IL-1α及BMP-7在黄韧带肥厚引起的腰椎管狭窄症中的表达研究

发布时间：2018-04-28 02:11

本文选题：IL-1α + BMP-7　；参考：《川北医学院》2015年硕士论文

【摘要】：目的:通过检测IL-1α及BMP-7在人类腰椎退变性黄韧带肥厚中的表达情况,探索其在黄韧带肥厚中的作用机制,为黄韧带肥厚引起的腰椎狭窄症及黄韧带骨化的干预提供新的思路。方法:1.收集2013年1月—2014年10月在川北医学院附属医院骨科住院治疗的腰椎骨折和腰椎管狭窄患者黄韧带标本共20例。其中腰椎骨折患者10例为对照组,年龄18-30岁,男8例,女2例,既往健康,排除其他慢性疾病,经影像学证实无腰椎黄韧带肥厚;腰椎管狭窄症患者10例为实验组,年龄55-65岁,男4例,女6例,既往健康,排除其他慢性疾病,经影像学证实腰椎黄韧带0.5cm。2.术中取椎板间整块黄韧带,测量标本厚度,常规石蜡切片,HE染色观察成纤维细胞并计数,免疫组织化学法、RT-PCR及Western blotting检测实验组和对照组中黄韧带IL-1α及BMP-7,各组测得的数据均用均数±标准差(sx±)表示,用SPSS19.0软件分析包进行统计学分析,计数资料卡方检验,计量资料t检验P0.05认为差异具有统计学意义。结果:1.大体标本形态学观察:对照组标本表面光滑、透亮、柔软,和周边组织边界清楚,易于分离,切下后回缩快,呈卷曲状,弹性良好,均数标准差0.215±0.37(cm)。实验组标本表面欠光滑、周围粘连明显,质地致密、坚韧、不易分离,未发现钙化和骨化,均数标准差0.678±0.064(cm)。两组间比较差异有统计学意义P0.05。2.HE染色:对照组黄韧带纤维大小相近,长轴排列,表面光滑,纤维较细,呈波浪状,其主要成分为弹力纤维,纤维方向与黄韧带的长轴平行,并有分支发出,相互连接,形成网状结构;成纤维细胞计数27.00±5.18/HP。实验组黄韧带纤维粗细不均,排列紊乱,纤维断裂,弹性纤维减少,结构变形,功能活跃的成纤维细胞增多,未发现钙化及骨化;成纤维细胞计数74.40±7.26/HP。两组间比较差异有统计学意义(P0.05)。3.免疫组织化学法检测:IL-1α在对照组和实验组阳性细胞百分比分别为40%和90%,BMP-7在对照组和实验组阳性细胞百分比分别为10%和90%,两组比较差异有统计学意义(P0.05)。4.实时荧光定量PCR法检测:各黄韧带标本中基因扩增曲线、融解曲线良好。黄韧带组织学研究发现,IL-1α和BMP-7m RNA在实验组中的表达均明显高于对照组(P0.05)。5.Western blot检测:实验组IL-1α和BMP-7的表达水平均较对照组显著增强,差异有统计学意义(P0.05)。结论:1.IL-1α表达增加可能是导致黄韧带退变的重要机制之一2.BMP-7表达增加可能是导致黄韧带退变的重要机制之一。
[Abstract]:Objective: to investigate the expression of IL-1 伪 and BMP-7 in the hypertrophy of degenerative ligamentum flavum in human lumbar vertebrae, and to explore the mechanism of its role in the hypertrophy of ligamentum flavum, so as to provide a new idea for the intervention of lumbar stenosis and ossification of ligamentum flavum caused by hypertrophy of ligamentum flavum. Method 1: 1. From January 2013 to October 2014, 20 specimens of ligamentum flavum were collected from lumbar fractures and lumbar spinal stenosis treated in orthopaedic department of affiliated Hospital of North Sichuan Medical College. There were 10 cases of lumbar vertebrae fracture as control group, aged 18-30 years, male 8 cases, female 2 cases, past health, excluding other chronic diseases, no lumbar ligamentum flavum hypertrophy was confirmed by imaging, and 10 cases of lumbar spinal stenosis were treated as experimental group, aged 55-65 years. Male 4 cases, female 6 cases, past health, excluding other chronic diseases, lumbar ligamentum flavum 0.5 cm. 2. The interlaminar ligamentum flavum was taken during the operation and the thickness of the specimen was measured. The fibroblasts were observed and counted by HE staining in routine paraffin sections. Immunohistochemical method was used to detect IL-1 伪 and BMP-7 in experimental and control groups by RT-PCR and Western blotting. The data of each group were expressed as mean 卤standard deviation (sx 卤). Statistical analysis was carried out with SPSS19.0 software analysis kit, and counting data was chi-square test. Measurement data t test P0.05 that the difference was statistically significant. The result is 1: 1. Morphological observation of gross specimens: the surface of the control group was smooth, transparent and soft, and the border with the surrounding tissue was clear and easy to be separated. After cutting, the specimen was curled, with good elasticity, and the mean standard deviation was 0.215 卤0.37 cm ~ (-1) 路cm ~ (-1). In the experimental group, the surface of the specimen was not smooth, the adhesion around it was obvious, the texture was dense, tough and difficult to separate, no calcification and ossification were found, and the mean standard deviation was 0.678 卤0.064 cm ~ (-1). There were significant differences in P0.05.2.HE staining between the two groups: in the control group, the fibers of the ligamentum flavum were similar in size, arranged in long axis, smooth in surface, fine in fiber and wavy in shape. The main component of the fibers was elastic fibers, and the direction of the fibers was parallel to the long axis of the ligamentum flavum. The number of fibroblasts was 27.00 卤5.18% HP. In the experimental group, the fibers of ligamentum flavum were uneven in thickness, disordered in arrangement, broken in fiber, decreased in elastic fiber, deformed in structure, increased in number of active fibroblasts, no calcification and ossification were found, and the number of fibroblasts was 74.40 卤7.26% HP. The difference between the two groups was statistically significant (P 0.05). The percentage of positive cells in control group and experimental group were 40% and 90% respectively by immunohistochemical method. The percentage of positive cells of BMP-7 in control group and experimental group were 10% and 90%, respectively. The difference between the two groups was statistically significant. Real-time fluorescence quantitative PCR assay showed that the curve of gene amplification in ligamentum flavum was good. Histological study of ligamentum flavum showed that the expressions of IL-1 伪 and BMP-7m RNA in the experimental group were significantly higher than those in the control group (P 0.05A). 5. Western blot: the expression levels of IL-1 伪 and BMP-7 in the experimental group were significantly higher than those in the control group (P 0.05). Conclusion 1. The increased expression of IL-1 伪 may be one of the important mechanisms leading to the degeneration of ligamentum flavum. The increase of 2.BMP-7 expression may be one of the important mechanisms leading to degeneration of ligamentum flavum.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R681.5

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