基质金属蛋白酶-9对多发性硬化复发诊断的临床意义研究

发布时间：2018-06-18 06:37

本文选题：基质金属蛋白酶 + 9　；参考：《苏州大学》2015年硕士论文

【摘要】：1、目的:多发性硬化(Multiple sclerosis.MS)是一种中枢神经系统慢性炎症脱髓鞘疾病,它的典型病理特征是炎性细胞浸润、髓鞘脱失、轴索变性以及胶质细胞增生等,目前许多研究表明多发性硬化早期即存在血脑屏障破坏。基质金属蛋白酶-9(Matrix metalloproteinases-9.MMP-9)是一类锌离子依赖的蛋白酶,具有破坏血脑屏障,降解细胞外基质、髓鞘碱性蛋白的作用,可以导致血管内皮细胞通透性增高,进而导致T淋巴细胞等炎症细胞穿透血管基底膜进入中枢神经系统。MS血脑屏障的破坏、炎性细胞浸润、髓鞘降解以及轴索的损伤等主要病理过程均与MMP-9活性异常增高有关。颈椎病神经根型发病率为50-60%,是由于颈椎间盘向侧后方突出、关节增生及肥大,刺激或压迫神经根所致。临床上开始多为颈肩痛,并向上肢放射,皮肤可有麻木、过敏等感觉异常,颈椎病神经根型临床表现与多发性硬化颇为相似。血脑屏障破坏在MS的病理生理中起重要作用,血管通透性的改变早于MS影像学的特征性变化。核磁共振(MRI)增强扫描可见活动性病灶强化,这与病变血管周围炎性改变,血脑屏障破坏,血管通透性升高,造影剂外渗有关,MMP-9是引起血脑屏障破坏的重要因素,MS患者外周血MMP-9突然升高与新病灶的形成有关,故推断MS复发期较缓解期的MMP-9表达不同。本研究旨在探讨基质金属蛋白酶-9(MMP-9)的检测能否成为自身免疫性疾病多发性硬化(MS)复发-缓解型(R-R型)和颈椎病神经根型相鉴别的一种方法,以及MMP-9的检测对判断MS的复发和缓解是否具有临床意义。2、方法:2.1研究分组:本研究分别对经CT检查、肌电图检查及核磁共振扫描已确诊的MS(R-R型)患者26例(复发期和缓解期),颈椎病神经根型患者30例(复发期和缓解期),正常对照组30例共5组分别抽取不加抗凝的静脉血2ml送MMP-9检测。2.2试验仪器:MMP-9检测所选用的设备主要有酶标仪、洗板机、数字显示隔水式电热恒温培养箱、离心机、移液枪、漩涡混合器、烘箱、分液器及微量电动组织匀浆器等。2.3试验过程:本实验采用双抗体夹心ABC-ELISA法。用抗人MMP-9单抗包被于酶标板上,标准品和样品中的MMP-9与单抗结合,加入生物素化的抗人MMP-9,形成免疫复合物连接在板上,辣根过氧化物酶标记的Streptavidin与生物素结合,加入底物工作液显蓝色,最后加终止液硫酸,在450nm处测OD值,MMP-9浓度与OD值成正比,可通过绘制标准曲线求出标本中MMP-9浓度,将所获得的数据再进行统计学分析。3、结果:3.1 MS(R-R型)患者其复发期和缓解期的MMP-9分别与正常对照组MMP-9比较均有统计学差异。3.2 颈椎病神经根型患者其复发期和缓解期的MMP-9分别与正常对照组MMP-9比较没有统计学差异。3.3 MS(R-R型)患者其复发期和缓解期的MMP-9分别与颈椎病神经根型患者其复发期和缓解期的MMP-9比较均有统计学差异。3.4 MS患者的复发期与缓解期的MMP-9比较,有统计学差异。4、结论4.1 MMP-9的检测对于MS(R-R型)和颈椎病神经根型的鉴别具有临床意义。4.2对于MS(R-R型)患者定期复查MMP-9可以早期判断及预防有无疾病复发,为积极诊断及治疗提供帮助。4.3 MMP-9的检测对于颈椎病神经根型患者无现实临床意义。
[Abstract]:1, objective: multiple sclerosis (Multiple sclerosis.MS) is a chronic inflammatory demyelinating disease of the central nervous system. Its typical pathological features are inflammatory cell infiltration, myelin loss, axon degeneration, and glial cell proliferation. Many studies have shown that there is blood brain barrier damage at the early stage of multiple harden. Matrix metalloproteinase -9 (M) Atrix metalloproteinases-9.MMP-9) is a kind of zinc ion dependent protease, which destroys the blood brain barrier, degrades the extracellular matrix and myelin basic protein, which can lead to the increase of the permeability of the vascular endothelial cells, and then the T lymphocyte and other inflammatory cells penetrate the vascular basement membrane into the.MS blood brain barrier of the central nervous system. The main pathological processes, such as bad, inflammatory cell infiltration, myelin degradation and axonal injury, are related to the abnormal increase of MMP-9 activity. The incidence of nerve root type of cervical spondylosis is 50-60%, which is caused by the protruding of the cervical intervertebral disc to the side of the side, the proliferation and hypertrophy of the joints, and the stimulation or compression of the nerve roots. The skin can have numbness, allergies and other sensory abnormalities. The clinical manifestation of the cervical spondylosis is quite similar to that of multiple sclerosis. Blood brain barrier damage plays an important role in the pathophysiology of MS. The changes of vascular permeability are earlier than the characteristic changes in MS imaging. Nuclear magnetic resonance (MRI) enhanced scan shows the enhancement of active lesion, which is associated with the perivascular perivascular disease. Perin-inflammatory changes, blood brain barrier destruction, vascular permeability and