肥胖与腰椎间盘退变的关联性临床研究

目的探讨肥胖与腰椎间盘退变的关联性。方法筛选符合既定相关标准的研究对象150例,分为病例观察组60例、病例对照组60例、健康组30例,均进行MRI检查,再根据Pfirrmann分级方法判断腰椎间盘退变的程度,然后运用统计软件对所得资料进行分析。结果病例观察组及病例对照组腰椎间盘退变为Ⅴ级与健康组腰椎间盘退变为Ⅴ级所占比例比较差异有统计学意义(P<0.05);病例对照组中,肥胖者腰椎间盘退变为Ⅳ～Ⅴ级所占比例与超重及正常体重者间腰椎间盘退变为Ⅳ～Ⅴ级所占比例相比较差异有统计学意义(P<0.05),健康组中,肥胖者的腰椎间盘退变程度也比超重及正常体重者严重。结论体质指数越大,腰椎间盘退变的程度越高,肥胖可能是引发腰椎间盘退变的危险因素之一。     

CircRNA expression profile and potential role of hsa_circ_0040039 in intervertebral disc degeneration

Purpose:Circular RNAs (circRNAs) play an critical role in the pathological processes associated with IDD. However, the potential roles of circRNAs in IDD remain largely unclear. Here, we identify the circRNAs expression profiles and elucidate the potential role of candidate circRNAs in the pathogenesis of intervertebral disc degeneration (IDD) through microarray data and bioinformatics analyses.Methods:We obtained the datasets of microarrays ({"type":"entrez-geo","attrs":{"text":"GSE67566","term_id":"67566"}}GSE67566 and {"type":"entrez-geo","attrs":{"text":"GSE116726","term_id":"116726"}}GSE116726) from the Gene Expression Omnibus database. The differentially expressed circRNAs and miRNAs were identified using the Limma R package. The target miRNAs and target genes of the candidate circRNAs were predicted using an online tool. Functional enrichment analyses of the target genes were performed using the clusterProfiler R package. A protein-protein interaction (PPI) network was constructed using STRING.Results:A total of 104 differentially expressed circRNAs were identified between the IDD and the control groups, including 41 upregulated circRNAs and 63 downregulated circRNAs (cutoff criteria (|log₂ fold change| > 2, P < .05)). Hsa_circ_0040039, which was the most upregulated circRNA (log₂ fold change = 2.95), was selected for further analysis. The regulatory circRNA-miRNA-mRNA network comprised hsa_circ_0040039, 2 target miRNAs (hsa-miR-424-5p and hsa-miR-15b-5p), and 77 target genes. Functional enrichment analysis showed that the 77 promising target genes are mainly enriched in the ubiquitin proteasome system and Wnt signaling pathway. Further, the PPI network showed that the top 3 hub genes are BRTC, SIAH1, and UBE2V1.Conclusions:A total of 104 differentially expressed circRNAs were identified between the IDD and control groups. Hsa_circ_0040039 may serve as a sponge of hsa-miR-424-5p and hsa-miR-15b-5p, to regulate the expression of downstream genes (such as BRTC, SIAH1, and UBE2V1); thus, it may be involved in IDD-associated pathological processes via the Wnt/β-catenin signaling pathway. Further studies are required to confirm the potential roles of hsa_circ_0040039 in IDD.     

糖尿病大鼠椎间盘退变的实验观察

目的 观察糖尿病大鼠椎间盘的退变情况,探讨糖尿病引起椎间盘退变的可能机制.方法 30只雄性SD大鼠随机分为实验组和对照组,每组15只.实验组采用腹腔单次注射链脲佐菌素溶液(STZ)50 mg/kg制成糖尿病模型,正常对照组仅注射等量的枸橼酸缓冲液.分别在建模成功后4、8、12 w取椎间盘组织,HE染色观察椎间盘组织病理变化;透射电镜观察髓核细胞及胶原变化;用TUNEL法显示髓核组织细胞凋亡情况,测定凋亡率. 结果 实验组大鼠注射STZ后,空腹血糖均大于13.8 mmol/L[(20.27±2.600)mmol/L].在不同的阶段,实验组椎间盘的退变程度均较重,髓核细胞凋亡率较高.结论 糖尿病是影响椎间盘退变的因素之一.     

