盘中的出现及分布,以探讨其在椎间盘退变中的潜在作用。方法收集16例患者的21个病变腰椎间盘和来自于新鲜尸体的12个正常对照椎间盘,行组织学检查和SP免疫组织化学染色检查。结果SP阳性神经纤维偶见于正常椎间盘,在病变椎间盘内可见较多的SP免疫反应阳性神经纤维分布,阳性神经纤维数量与腰椎间盘退变程度呈正相关。结论腰椎间盘退变程度和SP阳性神经纤维数量有明显的相关性,SP可能作为神经炎性介质加速了腰椎间盘的退变。