压力可控型椎间盘退变模型的建立及相关研究

[目的]建立压力可控型椎间盘退变模型并进行相关研究.[方法]80只成年大鼠随机分为A、B、C、D 4组,分别模拟人类在站立(A组)、坐位直立(B组)、坐位前屈(C组)3种状态下椎间盘内的压力情况,应用恒定加压装置对第9、10尾椎加压,D组为对照组(一般人椎间盘面积为1 250 mm3,大白鼠为2.8 mm2,人在站立位时椎间盘内压力为500 N,坐位为750 N,坐位前屈为1 350 N,换算成大白鼠椎间盘的压力分别是1.12 N,1.68 N,3.08 N).A、B、C组每日加压4 h,对照组仅在尾椎穿针,不加压;4组分别在7、14 d后取标本进行组织学观察,应用免疫组化和阿利新蓝分析测量胶原纤维(Ⅱ型)和蛋白多糖含量,观察椎间盘退变情况.[结果](1)A组、D组类似,椎间盘组织形态、基质胶原和PG含量和成分无明显变化;(2)C组椎间盘发生软骨细胞变性、坏死,髓核皱缩,纤维环胶原纤维变性、排列紊乱,软骨终板钙化、断裂,胶原纤维排列紊乱等组织形态学改变.髓核和软骨终板中PG总量和Ⅱ型胶原表达下降(P<0.05);(3)B组14 d时形态学未观察到软骨细胞坏死等退变表现,但Ⅱ型胶原以及PG含量比对照组明显减少(P<0.05),退变程度较C组轻.[结论]应用恒定加压装置模拟人类脊柱坐位前屈压力加压大鼠尾椎可迅速建立大鼠椎间盘退变模型.     

可控轴向压力致兔腰椎间盘退变模型的建立及评价

[目的]建立可控轴向压力诱发的椎间盘退变动物模型并考察其特性.[方法]取4个月龄日本大白兔40只,将其随机分为5组(每组8只).采用基于横穿椎体的克氏针的自制加压装置,分别对2～4组动物腰椎间盘施加10 kg压力1、2、4周及8周,第1组作为假手术对照组.Thompson分级法及兔腰椎间盘磁共振(MRI)评价退变程度,HE染色及Ⅰ、Ⅱ型胶原免疫组化染色考察其组织学改变.另外构建纤维环损伤致兔腰椎间盘退变模型,并对标本行HE染色以供比较.[结果]①Thompson分级:第2组7只均为Ⅱ级,第3组6只Ⅲ级,第4组5只为Ⅳ级,第5组4只动物退变程度达到V级.②MRI评分显示,第2组椎间盘以轻度退变为主;第3组椎间盘主要为中度退变.第4组、第5组则以重度退变为主.③HE染色显示,该模型椎间盘各区域的退变情况较纤维环损伤模型更为均匀.④免疫组化染色显示,退变过程中椎间盘基质成分退化的趋势与人体相符.[结论]可控轴向压力致兔腰椎间盘退变模型可以高效诱发椎间盘退变,尤其在轻、中度椎间盘退变方面具有优势.该模型可供深入研究,并用于椎间盘退变的治疗研究.     

纤维环穿刺法建立兔椎间盘退变模型

[目的]通过纤维环穿刺法建立兔椎间盘退变模型,分析其特点.[方法]将16只新西兰大白兔随机分为2组.经腹膜外入路暴露X2、3、L3、4椎间隙.A组为纤维环穿刺组,采用16号针头穿刺,B组为假手术对照组.术后2,4,6,8周通过MRI和组织病理学检查观察髓核变性及组织病理情况.[结果)纤维环穿刺组髓核信号强度在术后呈现逐渐降低趋势,髓核面积逐渐缩小,椎间隙高度也逐步下降,从术后4周开始两组差异有统计学意义(p＜0.O5 ).随着时间的进展,纤维环穿刺组髓核内细胞含量逐渐减少.[结论]纤维环穿刺法可成功建立椎间盘退变模型,比较真实地模拟了人类椎间盘损伤后的退变过程.     