MMP-3在大鼠突出椎间盘组织及血液中的表达与意义

目的检测大鼠突出椎间盘组织及血液中基质金属蛋白酶3(MMP-3)含量的变化,探讨MMP-3保守治疗椎间盘突出症的可行性。方法随机将40只3月龄SD大鼠,分为正常对照组、阴性对照组、实验组1及实验组2,每组10只。造模45 d后,用ELISA法检测椎间盘突出组织及血液中MMP-3含量。结果(1)与正常对照组相比,实验组1、2的椎间盘组织及血液MMP-3浓度上升,差异有统计学意义(P0.05)。(2)实验组1椎间盘组织及血液MMP-3浓度较实验组2明显升高,差异有统计学意义(P0.05)。(3)实验组1、2椎间盘组织中MMP-3浓度的变化与血液中MMP-3相关系数分别为0.835和0.924,均P0.05。结论椎间盘突出再吸收过程中,血液中MMP-3含量上升,且局部椎间盘组织与血液中MMP-3的含量高度正相关,临床上运用MMP-3局部注射非手术方法治疗椎间盘突出症的设想可能行不通。     

cDNA cloning and expression of a metalloproteinase inhibitor related to tissue inhibitor of metalloproteinases.

The purification and characterization of a metalloproteinase inhibitor (MI) from bovine aortic endothelial cells, and the demonstration that it is related to, but distinct from, tissue inhibitor of metalloproteinase (TIMP), have previously been reported [De Clerck, Y. A., Yean, T.-D., Ratzkin, B. J., Lu, H.S. & Langley, K. E. (1989) J. Biol. Chem. 264, 17445-17453]. The cDNA cloning of the bovine MI and its human homolog is now reported. The bovine cDNA cloning used probes designed on the basis of NH2-terminal amino acid sequence of bovine MI. The human cDNA cloning in turn used probes representing parts of the bovine cDNA nucleotide sequence. Both cDNAs encode leader sequences of 26 amino acids and mature protein sequences of 194 amino acids. The amino acid sequences of the mature proteins are 94% identical. The human MI cDNA was expressed in Escherichia coli, and a preparation containing anticollagenase activity was recovered. The amino acid sequence of mature human MI is 38% identical to the sequence for human TIMP, and the 12 cysteines in MI and TIMP are aligned almost identically. Thus MI and TIMP comprise an inhibitor family.     

Human lumbar apophyseal joint damage and intervertebral disc degeneration.

OBJECTIVES--To record the extent and location of lumbar apophyseal cartilage damage, and to ascertain if the extent of damage is correlated with the grade of disc degeneration, age, or both. METHODS--The extent and location of fibrillated areas of the apophyseal cartilage of the joint surfaces of 29 lumbar motion segments were examined using computer aided image processing of Indian ink stained areas, and degeneration of the associated intervertebral discs graded using the method of Nachemson. RESULTS--It was found that these joints showed a greater extent and prevalence of cartilage fibrillation than the knee, hip or ankle, with significant damage in specimens younger than 30 years. Damage was predominantly located peripherally, superiorly, and posteriorly in the concave superior apophyseal surfaces, and was predominantly peripheral and posterior in the inferior surfaces, with a tendency to be located inferiorly. There was a weak correlation between apophyseal joint damage and the intervertebral disc degenerative grade, but this was inconclusive, as both increased with age. CONCLUSIONS--The pattern of damage exhibited by superior joint surfaces is most probably caused by tension on collagenous joint capsule fibres which insert into the surfaces posteriorly, so producing an area of fibrocartilage unsuited to loadbearing. Tension on such fibres would be greatest during spinal flexion. The pattern of damage of the inferior surfaces lends some support to the hypothesis that their apices impact the laminae of the lumbar vertebra inferior to them, consequent upon the degeneration and narrowing of the associated intervertebral disc. The predominantly peripheral location of fibrillation of both superior and inferior surfaces may be associated with inadequate mechanical conditioning of marginal joint areas. Disc degeneration cannot be the initial cause of apophyseal fibrillation in most specimens. The study indicates a need for regular spinal exercise, starting at a young age.     

Proinflammatory cytokine expression profile in degenerated and herniated human intervertebral disc tissues

Objective

Prior reports document macrophage and lymphocyte infiltration with proinflammatory cytokine expression in pathologic intervertebral disc (IVD) tissues. Nevertheless, the role of the Th17 lymphocyte lineage in mediating disc disease remains uninvestigated. We undertook this study to evaluate the immunophenotype of pathologic IVD specimens, including interleukin‐17 (IL‐17) expression, from surgically obtained IVD tissue and from nondegenerated autopsy control tissue.

Methods

Surgical IVD tissues were procured from patients with degenerative disc disease (n = 25) or herniated IVDs (n = 12); nondegenerated autopsy control tissue was also obtained (n = 8) from the anulus fibrosus and nucleus pulposus regions. Immunohistochemistry was performed for cell surface antigens (CD68 for macrophages, CD4 for lymphocytes) and various cytokines, with differences in cellularity and target immunoreactivity scores analyzed between surgical tissue groups and between autopsy control tissue regions.

