本文選題：基質(zhì)金屬蛋白酶 + 9　； 參考：《蘇州大學(xué)》2015年碩士論文

【摘要】：1、目的:多發(fā)性硬化(Multiple sclerosis.MS)是一種中樞神經(jīng)系統(tǒng)慢性炎癥脫髓鞘疾病,它的典型病理特征是炎性細胞浸潤、髓鞘脫失、軸索變性以及膠質(zhì)細胞增生等,目前許多研究表明多發(fā)性硬化早期即存在血腦屏障破壞�；|(zhì)金屬蛋白酶-9(Matrix metalloproteinases-9.MMP-9)是一類鋅離子依賴的蛋白酶,具有破壞血腦屏障,降解細胞外基質(zhì)、髓鞘堿性蛋白的作用,可以導(dǎo)致血管內(nèi)皮細胞通透性增高,進而導(dǎo)致T淋巴細胞等炎癥細胞穿透血管基底膜進入中樞神經(jīng)系統(tǒng)。MS血腦屏障的破壞、炎性細胞浸潤、髓鞘降解以及軸索的損傷等主要病理過程均與MMP-9活性異常增高有關(guān)。頸椎病神經(jīng)根型發(fā)病率為50-60%,是由于頸椎間盤向側(cè)后方突出、關(guān)節(jié)增生及肥大,刺激或壓迫神經(jīng)根所致。臨床上開始多為頸肩痛,并向上肢放射,皮膚可有麻木、過敏等感覺異常,頸椎病神經(jīng)根型臨床表現(xiàn)與多發(fā)性硬化頗為相似。血腦屏障破壞在MS的病理生理中起重要作用,血管通透性的改變早于MS影像學(xué)的特征性變化。核磁共振(MRI)增強掃描可見活動性病灶強化,這與病變血管周圍炎性改變,血腦屏障破壞,血管通透性升高,造影劑外滲有關(guān),MMP-9是引起血腦屏障破壞的重要因素,MS患者外周血MMP-9突然升高與新病灶的形成有關(guān),故推斷MS復(fù)發(fā)期較緩解期的MMP-9表達不同。本研究旨在探討基質(zhì)金屬蛋白酶-9(MMP-9)的檢測能否成為自身免疫性疾病多發(fā)性硬化(MS)復(fù)發(fā)-緩解型(R-R型)和頸椎病神經(jīng)根型相鑒別的一種方法,以及MMP-9的檢測對判斷MS的復(fù)發(fā)和緩解是否具有臨床意義。2、方法:2.1研究分組:本研究分別對經(jīng)CT檢查、肌電圖檢查及核磁共振掃描已確診的MS(R-R型)患者26例(復(fù)發(fā)期和緩解期),頸椎病神經(jīng)根型患者30例(復(fù)發(fā)期和緩解期),正常對照組30例共5組分別抽取不加抗凝的靜脈血2ml送MMP-9檢測。2.2試驗儀器:MMP-9檢測所選用的設(shè)備主要有酶標儀、洗板機、數(shù)字顯示隔水式電熱恒溫培養(yǎng)箱、離心機、移液槍、漩渦混合器、烘箱、分液器及微量電動組織勻漿器等。2.3試驗過程:本實驗采用雙抗體夾心ABC-ELISA法。用抗人MMP-9單抗包被于酶標板上,標準品和樣品中的MMP-9與單抗結(jié)合,加入生物素化的抗人MMP-9,形成免疫復(fù)合物連接在板上,辣根過氧化物酶標記的Streptavidin與生物素結(jié)合,加入底物工作液顯藍色,最后加終止液硫酸,在450nm處測OD值,MMP-9濃度與OD值成正比,可通過繪制標準曲線求出標本中MMP-9濃度,將所獲得的數(shù)據(jù)再進行統(tǒng)計學(xué)分析。3、結(jié)果:3.1 MS(R-R型)患者其復(fù)發(fā)期和緩解期的MMP-9分別與正常對照組MMP-9比較均有統(tǒng)計學(xué)差異。3.2 頸椎病神經(jīng)根型患者其復(fù)發(fā)期和緩解期的MMP-9分別與正常對照組MMP-9比較沒有統(tǒng)計學(xué)差異。3.3 MS(R-R型)患者其復(fù)發(fā)期和緩解期的MMP-9分別與頸椎病神經(jīng)根型患者其復(fù)發(fā)期和緩解期的MMP-9比較均有統(tǒng)計學(xué)差異。3.4 MS患者的復(fù)發(fā)期與緩解期的MMP-9比較,有統(tǒng)計學(xué)差異。4、結(jié)論4.1 MMP-9的檢測對于MS(R-R型)和頸椎病神經(jīng)根型的鑒別具有臨床意義。4.2對于MS(R-R型)患者定期復(fù)查MMP-9可以早期判斷及預(yù)防有無疾病復(fù)發(fā),為積極診斷及治療提供幫助。4.3 MMP-9的檢測對于頸椎病神經(jīng)根型患者無現(xiàn)實臨床意義。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R744.51

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