腰椎间盘退变动物模型的建立及四环素对腰椎间盘退变中MMP-3表达影响的研究

发布时间：2018-01-01 13:29

本文关键词：腰椎间盘退变动物模型的建立及四环素对腰椎间盘退变中MMP-3表达影响的研究　出处：《宁夏医学院》2006年硕士论文　论文类型：学位论文

【摘要】： 目的:在建立.SD大鼠腰椎间盘退变动物模型的基础上,应用组织病理学、影像学、免疫组织化学方法、免疫印迹等方法研究MMP-3在椎间盘组织中的表达,探讨MMP-3在腰椎间盘退变中的作用及四环素对退变腰椎间盘组织中MMP-3表达的影响。方法:以SD大鼠为对象,64只大鼠随机分为正常对照组(A组)、模型对照组(B组)、四环素小剂量组(C组)、四环素大剂量组(D组),每组16只。采用手术方法以L3为中心切除L1～L6棘突、关节突、棘上、棘间韧带,同时切断双侧竖棘肌,建立腰椎间盘退变的动物模型,C、D组在术后2周开始给药,C组为:25mg/kg/d; D组为:50mg/kg/d,皮下注射,持续给药2周。建立小剂量组和大剂量组,未给药的实验组为模型对照组。分别于术后1、2、3月随机选取4只SD大鼠麻醉后行X线、MRI检查,取椎间盘组织分别行组织病理、电镜、免疫组织化学及免疫印迹等实验的检测。结果:通过手术方法成功建立了腰椎间盘退变动物模型,X线检查结果为模型对照组椎间隙狭窄,椎体不稳,软骨终板钙化,正常对照组无特异性改变。组织病理学结果模型对照组髓核细胞减少,外周纤维环纤维化样变。透射电镜显示退变的间盘组织中髓核细胞数目减少,可见较多坏死残迹,细胞突起减少或消失,细胞内细胞器数目和糖原颗粒明显减少,基质中胶原原纤维发生不同程度的变性、融合、扭结或钙化,胶原纤维束间裂隙增大。MRI提示模型对照组椎间盘T2信号减低,脑脊液白线在椎间盘部位出现压迹。四环素大、小剂量组T2信号亦有轻度减低。正常对照组改变不明显。免疫组化显示四环素大、小剂量组MMP-3表达呈弱阳性,与模型对照组有显著性差异(P0.05),与正常组无差异性。蛋白印迹检测结果四环素大、小剂量组MMP-3表达水平较模型对照组低,有显著性差异(P0.05)。结论:MMP-3在腰椎间盘退变组织中表达增高,表明MMP-3与腰椎间盘退变关系密切,并且与退变程度有相关性。四环素作为MMPs的抑制剂能降低基质金属蛋白酶的表达,从而减轻椎间盘基质的降解,抑制和延缓腰椎间盘退变的形成和发展。为腰椎间盘退变的发生机制、预防及治疗提供理论依据。
[Abstract]:Objective: to establish an animal model of lumbar disc degeneration in SD rats and to apply histopathological, imaging and immunohistochemical methods. The expression of MMP-3 in intervertebral disc tissue was studied by Western blot. To investigate the role of MMP-3 in lumbar disc degeneration and the effect of tetracycline on the expression of MMP-3 in degenerative lumbar intervertebral disc. 64 rats were randomly divided into normal control group (group A), model control group (group B), tetracycline group (group C) and tetracycline group (group D). 16 rats in each group were treated with L3 as the center of resection of spinous process, articular process, supraspinal ligament and interspinous ligament, and bilateral erector spinous muscles were cut off at the same time, and the animal model of lumbar disc degeneration was established. 2 weeks after operation, group D was given drug at 1: 25 mg / kg / d in group C; Group D: 50 mg / kg / d, subcutaneously injected for 2 weeks. Small dose group and large dose group were established. The experimental group without administration was the model control group. In March, 4 Sprague-Dawley (SD) rats were randomly selected for X-ray and MRI examination after anesthesia. Histopathological and electron microscopy were performed in the intervertebral disc tissue. Results: the animal model of lumbar intervertebral disc degeneration was successfully established by operation. The results of X-ray examination were as follows: the vertebral space was narrow and the vertebral body was unstable in the model control group. Cartilage endplate calcification, normal control group had no specific changes. Histopathological results showed that the number of nucleus pulposus cells decreased in the model control group. Transmission electron microscopy showed that the number of nucleus pulposus cells in the degenerated disc tissue was decreased, and more necrotic traces were observed, and the cell processes decreased or disappeared. The number of cell organelles and glycogen granules decreased significantly, and collagen fibers in the matrix were denatured, fused, kink or calcified to varying degrees. The enlargement of interfascicular fissure of collagen fibers. MRI indicated that the T2 signal of intervertebral disc in the model control group was decreased, the white line of cerebrospinal fluid appeared pressure trace in the intervertebral disc, and the tetracycline was large. The T 2 signal in the low dose group was also slightly decreased. The normal control group had no obvious change. Immunohistochemistry showed that tetracycline was large and the MMP-3 expression in the low dose group was weakly positive. There was significant difference between the model control group and the model control group (P 0.05), but there was no difference between the model group and the normal group. Western blot analysis showed that tetracycline was large, and the MMP-3 expression level in the low dose group was lower than that in the model control group. There was significant difference (P 0.05). Conclusion the expression of MMP-3 in lumbar intervertebral disc degeneration tissue is increased, indicating that MMP-3 is closely related to lumbar disc degeneration. Tetracycline, as an inhibitor of MMPs, can reduce the expression of matrix metalloproteinases and reduce the matrix degradation of intervertebral disc. To inhibit and delay the formation and development of lumbar disc degeneration, provide theoretical basis for the mechanism, prevention and treatment of lumbar disc degeneration.
【学位授予单位】：宁夏医学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R681.53;R-332

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