Results

Immunoreactivity for IL‐4, IL‐6, IL‐12, and interferon‐γ (IFNγ) was modest in surgical IVD tissue, although expression was higher in herniated IVD samples and virtually nonexistent in control samples. The Th17 lymphocyte product IL‐17 was present in >70% of surgical tissue fields, and among control samples was detected rarely in anulus fibrosus regions and modestly in nucleus pulposus regions. Macrophages were prevalent in surgical tissues, particularly herniated IVD samples, and lymphocytes were expectedly scarce. Control tissue revealed lesser infiltration by macrophages and a near absence of lymphocytes.

Conclusion

Greater IFNγ positivity, macrophage presence, and cellularity in herniated IVDs suggests a pattern of Th1 lymphocyte activation in this pathology. Remarkable pathologic IVD tissue expression of IL‐17 is a novel finding that contrasts markedly with low levels of IL‐17 in autopsy control tissue. These findings suggest involvement of Th17 lymphocytes in the pathomechanism of disc degeneration.

     

扩张型心肌病心肌组织中分离整合素金属蛋白酶及其抑制剂的表达改变

背景:分离整合素金属蛋白酶(ADAMs)可能通过改变细胞表面的基质受体(整合素)、激活生物分子而参与心脏结构的重塑。我们在不同模式的重塑心肌组织中比较了ADAMs、其内源性抑制剂TIMP-3和整合素的表达差异。方法和结果:心肌组织取材于扩张型心肌病(n=20例)、梗阻性肥厚型心肌病(n=5例)和心功能正常的供体(n=7例)。匹配的样本(n=10例)来源于左室辅助装置植入之前和心脏移植时。通过免疫印迹法检测 ADAM10、ADAM12、ADAM15和ADAM17、TIMP-3以及β1D整合素受体、β整合素受体的表达。整合素脱落的测量是通过计算整合素裂解产物和完整整合素的比例而得到。扩张型心肌病中ADAM10、ADAM15表达增高而TIMP-3表达降低。β1D整合素裂解产物比例升高,提示受体脱落。ADAM10、ADAM15表达与裂解产物比例相关。共聚焦显微镜观察到ADAM10与β1D整合素的分布部位一致。ADAM10的表达和室腔扩张、收缩功能不全等临床指标相关。血流动力学改善减少ADAM10和ADAM12的表达,却上调β1D整合素的表达。在梗阻性肥厚型心肌病中,ADAM12和β1D整合素的表达均增加。ADAM17在扩张型心肌病和梗阻性肥厚型心肌病中均见表达上调。结论:ADAMs在人心肌组织的表达差异反映了不同的心肌重塑模式,ADAM10和ADAM15 可能通过引起整合素脱落而减少细胞基质的相互作用参与心脏扩张。因此,以ADAMs为靶标,通过基因或细胞治疗恢复缺乏的TIMP-3,可能阻止扩张型心肌病的进行性心腔扩张。     

Synovial membrane expression of matrix metalloproteinases and tissue inhibitor 1 in juvenile idiopathic arthritides

OBJECTIVE: Matrix metalloproteinases (MMP) are a large family of proteolytic enzymes involved in the remodeling of extracellular matrix during tissue resorption. We investigated synovial tissue expression of the main proteolytic enzymes (MMP-1, MMP-3, and MMP-13) and tissue inhibitor of metalloprotease 1 (TIMP-1) in juvenile idiopathic arthritides (JIA). METHODS: Expression of MMP-1, MMP-3, MMP-13, and TIMP-1 was studied by immunohistochemical analysis of synovial tissues, obtained at synoviectomy or arthroplasty from 9 patients with JIA, and was correlated with mononuclear cell infiltration into the lining layer. RESULTS: MMP-1 and MMP-3 were abundantly expressed in the lining layer, showing a high degree of correlation with macrophage infiltration (CD68+ cells). MMP-13 showed a lower degree of expression, with tissue distribution almost restricted to the sublining regions. TIMP-1 tissue distribution was similar to that observed for MMP-1 and -3, although with a definitely lower number of positive cells. CONCLUSION: The expression of MMP-1 and MMP-3 in the synovium of patients with IA was clearly correlated with the degree of inflammation. This indicates the possible role of MMP in the pathogenesis of synovitis in this group of pediatric idiopathic arthritides. Inadequate expression of tissue inhibitors may represent a crucial event for the development and perpetuation of tissue damage.     