椎间盘退变模型的建立及其历史和现状

腰腿痛是影响人类健康的常见病和多发病。研究表明约２３％的腰腿痛患者其病因与椎间盘的退行性改变有关［１］。但椎间盘退变的确切病因及病理生理机制目前仍不十分清楚。为深入探讨椎间盘退变的病理过程,有必要建立类似人类椎间盘退变的动物实验模型,对椎间盘退变的诱发因素,早期发生的形态学、生物化学及其它方面的变化进行较为详尽的研究。 一、动物模型的要求及动物选择 建立的动物模型要求与人类的椎间盘退变具有相似性和可比拟性。应包括以下几个方面：（１）能再现椎间盘退行性病变的客观规律;（２）模型重复性好;（３）所选动…     

应力诱导椎间盘退变的研究进展

腰背痛是一种常见的脊柱退行性疾病,目前已经发展成为严重的社会问题。近期研究表明,椎间盘退变（intervertebral disc degeneration, IDD）是导致腰背痛的首要原因,抑制IDD是防治腰背痛的重要靶点。导致IDD的原因包括老龄、遗传、机械负荷、营养缺乏等。近年来,越来越多的研究报道了机械应力（mechanical stress, MS）导致的IDD,MS诱导IDD的分子机制复杂多变,包括调控凋亡、焦亡、铁死亡、衰老、自噬、氧化应激、内质网应激、炎性反应和ECM降解等,但其确切机制尚不明确。本文拟对MS诱导IDD的机制进行综述,为临床预防和治疗腰背痛提供理论参考。     

椎间盘退变机制及动物模型建立的研究进展

椎间盘退变常会引起腰背痛.长期以来人们对椎间盘退变机制进行了大量的探索与研究,目前普遍认为椎间盘退变是一个多因素诱导的(遗传因素、环境因素、年龄因素)、多种因子(生长因子、炎性因子)参与的不可逆的过程,其确切机制尚不明确.模型的建立可与退变机制的研究相互论证,益于对退变机制的探索.     

椎间盘退变模型研究进展

椎间盘退变的生物学机制尚不明确,构建椎间盘退变模型是研究基础和关键。近年来国内外报道的新型椎间盘退变模型分为体外和体内培养模型两大类。体外模型包括椎间盘细胞模型和椎间盘组织块模型,多用于细胞生物学行为方面的研究,适用面较广,但培养要求较高,模拟体内环境有局限。体内模型有机械力学模型、外伤模型、酶化学模型、缺血模型、吸烟模型等,多用于采取特定干预措施的椎间盘退变模型的研究,适用面较窄,但干预技术较易实现。该文就近年来新型椎间盘退变模型的研究进展及各种模型的优缺点作一综述。     

椎间盘退变模型研究进展

椎间盘退变的生物学机制尚不明确,构建椎闯盘退变模型是研究基础和关键.近年来国内外报道的新型椎间盘退变模型分为体外和体内培养模型两大类.体外模型包括椎间盘细胞模型和椎间盘组织块模型,多用于细胞生物学行为方面的研究,适用面较广,但培养要求较高,模拟体内环境有局限.体内模型有机械力学模型、外伤模型、酶化学模型、缺血模型、吸烟模型等,多用于采取特定干预措施的椎间盘退变模型的研究,适用面较窄,但干预技术较易实现.该文就近年来新型椎间盘退变模型的研究进展及各种模型的优缺点作一综述.     

渗透压负荷对兔椎间盘器官培养模型的影响

目的:建立一种可用于体外研究椎间盘退变的椎间盘器官培养模型,探讨渗透压负荷对模型椎间盘细胞活力和代谢的影响.方法:4～6月龄新西兰大白兔10只,均分为两组,处死后立即手术切取胸腰段椎间盘,每只9个,分别在等渗(300mOsm/kg,等渗组)或高渗(410mOsm/kg,高渗组)培养基中进行整体器官培养,在培养前和培养后第7、14、21、28天,利用Mitotracker Green荧光探针、组织化学和生物化学方法评估两组椎间盘髓核细胞的活力、结构的完整性以及蛋白多糖含量的变化.结果:取材后培养前椎间盘髓核细胞的荧光强度为11503±402,在体外培养过程中,高渗组第14天荧光强度为9202±907,与培养前比较差异有显著性意义(P＜0.05),第7、21和28天分别为10504±710、10860±711、10713±953,与培养前相比较无显著性差异(P＞0.05);等渗组第7、14、21、28天分别为11350±351、11207±385、10914±300、10862±229,与培养前比较无显著性差异(P＞0.05);两组在第7、21和28天时无显著性意义(P＞0.05).在培养过程中,两组椎间盘髓核和纤维环的组织结构能够基本保持完整,髓核蛋白多糖含量在培养第7天高渗组和等渗组分别为3.33±0.28m/100mg和2.83±0.25m/100mg,均较培养前(5.03±0.37m/100mg)明显降低(P＜0.01),第14、21、28天时与第7天相比下降不明显(P＞0.05).两组相同时间点的差异无显著性意义(P＞0.05).结论:椎间盘器官培养模型可以在等渗或高渗环境中有效维持兔椎间盘结构的完整性和髓核细胞的活力至少4周.     

