本文關鍵詞：腰椎間盤退變動物模型的建立及四環(huán)素對腰椎間盤退變中MMP-3表達影響的研究　出處：《寧夏醫(yī)學院》2006年碩士論文　論文類型：學位論文

  更多相關文章： 椎間盤退變 基質金屬蛋白酶 四環(huán)素

【摘要】： 目的:在建立.SD大鼠腰椎間盤退變動物模型的基礎上,應用組織病理學、影像學、免疫組織化學方法、免疫印跡等方法研究MMP-3在椎間盤組織中的表達,探討MMP-3在腰椎間盤退變中的作用及四環(huán)素對退變腰椎間盤組織中MMP-3表達的影響。方法:以SD大鼠為對象,64只大鼠隨機分為正常對照組(A組)、模型對照組(B組)、四環(huán)素小劑量組(C組)、四環(huán)素大劑量組(D組),每組16只。采用手術方法以L3為中心切除L1～L6棘突、關節(jié)突、棘上、棘間韌帶,同時切斷雙側豎棘肌,建立腰椎間盤退變的動物模型,C、D組在術后2周開始給藥,C組為:25mg/kg/d; D組為:50mg/kg/d,皮下注射,持續(xù)給藥2周。建立小劑量組和大劑量組,未給藥的實驗組為模型對照組。分別于術后1、2、3月隨機選取4只SD大鼠麻醉后行X線、MRI檢查,取椎間盤組織分別行組織病理、電鏡、免疫組織化學及免疫印跡等實驗的檢測。結果:通過手術方法成功建立了腰椎間盤退變動物模型,X線檢查結果為模型對照組椎間隙狹窄,椎體不穩(wěn),軟骨終板鈣化,正常對照組無特異性改變。組織病理學結果模型對照組髓核細胞減少,外周纖維環(huán)纖維化樣變。透射電鏡顯示退變的間盤組織中髓核細胞數(shù)目減少,可見較多壞死殘跡,細胞突起減少或消失,細胞內細胞器數(shù)目和糖原顆粒明顯減少,基質中膠原原纖維發(fā)生不同程度的變性、融合、扭結或鈣化,膠原纖維束間裂隙增大。MRI提示模型對照組椎間盤T2信號減低,腦脊液白線在椎間盤部位出現(xiàn)壓跡。四環(huán)素大、小劑量組T2信號亦有輕度減低。正常對照組改變不明顯。免疫組化顯示四環(huán)素大、小劑量組MMP-3表達呈弱陽性,與模型對照組有顯著性差異(P0.05),與正常組無差異性。蛋白印跡檢測結果四環(huán)素大、小劑量組MMP-3表達水平較模型對照組低,有顯著性差異(P0.05)。結論:MMP-3在腰椎間盤退變組織中表達增高,表明MMP-3與腰椎間盤退變關系密切,并且與退變程度有相關性。四環(huán)素作為MMPs的抑制劑能降低基質金屬蛋白酶的表達,從而減輕椎間盤基質的降解,抑制和延緩腰椎間盤退變的形成和發(fā)展。為腰椎間盤退變的發(fā)生機制、預防及治療提供理論依據(jù)。
[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.
【學位授予單位】：寧夏醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R681.53;R-332

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