contrast exootion are related, MMP-9 is an important factor causing the destruction of blood brain barrier. The sudden increase of MMP-9 in peripheral blood of patients with MS is related to the formation of new lesions. Therefore, it is concluded that the expression of MMP-9 in the recurrence period of MS is different than that in the remission stage. This study aims to explore the detection of matrix metalloproteinase (MMP-9). Whether or not it can be a method to identify the relapsing remission (type R-R) and cervical spondylosis of the autoimmune disease (MS) and the cervical spondylosis, and whether the detection of MMP-9 is of clinical significance to determine the recurrence and remission of MS. The 2.1 study groups: This study was divided into CT examination, electromyography examination and MRI scans. 26 patients with MS (R-R type) (relapse and remission), 30 cases of cervical spondylosis and nerve root type patients (relapse and remission period), 30 cases of normal control group, 30 cases of non anticoagulant venous blood 2ml MMP-9 test instrument for.2.2 test: the equipment selected for MMP-9 detection should have an enzyme labeling apparatus, a washing machine, and a digital display of water insulating Heng Wenpei. .2.3 test process of box, centrifuge, centrifuge, liquid gun, whirlpool mixer, oven, liquid separator and micro electro tissue homogenizer, and so on. This experiment uses the double antibody sandwich ABC-ELISA method. Using the anti human MMP-9 monoclonal antibody package on the enzyme standard plate, the standard product and the sample MMP-9 and the monoclonal antibody, join the biotinylated anti human MMP-9 and form the immune complex connection. On the board, the Streptavidin of horseradish peroxidase combined with biotin, adding the substrate working liquid to show blue, and finally adding terminating liquid sulphuric acid, measuring the OD value at 450nm, the MMP-9 concentration is proportional to the OD value, and the MMP-9 concentration in the specimen can be calculated by drawing the standard curve, and the obtained data are then statistically analyzed.3, the result: 3.1 MS (R-R type). The recurrence period and remission phase of the patients with MMP-9 were respectively compared with the normal control group MMP-9. The recurrence period and the remission period of MMP-9 in the patients with cervical spondylosis were not statistically different from that of the normal control group MMP-9, respectively,.3.3 MS (R-R type), and the recurrence and remission phase of MMP-9 in the patients with cervical spondylosis and the cervical spondylosis nerve root type respectively. There were statistically significant differences in the recurrence period and remission period of MMP-9 in patients with.3.4 MS, compared with MMP-9 in remission period, there was a statistically significant difference in.4. Conclusion 4.1 MMP-9 has a clinical significance for the identification of MS (R-R type) and cervical spondylosis nerve root type,.4.2 for MS (R-R type) patients, regular review of MMP-9 can be used for early judgment and prevention. There is no relapse of disease, which is helpful for active diagnosis and treatment. The detection of.4.3 MMP-9 has no real clinical significance for patients with cervical spondylotic radicular type.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R744.51

【相似文献】