兔椎间盘器官与脊柱运动节段离体培养条件下髓核组织的变化

目的:比较离体培养的兔椎间盘器官及脊柱运动节段两种模型椎间盘髓核组织的变化.方法:将21只新西兰白兔随机分为器官组8只,节段组13只,处死后在无菌条件下分别取出椎间盘器官和脊柱运动节段各50个,在高渗培养基中进行整体培养(410 mOsm/kg),于培养前及培养后第3、7、14、21天,两组各取10个椎间盘分别进行HE染色、II型胶原免疫组化、蛋白多糖含量和髓核细胞活力测定.结果:培养21 d器官组与培养14 d节段组HE染色示椎间盘组织结构基本保持完整,21 d节段组椎间盘组织形态学破坏;21 d器官组与14 d节段组Ⅱ型胶原免疫组化染色强度差异无统计学意义(P>0.05),21 d节段组染色变浅,与之前各时间点及器官组相比差异有统计学意义(P<0.05);蛋白多糖PAS/AB染色7 d内两组强度无降低,14 d两组强度均有所减弱,21 d节段组染色强度进一步减弱,改变比器官组更为明显;髓核细胞荧光检测两组7 d时强度较培养前变化不明显(P>0.05),21 d器官组与14 d节段组强度略有降低,但与之前时间点比较差异无统计学意义(P>0.05),21 d节段组髓核细胞荧光强度减弱,细胞活性降低,与之前各时间点及器官组比较差异明显(P<0.05).结论:14d内脊柱运动节段可作为研究生物力学对椎间盘影响的离体实验模型.     

Characterization of an in vitro intervertebral disc organ culture system

Intervertebral disc organ culture has the capacity to control mechanical and chemical boundary conditions while keeping the tissue largely intact, and allowing interventions that would be impossible or unethical on animal studies. Recent studies on ex vivo organ culture has mostly involved small animals, or been limited to development and validation studies. In this study, bovine caudal discs were used. The large animal model design ensures that sufficient tissue is available for measurement of multiple dependent variables on the same disc, and a similar aspect ratio, diffusion distance, composition and rate of proteoglycan synthesis to human lumbar discs. The first goal of this study was to refine a set of dependent variables capable of characterizing the response of the intervertebral disc to culturing and to develop a technique to measure cell viability in all three regions of the disc. The second goal was to use these variables to compare static and diurnal loading as a method of maintaining intervertebral disc structure, composition, and cell metabolism similar to the in vivo state. Static (0.2 MPa) and diurnal loading (0.1 and 0.3 MPa alternating at 12 h intervals) were applied and intervertebral discs were examined after 4 or 8 days with dependent variables including changes in geometry (disc height and diameter), composition (tissue water content, tissue proteoglycan content and proteoglycan content lost to the culture media), cell viability and metabolism (proteoglycan synthesis). Results indicate that there was a decrease in disc height and water content after culture regardless of culture duration or loading condition. Cell viability significantly decreased with culture duration in the inner annulus and nucleus; however, a significant reduction in cell viability for the diurnal versus static loading condition was only observed after 8 days in the nucleus region. No significant differences were seen in viability of the outer annulus region with time, or in any loading groups. We conclude that our system is capable of keeping bovine caudal discs alive for at least 8 days without significant changes in GAG content, or cell metabolism, and that static loading was slightly better able to maintain cell viability than diurnal loading. This system offers promise for the future studies on large intervertebral discs requiring measurements of multiple mechanical and biological dependent variables on the same tissue.     

