HMGB1介导MAPKs信号通路影响椎间盘软骨终板退变过程及其机制研究

Lumbocrural pain is the most common diseases in the department of orthopaedics and pain. These are the main reasons that induce decline of activity and loss of labor force in daily life. These are common in the elders, but the scale in youngth grows higher recently. To present, intervertebral disc degeneration has been considered as the most common reason in Lumbocrural pain.The nutrition of intervertebral disc was contributed by the cartilage endpalte. As a result, the important role of the cartilage endplate in intervertebral disc degeneration has been valued. In rescent researches, the process of cartilage endplate degeneration involved the homeostasis of various proteins and factors, but the underlying interlink and mechanism was still unclear. This project will establish an animal intervetebral destabilization model which may induce cartilage endplate degeneration.Based on this model, imageological, histochemical, cellular biological and molecular biologial method will be performed to exam HMGB1 regulates expression of TNF-a, IL-1β, MMP3 and VEGF by MAPKs pathway, For illustrateing their underlying regulatory mechanism. By blocking HMGB1 and MAPKs pathway, observation to prevent and delay the effect of cartilage endplate degeneration, thus apply novel therapeutic strategy in blockade or postpone of intervetebral disc degeration.

腰腿痛是骨科和疼痛科室常见疾病，是许多人活动能力下降和劳动力丧失的起因。在中老年人群中常见，近年有年轻化的趋势。目前很多学者认为椎间盘退变是腰腿痛最常见的原因，椎间盘的营养途径主要是软骨终板途径，因此软骨终板在椎间盘退变中的作用日益被重视。最近的研究发现，软骨终板退变过程中涉及多种蛋白质及细胞因子的变化，但它们之间的相互作用机理尚不明确。本项目通过复制椎间失稳导致软骨终板退变的动物模型，利用影像学、组织化学、细胞生物学、分子生物学技术等多种观察和检测手段，研究软骨终板退变过程中HMGB1通过MAPKs通路调节TNF-a、IL-1β、MMP3、VEGF等因子的表达规律，以阐明HMGB1在软骨终板退变中的作用及机理。通过阻断HMGB1及MAPKs通路，观察阻止和延缓软骨终板退变的效果，为延缓或治疗椎间盘退变提供新的思路。

软骨终板的炎性反应是造成该结构退变的主要原因，而软骨终板的退变在腰椎间盘退行性疾病的发病机制中起着关键性的作用。虽然HMGB1（high mobility group box-1 protein高迁移率蛋白-1）与软骨终板细胞的退变密切相关，但是具体的机制仍不清楚。我们通过研究SD大鼠腰椎软骨终板细胞内HMGB1与基质金属肽酶-3（MMP-3），白细胞介素-10（IL-10）和血管内皮生长因子（VEGF）的关系，来探讨HMGB1在大鼠腰椎软骨终板退变中所起的作用。.实验中分别使用RT-PCR、Western blot和Elisa从mRNA和蛋白质水平检测MMP-3、VEGF和IL-10的变化以及评估ERK和JNK信号转导途径磷酸化的变化。在大鼠腰椎软骨终板细胞中，我们的实验结果得出HMGB1可以通过激活ERK和 JNK信号转导途径增加MMP-3和VEGF的表达以及通过激活JNK信号转导途径降低IL-10的表达从而导致软骨终板的退变。 .在另外的实验中表明，兔腰椎间盘软骨终板中HMGB1与VEGF的持续升高与椎间盘退变加重有关。.我们的实验结果阐明了HMGB1介导MAPKs引起软骨终板退变的分子机制。而国内外的文献对此的报道还比较少。本研究为临床治疗中采用抑制HMGB1作用软骨终板炎性途径和机理的方法来阻止或预防腰椎椎间盘退变提供理论依据，也为临床预防或阻止椎间盘退变性疾病提供了一个新的思路。