椎间盘炎动物模型的建立及自免疫状态的研究

发布时间：2018-05-15 06:46

本文选题：椎间盘炎 + 自免疫反应　；参考：《天津医科大学》2007年硕士论文

【摘要】： 目的：通过手术破坏纤维环和软骨终板建立椎间盘炎的动物模型，探讨其影像学表现及自免疫状态表达的变化过程，探讨自免疫反应在椎间盘炎中的作用机制，为临床诊断和治疗提供理论基础。方法：选择健康成年新西兰大耳白兔50只，雌雄不限，随机分为三组：手术组(30只)，经前外侧入路手术破坏腰椎间盘纤维环及软骨终板，建立兔椎间盘炎动物模型；设立假手术组(10只)及对照组(10只)。对照组即刻行X线及MRI检查，处死取腰3／4、腰4／5、腰5／6椎间盘标本。假手术组同手术组取相同手术入路，暴露椎间盘但不破坏纤维环和软骨终板。手术组与假手术组分别于术后1周、2周、4周、8周、12周分批处死动物，处死取材前行X线片、MRI检查。取手术区椎间盘标本，行：(1)常规HE染色，形态学比较及淋巴细胞的表达；(2)Masson染色，观察肌组织炎症纤维化表现；(3)应用免疫组化技术和免疫荧光技术观察标本CD4阳性、CD8阳性T淋巴细胞，免疫球蛋白IgG、IgM的表达。手术组与假手术组、对照组进行比较以分析各指标的表达及变化过程，探讨自免疫反应在椎间盘炎中的作用：(4)细菌学培养。结果：1．影像学表现：手术组常规X线检查在术后1-2周未见异常征象，术后4周显示手术区椎间隙狭窄，术后12周可见椎体骨质破坏和硬化。术后4周MRI矢状位T2WI上即可见手术椎间盘信号异常，呈不同程度的破坏、碎裂、变小，正常髓核的高信号消失，呈低信号改变。终板及终板下椎体松质骨有不同程度的侵犯破坏致椎体和椎间盘分界模糊不清，相邻椎体受累，T2WI上呈混杂斑点状高信号。术后12周T2WI上可见手术椎间盘相邻椎体高信号，发生病理性骨折。假手术组与对照组均未见异常。2．HE染色：光镜下可见手术组椎间盘内、外层纤维环紊乱，失去正常结构，胶原纤维不连续、增生，外层纤维环成纤维细胞增多，术后1-4周可见淋巴细胞浸润，近纤维环内层髓核部分有大量软骨细胞增生。假手术组及对照组则无上述表现。3．Masson染色：光镜下可见手术组椎间盘外层纤维环呈绿色，其边缘肌组织呈红色，两者交界的灰色区域为肌组织因炎症反应而正纤维化。其间散在淋巴细胞。4．免疫组化染色：手术组1周、2周、4周的标本可见近边缘细胞浸润区有CD4阳性、CD8阳性淋巴细胞分布，其中CD4阳性淋巴细胞主要在术后1周、2周取材的标本表达较多，CD8阳性淋巴细胞则在术后2周、4周取材的标本表达较多。而假手术组则于术后1周、2周可见有极少量淋巴细胞表达，对照组未见淋巴细胞表达。手术组淋巴细胞表达与假手术组、对照组比较具有统计学意义。5．免疫荧光：手术组可见手术椎间盘近边缘区免疫球蛋白IgG、IgM呈阳性反应，于1周少量表达，术后2周、4周、8周大量表达。假手术组及对照组均可见IgG极少量表达，未见IgM表达。手术组与假手术组、对照组比较具有统计学意义。6．细菌培养阴性。结论：1．运用前外侧入路手术破坏纤维环及软骨终板可以建立椎间盘炎动物模型，其影像学表现符合临床文献对椎间盘炎的报道；2．纤维环及软骨终板受破坏后，，髓核等自身抗原可与血液循环接触引发机体自身免疫反应，可表现在细胞免疫和体液免疫两方面；3．自免疫反应在椎间盘炎的发病机制和病理演变过程以及症状产生过程中起重要作用。
[Abstract]:Objective: to establish an animal model of intervertebral disc inflammation by destroying the fibrous ring and cartilage endplate, to explore the changes in the expression of the image and the expression of self immune state, and to explore the mechanism of the action of self immunization in the disease of the intervertebral disc, and provide a theoretical basis for the clinical diagnosis and treatment.
Methods: 50 healthy adult New Zealand white rabbits were randomly divided into three groups: the operation group (30), the anterior lateral approach was used to destroy the lumbar intervertebral disc and cartilage endplate, and the animal model of rabbit's disc inflammation was established, and the sham operation group (10) and the contrast group (10 rats) were set up. The control group was executed immediately by X-ray and MRI examination. Lumbar 3 / 4, waist 4 / 5, lumbar 5 / 6 disc specimens. The sham operation group took the same surgical approach with the operation group, exposed the intervertebral disc but did not destroy the fibrous ring and cartilage endplate. The operation group and the sham operation group were executed 1 weeks, 2 weeks, 4 weeks, 8 weeks, and 12 weeks respectively. Before the operation, the X-ray film and MRI examination were performed. The surgical area disc specimens were taken: (1) often: (1) often HE staining, morphological comparison and expression of lymphocyte, (2) Masson staining, observation of inflammatory and fibrosis of muscle tissue; (3) immunohistochemical technique and immunofluorescence technique were used to observe the CD4 positive, CD8 positive T lymphocyte, immunoglobulin IgG, IgM expression. The comparison of the indexes between the hand group and the sham operation group and the control group was compared to analyze the indexes. To explore the role of autoimmune response in intervertebral disc inflammation: (4) bacteriological culture.