大鼠尾椎椎间盘退变静态压力模型的构建

目的采用静态压力构建稳定的大鼠尾椎椎间盘退变模型。方法将40只12周龄大鼠随机分为静态压力组和针刺组,每组20只。静态压力组:在尾椎C_(4~7)椎体上安装静态压力环形外固定支架,压缩的4根弹簧施加的压强均为10 kPa,维持8周。针刺组:采用16 G针头刺入尾椎C_4/C_5/C_6/C_7椎间盘,限制损伤深度5 mm、朝向椎间盘中心,损伤后留置10 s。分别于术后8周进行组织病理学切片观察,采用秩和检验比较2组Thompson椎间盘退变病理分级。结果对2组大鼠尾椎共120个椎间盘髓核组织标本进行病理分级,统计结果显示静态压力组退变级别明显高于针刺组,差异有统计学意义(P0.05)。结论采用静态压力构建的椎间盘退变模型稳定可靠,较传统针刺模型具有优势,可为临床椎间盘退变研究提供可靠的动物模型。     

软骨调节素在成人退变椎间盘中的表达

目的 利用分子生物学及免疫组化方法研究软骨调节素(ChM-Ⅰ)在成人退变椎间盘细胞及椎间盘组织中的表达情况.方法 2009年3月至4月取3例因腰椎间盘退变性疾病而需行后路椎间融合患者的椎间盘组织分别进行髓核及纤维环细胞培养.取部分原代细胞利用RT-PCR和Western blot研究ChM-Ⅰ mRNA和蛋白在成人退变椎间盘细胞中的表达情况.利用real-time PCR和Western blot研究不同浓度碱性成纤维细胞生长因子(bFGF)对髓核细胞和纤维环细胞ChM-Ⅰ mRNA和蛋白表达的影响.收集2008年10月至2009年10月因腰椎间盘退变性疾病而行手术切除的椎间盘组织标本26例,根据MRI表现退变程度为Ⅲ～V级,作为退变组.收集同期因脊柱肿瘤行手术治疗时切除的椎间盘标本6例,退变程度Ⅰ级,作为对照组.利用免疫组化方法研究ChM-Ⅰ在不同退变程度椎间盘组织中的表达情况.结果 RT-PCR、Western blot显示ChM-Ⅰ在椎间盘髓核细胞及纤维环细胞中均有表达.bFGF可抑制ChM-Ⅰ在髓核及纤维环细胞中的表达,且呈剂量依赖性(P＜0.05).ChM-Ⅰ在对照组椎间盘组织中表达量很低,阳性细胞率为0.12±0.03,而在椎间盘发生退变后其表达量明显升高,退变组与对照组相比差异有统计学意义(P＜0.05).结论 髓核细胞及纤维环细胞可表达ChM-Ⅰ,bFGF可明显抑制ChM-Ⅰ mRNA及蛋白的表达.椎间盘发生退变后ChM-Ⅰ表达明显升高,提示其可能在椎间盘退变的病理过程中发挥一定的作用.

Abstract：

Objectives To investigate the expression of chondromodulin-1(ChM-Ⅰ)in human adult degenerative intervertebral disc(IVD)cells and the relationship between ChM-Ⅰ expression and disc degeneration.Methods Three degenerated disc specimens obtained from patients in the treatment of disc degenerative disease from March to April 2009 were used for cell culture.ChM-Ⅰ expression in ⅣD cells was examined by RT-PCR and Western blot.The effect of basic fibroblast growth factor(bFGF)on the expression of ChM-Ⅰ was assessed by real-time PCR and Western blot.From October 2008 to October 2009,26 human ⅣD tissues were obtained from patients in the surgical treatment of disc degenerative disease at different stage of degeneration according to MRI.Six IVD tissues removed from patients with metastatic spinal tumor were used as normal control.The expression of ChM-Ⅰ determined by immunohistochemical analysis was correlated with MRI degeneration grade.Results RT-PCR and Western blot examination showed that ChM-Ⅰ was expressed in both adult degenerative anulus fibrosus and nucleus pulposus cells.The mRNA and protein expression of ChM-Ⅰ were both down-regulated by administration of bFGF with dose-dependent way(P＜0.05).Immunohistochemical analysis showed the percent of ChM-Ⅰ immunopositive cells in the control group was 0.12 ± 0.03,and the number increased significantly in the advanced degeneration group(P＜ 0.05).Conclusions The current results demonstrate that ⅣD cells express ChM-Ⅰ.Administration of bFGF down-regulates the expression of ChM- Ⅰ.The expression of ChM-Ⅰ is correlated with the degree of ⅣD degeneration which means it may involve in the process of ⅣD degeneration.     

