發(fā)布時間：2018-05-15 06:46

  本文選題：椎間盤炎 + 自免疫反應(yīng)��； 參考：《天津醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 目的：通過手術(shù)破壞纖維環(huán)和軟骨終板建立椎間盤炎的動物模型，探討其影像學(xué)表現(xiàn)及自免疫狀態(tài)表達的變化過程，探討自免疫反應(yīng)在椎間盤炎中的作用機制，為臨床診斷和治療提供理論基礎(chǔ)。 方法：選擇健康成年新西蘭大耳白兔50只，雌雄不限，隨機分為三組：手術(shù)組(30只)，經(jīng)前外側(cè)入路手術(shù)破壞腰椎間盤纖維環(huán)及軟骨終板，建立兔椎間盤炎動物模型；設(shè)立假手術(shù)組(10只)及對照組(10只)。對照組即刻行X線及MRI檢查，處死取腰3／4、腰4／5、腰5／6椎間盤標(biāo)本。假手術(shù)組同手術(shù)組取相同手術(shù)入路，暴露椎間盤但不破壞纖維環(huán)和軟骨終板。手術(shù)組與假手術(shù)組分別于術(shù)后1周、2周、4周、8周、12周分批處死動物，處死取材前行X線片、MRI檢查。取手術(shù)區(qū)椎間盤標(biāo)本，行：(1)常規(guī)HE染色，形態(tài)學(xué)比較及淋巴細胞的表達；(2)Masson染色，觀察肌組織炎癥纖維化表現(xiàn)；(3)應(yīng)用免疫組化技術(shù)和免疫熒光技術(shù)觀察標(biāo)本CD4陽性、CD8陽性T淋巴細胞，免疫球蛋白IgG、IgM的表達。手術(shù)組與假手術(shù)組、對照組進行比較以分析各指標(biāo)的表達及變化過程，探討自免疫反應(yīng)在椎間盤炎中的作用：(4)細菌學(xué)培養(yǎng)。 結(jié)果：1．影像學(xué)表現(xiàn)：手術(shù)組常規(guī)X線檢查在術(shù)后1-2周未見異常征象，術(shù)后4周顯示手術(shù)區(qū)椎間隙狹窄，術(shù)后12周可見椎體骨質(zhì)破壞和硬化。術(shù)后4周MRI矢狀位T2WI上即可見手術(shù)椎間盤信號異常，呈不同程度的破壞、碎裂、變小，正常髓核的高信號消失，呈低信號改變。終板及終板下椎體松質(zhì)骨有不同程度的侵犯破壞致椎體和椎間盤分界模糊不清，相鄰椎體受累，T2WI上呈混雜斑點狀高信號。術(shù)后12周T2WI上可見手術(shù)椎間盤相鄰椎體高信號，發(fā)生病理性骨折。假手術(shù)組與對照組均未見異常。2．HE染色：光鏡下可見手術(shù)組椎間盤內(nèi)、外層纖維環(huán)紊亂，失去正常結(jié)構(gòu)，膠原纖維不連續(xù)、增生，外層纖維環(huán)成纖維細胞增多，術(shù)后1-4周可見淋巴細胞浸潤，近纖維環(huán)內(nèi)層髓核部分有大量軟骨細胞增生。假手術(shù)組及對照組則無上述表現(xiàn)。3．Masson染色：光鏡下可見手術(shù)組椎間盤外層纖維環(huán)呈綠色，其邊緣肌組織呈紅色，兩者交界的灰色區(qū)域為肌組織因炎癥反應(yīng)而正纖維化。其間散在淋巴細胞。4．免疫組化染色：手術(shù)組1周、2周、4周的標(biāo)本可見近邊緣細胞浸潤區(qū)有CD4陽性、CD8陽性淋巴細胞分布，其中CD4陽性淋巴細胞主要在術(shù)后1周、2周取材的標(biāo)本表達較多，CD8陽性淋巴細胞則在術(shù)后2周、4周取材的標(biāo)本表達較多。而假手術(shù)組則于術(shù)后1周、2周可見有極少量淋巴細胞表達，對照組未見淋巴細胞表達。手術(shù)組淋巴細胞表達與假手術(shù)組、對照組比較具有統(tǒng)計學(xué)意義。5．免疫熒光：手術(shù)組可見手術(shù)椎間盤近邊緣區(qū)免疫球蛋白IgG、IgM呈陽性反應(yīng)，于1周少量表達，術(shù)后2周、4周、8周大量表達。假手術(shù)組及對照組均可見IgG極少量表達，未見IgM表達。手術(shù)組與假手術(shù)組、對照組比較具有統(tǒng)計學(xué)意義。6．細菌培養(yǎng)陰性。 結(jié)論：1．運用前外側(cè)入路手術(shù)破壞纖維環(huán)及軟骨終板可以建立椎間盤炎動物模型，其影像學(xué)表現(xiàn)符合臨床文獻對椎間盤炎的報道；2．纖維環(huán)及軟骨終板受破壞后，，髓核等自身抗原可與血液循環(huán)接觸引發(fā)機體自身免疫反應(yīng)，可表現(xiàn)在細胞免疫和體液免疫兩方面；3．自免疫反應(yīng)在椎間盤炎的發(fā)病機制和病理演變過程以及癥狀產(chǎn)生過程中起重要作用。
[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.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R-332;R681.53

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