Results: 1. imaging findings: no abnormal signs were found at 1-2 weeks after operation in the operation group. The intervertebral space in the surgical area was narrowed at 4 weeks after the operation. The vertebral bone destruction and hardening were observed at 12 weeks after the operation. The abnormal signal of the surgical intervertebral disc was seen on the MRI sagittal T2WI at 4 weeks after the operation. The signal disappeared and showed a low signal change. The vertebral and intervertebral discs in the end plate and the inferior end of the end plate had varying degrees of invasion and destruction of the vertebral body and intervertebral disc, and the adjacent vertebrae were involved, and the T2WI showed a high signal of mixed spot. 12 weeks after the operation, the high signal of the adjacent vertebral body was observed on the surgical intervertebral disc, and the pathological fracture occurred. The sham operation group and the control group were both. No abnormal.2.HE staining was found in the intervertebral disc in the surgical group. The outer fibrous ring was disordered, the normal structure was lost, the collagen fibers were discontinuous, the proliferation of the fibrous cell, the proliferation of the outer fibroblast cells, the infiltration of the lymphocytes in the 1-4 weeks after the operation, and the proliferation of a large number of chondrocytes in the nucleus of the inner fibrous ring. The sham operation group and the control group were not. .3.Masson staining showed that the fibrous ring in the outer layer of the intervertebral disc in the operation group was green, its marginal muscle tissue was red, and the gray area of the junction was positive fibrosis of the muscle tissue due to the inflammatory reaction. In the meantime, it scattered in the lymphocyte.4. immunohistochemical staining: the surgical group was 1 weeks, 2 weeks, and 4 weeks showed the infiltration of the near marginal cells. There were CD4 positive and CD8 positive lymphocytes in the area, in which CD4 positive lymphocytes were mainly expressed at 1 weeks after operation and more expressed in 2 weeks, while CD8 positive lymphocytes were more expressed in 2 weeks after the operation and 4 weeks after the operation, while in the sham operation group, a very small amount of lymphocyte expression was found at 1 weeks and 2 weeks after the operation, and no lymphocyte expression was found in the control group. The expression of lymphocyte in the operation group and the sham operation group and the control group were statistically significant.5. immunofluorescence: the operation group showed the immunoglobulin IgG in the near marginal area of the surgical intervertebral disc, the positive reaction of IgM, and a small amount of expression at 1 weeks, 2 weeks, 4 weeks, and 8 weeks after the operation. No IgM table was found in the sham operation group and the control group. No IgM table was found in the sham operation group and the control group. The operation group and the sham operation group had statistical significance compared with the control group..6. bacteria culture was negative.
Conclusion: 1. the animal model of intervertebral disc inflammation can be established by using the anterolateral approach to destroy the fibrous rings and cartilage endplates. The imaging findings are in line with the report of the clinical literature on the intervertebral disc inflammation. 2. after the destruction of the fibrous rings and cartilage endplates, the autoantigens such as the nucleus pulposus can trigger the body's autoimmune reaction with the blood circulation, which may be shown in the body. Two aspects of cell immunity and humoral immunity; 3. the autoimmune reaction plays an important role in the pathogenesis, pathological process and symptom process of intervertebral disc inflammation.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R681.53

【参考文献】