Development of an intact intervertebral disc organ culture system in which degeneration can be induced as a prelude to studying repair potential

The present work describes a novel bovine disc organ culture system with long-term maintenance of cell viability, in which degenerative changes can be induced as a prelude to studying repair. Discs were isolated with three different techniques: without endplates (NEP), with bony endplates (BEP) and with intact cartilage endplates (CEP). Swelling, deformation, and cell viability were evaluated in unloaded cultures. Degeneration was induced by a single trypsin injection into the center of the disc and the effect on cell viability and matrix degradation was followed. Trypsin-treated discs were exposed to TGFβ to evaluate the potential to study repair in this system. NEP isolated discs showed >75% maintained cell viability for up to 10 days but were severely deformed, BEP discs on the other hand maintained morphology but failed to retain cell viability having only 27% viable cells after 10 days. In CEP discs, both cell viability and morphology were maintained for at least 4 weeks where >75% of the cells were still viable. To mimic proteoglycan loss during disc degeneration, a single trypsin injection was administered to the center of the disc. This resulted in 60% loss of aggrecan, after 7 days, without affecting cell viability. When TGFβ was injected to validate that the system can be used to study a repair response following injection of a bio-active substance, proteoglycan synthesis nearly doubled compared to baseline synthesis. Trypsin-treated bovine CEP discs therefore provide a model system for studying repair of the degenerate disc, as morphology, cell viability and responsiveness to bio-active substances were maintained.     

基质细胞衍生因子-1在人正常和退变椎间盘中的表达、分布及意义

目的:比较人正常和退变椎间盘髓核中央区、髓核与纤维环交界区、纤维环外层、纤维环内层及软骨终板5个相邻部位的基质细胞衍生因子-1 (SDF-1)的表达情况,探讨退变椎间盘SDF-1的表达在上述五个相邻部位是否具有差异.方法:获取20例腰椎间盘突出症患者的椎间盘(L4/5,病理证实为退变椎间盘组织),并纳入退变组;7例急性腰椎爆裂骨折患者的椎间盘(L3/4)及3例脊柱侧凸患者的椎间盘(L2/3,病理证实为正常椎间盘组织),纳入正常组.HE染色观察两组椎间盘的组织学结构,免疫组织化学、Western-blot、rt-PCR检测椎间盘髓核中央区、髓核与纤维环交界区、纤维环内层、纤维环外层及软骨终板5个部位的SDF-1表达情况及分布,并将退变椎间盘上述5个部位的SDF-1免疫组化积分光密度值与供者年龄、椎间盘Thompson分级作相关分析.结果:正常组的上述5个部位SDF-1免疫组化积分光密度值分别为420.87±89.93、407.80±100.76、380.02±85.05、342.89±63.76、344.06±88.97;Western-blot灰度比值分别为0.08±0.03、0.08±0.05、0.07±0.04、0.05±0.03、0.05±0.02;rt-PCR灰度比值分别为0.22±0.06、0.22±0.05、0.20±0.04、0.20±0.04、0.20±0.04.退变组上述5个部位SDF-1免疫组化积分光密度值分别为7355.13±2271.74、5959.58±2436.23、5397.49±3044.80、1605.44±825.31、361.91±104.22; Western-blot灰度比值分别为0.55±0.29、0.52±0.28、0.42±0.18、0.21±0.12、0.06±0.04;rt-PCR灰度比值分别为0.75±0.30、0.71±0.26、0.55±0.22、0.36±0.17、0.22±0.13.退变椎间盘5个部位的SDF-1表达均高于正常组(P＜0.05),且SDF-1表达量髓核中央区＞纤维环内层＞纤维环外层＞软骨终板,差异有统计学意义(P＜0.05),而髓核中央区和髓核与纤维环交界区比较无统计学差异(P＞0.05);退变椎间盘上述5个部位的SDF-1表达与椎间盘Thompson分级、年龄呈正相关性(P＜0.05).结论:SDF-1广泛表达于退变和正常的椎间盘中,其表达在退变椎间盘中上述5个相邻部位存在差异.     

