【摘要】：目的:通過實(shí)驗(yàn)研究探討人退變腰椎間盤髓核組織中MMP-28的表達(dá)及變化情況,從而評(píng)價(jià)在人腰椎間盤退變疾病過程中MMP-28所扮演的角色及作用,并分析退變等級(jí)與其關(guān)聯(lián)性。方法:所有患者術(shù)前均行腰椎MRI等檢測(cè),根據(jù)影像學(xué)資料及術(shù)中所見將腰椎間盤退變分為:正常組、突出組、脫出組及游離組。實(shí)驗(yàn)通過蘇木精-伊紅染色法(hematoxylin-eosin staining HE)去觀察正常椎間盤組織及退變椎間盤組織的形態(tài)差異,通過免疫組織化學(xué)技術(shù)(Immunohis to chemistry IHC)檢測(cè)觀察MMP-28處于腰椎間盤髓核組織中的部位,使用Western-blot檢測(cè)MMP-28在各個(gè)等級(jí)退變椎間盤中的蛋白表達(dá)水平。全部標(biāo)本均按照選取標(biāo)準(zhǔn)取材,且所取組織標(biāo)本均來源于腰椎間盤纖維環(huán)內(nèi)的髓核,盡可能的避免及排除椎管內(nèi)其余物質(zhì)對(duì)髓核標(biāo)本組織的影響,使實(shí)驗(yàn)更能夠客觀真實(shí)地反映髓核中MMP-28的表達(dá)及變化狀況,實(shí)驗(yàn)結(jié)果數(shù)據(jù)更加可靠且具有說服力。P0.05具有統(tǒng)計(jì)學(xué)上顯著差異性。結(jié)果:蘇木精-伊紅染色法結(jié)果顯示正常組及退變組的細(xì)胞數(shù)目及形態(tài)均有明顯差異。免疫組化結(jié)果顯示MMP-28在腰椎間盤髓核細(xì)胞中的定位為胞漿內(nèi),在正常組及退變組中均有表達(dá),但退變組細(xì)胞染色數(shù)量及程度均較正常組明顯。Western-blot結(jié)果顯示MMP-28表達(dá)于各等級(jí)椎間盤組織中,且與椎間盤退變等級(jí)存在一定相關(guān)性,各等級(jí)之間均具有差異且有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:人類腰椎間盤髓核組織的退變過程中有MMP-28的參與,且與其表達(dá)存在關(guān)聯(lián)性,同時(shí)MMP-28隨著人類退變腰椎間盤組織退變等級(jí)的加重具有相同的增高趨勢(shì)。
[Abstract]:Objective: to investigate the expression and changes of MMP-28 in the nucleus pulposus of human degenerative lumbar intervertebral disc, and to evaluate the role and role of MMP-28 in the process of human lumbar disc degeneration. The relationship between the grade of degeneration and its relationship is also analyzed. Methods: all patients were examined for lumbar vertebra MRI before operation. According to imaging data and intraoperative findings, lumbar intervertebral disc degeneration was divided into normal group, prolapse group and free group. The morphological differences between normal intervertebral disc tissue and degenerative intervertebral disc tissue were observed by hematoxylin-eosin staining (hematoxylin-eosin staining HE). Immunohistochemical technique (Immunohis to chemistry IHC) was used to detect the location of MMP-28 in the nucleus pulposus of lumbar intervertebral disc, and Western-blot was used to detect the protein expression level of MMP-28 in various grades of degenerative intervertebral disc. All the specimens were taken according to the selection criteria, and the tissue samples were derived from the nucleus pulposus in the lumbar intervertebral disc annulus, so as to avoid and exclude the influence of other substances in the spinal canal on the tissue of the nucleus pulposus specimen as far as possible. The experiment can reflect the expression and change of MMP-28 in nucleus pulposus objectively and truthfully. The experimental data are more reliable and persuasive. P0.05 has statistically significant difference. Results: the results of hematoxylin-eosin staining showed that there were significant differences in cell number and morphology between normal group and degenerative group. Immunohistochemical results showed that MMP-28 was localized in cytoplasm in nucleus pulposus cells of lumbar intervertebral disc, and was expressed in normal and degenerative groups. The results of Western-blot showed that MMP-28 was expressed in all grades of intervertebral disc tissue, and there was a certain correlation between the expression of MMP-28 and the grade of degeneration of intervertebral disc, but the number and degree of staining of cells in the degeneration group were more obvious than those in the normal group. There were significant differences among different grades (P0.05). Conclusion: MMP-28 is involved in the degeneration of human lumbar disc nucleus pulposus, and it is associated with its expression. Meanwhile, MMP-28 has the same increasing trend with the aggravation of degeneration grade of human lumbar intervertebral disc tissue.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R681.53

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