Experimental intervertebral disc degeneration. morphologic and proteoglycan changes over time

An animal model of intervertebral disc degeneration produced by surgical ventral disc herniation in the rabbit is described. Histologic studies showed proliferation of cells on the inner third of the annular wound, with metaplasia into fibrocartilage in the first 2 weeks following injury. Progressive fibrocartilaginous change occurred, reproducing the morphologic changes of disc degeneration over the first 6 weeks involving nearly the entire disc. Proteoglycans (total and newly synthesized) were studied qualitatively and quantitatively for periods of 1 to 200 days after herniation. There were two periods of time during the early course of degeneration when the ability of the proteoglycans to aggregate by interaction with hyaluronic acid was recovered, but this decreases progressively after 6–7 weeks. There was an immediate loss of water content from the injured disc which was restored only transiently during the first 2 days after herniation. Thereafter the water content progressively decreased. The uronic acid content of the disc changed in parallel with the changes in water content. The hyaluronic acid content decreased rapidly after herniation. However, the size of the proteoglycan monomers did not change with degeneration. The biochemical and morphologic changes are correlated, and an early repair mechanism is postulated to exist after injury.     

组织金属蛋白酶抑制物在老年大鼠肾小管间质病理损害中的动态变化

目的研究组织金属蛋白酶抑制物(TIMPs)及基质金属蛋白酶(MMPs)在不同鼠龄单侧输尿管梗阻(UUO)大鼠肾小管间质中的表达变化,探讨其在老年大鼠肾小管间质损害中的作用. 方法选用3月龄(青年)、26月龄(老年)大鼠制备UUO模型,采用病理及免疫组化等方法动态观察不同年龄大鼠UUO术后3、7、14d梗阻侧肾脏的组织学变化和TIMP-1、TIMP-2 、MMP-2、MMP-9等在梗阻肾小管间质中的表达情况. 结果 TIMP-1、TIMP-2及MMP-2、MMP-9主要表达于肾小管上皮细胞和间质细胞.在对照组中,TIMP-1、TIMP-2仅微弱表达于青年鼠肾脏中,而在老年大鼠肾脏中其表达显著增加(TIMP-1 0.84±0.23 vs 2.17±0.85,P<0.01; TIMP-2 0.93±0.12 vs 2.31±0.87,P<0.01),MMP-2、MMP-9在正常青年鼠与老年鼠肾脏中均有表达,二者之间差异无显著性(P＞0.05).与对照组相比,3、26月龄大鼠梗阻侧肾脏的TIMP-1、TIMP-2及MMP-2、MMP-9在术后各时间点的表达均显著增高(P<0.01),肾小管间质纤维化面积随UUO术后时间的延长而增加,且TIMP-1、TIMP-2的表达与肾小管间质纤维化面积呈正相关(TIMP-1,r=0.816,P<0.01; TIMP-2, r=0.851, P<0.01).与3月龄UUO大鼠比较,26月龄UUO大鼠TIMP-1、TIMP-2在肾小管间质中各时间点的表达均显著增高(P<0.01),肾小管间质纤维化面积在各时间点也明显增加(P<0.01). 结论 TIMPs的高表达所致的MMPs/TIMPs比例失调可能促进老年大鼠肾小管间质损害的进程.     

Neck and back pain and intervertebral disc degeneration: role of occupational factors

Back pain is a near-universal human experience at some time during life, and neck pain is also common. The overwhelming majority of low back and cervical pain is considered to be due to unspecified mechanical factors or disc degeneration, which is a common with ageing and, hence, in people of working age. Back pain and disc disease appear to have significant heritability, based upon twin studies, but environmental factors also contribute - including physical occupational activities in some studies - although the strength of this association remains uncertain. This article examines the contribution of genetic and environmental factors to back pain and disc disease, with a specific focus on occupational exposures.     

基质金属蛋白酶抑制剂和肝纤维化关系的研究进展

基质金属蛋白酶抑制剂（TIMPs）是基质金属蛋白酶（MMPs）的特异性抑制剂,两者比例失常将导致细胞外基质合成和降解失去平衡,促进肝纤维化的形成。TIMPs表达的异常在肝纤维化形成过程中具有非常重要的作用,本文对TIMPs在肝纤维化形成中的变化及其对肝纤维化诊断、治疗等方面的作用作一综述。     

骨密度与颈椎间盘退变关系的实验研究

目的 探讨骨密度与颈椎间盘退变的相互关系. 方法 利用14月龄雌性大鼠,采用去势造成骨质疏松(A组)、破坏鼠颈后部软组织结构造成颈椎间盘退变(B组)、及两者合并(C组)等3种动物模型,通过观察各组鼠骨密度(BMD)、颈椎间盘退变的形态学分级变化,与正常组(D组)对比,进行相关性分析. 结果 B组中,颈椎间盘退变程度与颈椎BMD呈正相关(P<0.01).在C组中,颈椎间盘退变程度与颈椎BMD呈正相关(P<0.05),与腰椎BMD呈负相关(P<0.01),在其余各组中,也有这种趋势,但没有统计学意义(P0.05).各组中,全身BMD与颈椎间盘退变程度之间的关系,没有显著意义(P0.05). 结论 颈椎间盘退变与颈椎骨密度呈正相关,与腰椎骨密度呈负相关,与全身骨密度没有关系.