Organ culture stability of the intervertebral disc: Rat versus rabbit

There is a need to develop mechanically active culture systems to better understand the role of mechanical stresses in intervertebral disc (IVD) degeneration. Motion segment cultures that preserve the native IVD structure and adjacent vertebral bodies are preferred as model systems, but rapid ex vivo tissue degeneration limits their usefulness. The stability of rat and rabbit IVDs is of particular interest, as their small size makes them otherwise suitable for motion segment culture. The goal of this study was to determine if there are substantial differences in the susceptibility of rat and rabbit IVDs to culture‐induced degeneration. Lumbar IVD motion segments were harvested from young adult male Sprague–Dawley rats and New Zealand White rabbits and cultured under standard conditions for 14 days. Biochemical assays and safranin‐O histology showed that while glycosaminoglycan (GAG) loss was minimal in rabbit IVDs, it was progressive and severe in rat IVDs. In the rat IVD, GAG loss was concomitant with the loss of notochordal cells and the migration of endplate (EP) cells into the nucleus pulposus (NP). None of these changes were evident in the rabbit IVDs. Compared to rabbit IVDs, rat IVDs also showed increased matrix metalloproteinase‐3 (MMP‐3) and sharply decreased collagen type I and II collagen expression. Together these data indicated that the rabbit IVD was dramatically more stable than the rat IVD, which showed culture‐related degenerative changes. Based on these findings we conclude that the rabbit motion segments are a superior model for mechanobiologic studies. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 838–846, 2013     

Effect of dynamic hydrostatic pressure on rabbit intervertebral disc cells.

The pathogenesis of vibration-induced disorders of intervertebral disc at the cellular level is largely unknown. The objective of this study was to establish a method to investigate the ranges of constructive and destructive hydrostatic loading frequencies and amplitudes in preventing or inducing extracellular disc matrix degradation. Using a hydraulic chamber, normal rabbit intervertebral disc cells were tested under dynamic hydrostatic loading. Monolayer cultures of disc outer annulus cells and 3-dimensional (3-D) alginate cultures of disc nucleus pulposus cells were tested. Effects of different loading amplitudes (3-D culture, 0-3 MPa; monolayer, 0-1.7 MPa) and frequencies (1-20 Hz) on disc collagen and protein metabolism were investigated by measuring 3H-proline-labeled proteins associated with the cells in the extracellular matrix and release of 3H-proline-labeled molecules into culture medium. High frequency and high amplitude hydrostatic stress stimulated collagen synthesis in cultures of outer annulus cells whereas the lower amplitude and frequency hydrostatic stress had little effect. For the same loading duration and repetition, neither treatment significantly affected the relative amount of protein released from the cell layers, indicating that protein degradation and stability were unaffected. In the 3-D nucleus culture, higher amplitude and frequency increased synthesis rate and lowered degradation. In this case, loading amplitude had a stronger influence on cell response than that of loading frequency. Considering the ranges of loading amplitude and frequency used in this study, short-term application of high loading amplitudes and frequencies was beneficial in stimulation of protein synthesis and reduction of protein degradation.     

微RNA-210抑制人退行性变髓核细胞自噬影响椎间盘退行性变

目的探讨微RNA-210(miRNA-210)对退行性变髓核细胞自噬的影响及其参与椎间盘退行性变(IDD)进程的可能机制。方法收集24例IDD患者(IDD组)和6例腰椎骨折患者(对照组)椎间盘组织进行分离,培养髓核细胞。通过实时荧光定量聚合酶链反应(FQ-PCR)检测髓核细胞中miRNA-210的表达,通过单丹磺酰尸胺(MDC)染色和蛋白质印迹法检测细胞自噬水平。通过FQ-PCR或蛋白质印迹法检测过表达或抑制miRNA-210对细胞自噬和细胞外基质代谢的影响。通过生物信息学手段寻找miRNA-210与自噬相关的靶基因及miRNA-210的结合位点,并应用双荧光素酶报告实验验证。结果与对照组相比,IDD组髓核细胞中miRNA-210表达量增加,细胞自噬水平降低。过表达miRNA-210会抑制髓核细胞自噬,同时降低Ⅱ型胶原(ColⅡ)及蛋白多糖表达量,升高基质金属蛋白酶-3(MMP-3)和MMP-13表达量。自噬抑制剂3-MA减弱miRNA-210对ColⅡ、蛋白多糖、MMP-3和MMP-13的调节作用。miRNA-210与自噬相关蛋白7(ATG7)存在结合位点,过表达miRNA-210通过沉默ATG7抑制细胞自噬,激活MMP-3和MMP-13,促进细胞外基质(ColⅡ和蛋白多糖)降解。结论在发生退行性变的椎间盘组织中miRNA-210表达量增加。过表达的miRNA-210可能通过直接作用于靶基因ATG7抑制细胞自噬,促进细胞外基质降解,推动IDD进程。