- 學位論文
明膠酶在黏液囊腫表現的研究
The study of the expression of gelatinases in mucoceles
摘要
基質金屬蛋白酶(matrix metalloproteinases，簡稱MMPs)是可以分解細胞外基質和基底膜的蛋白質水解酶（proteases），在許多疾病中扮演重要的角色。MMPs中的明膠酶(Gelatinases)，包括MMP-2及MMP-9，其主要作用是降解基底膜中的type IV膠原蛋白，在基底膜的破壞過程中扮演著重要的角色。由於先前研究曾指出黏液囊腫中的蛋白質水解酶與結締組織的重塑有相關，因此本研究擬探討黏液囊腫中的明膠酶的表現，進一步了解黏液囊腫的病理過程。本實驗的目的在於探索明膠酶在黏液囊腫中表達的情形。本實驗取30個黏液囊腫病例，施以蘇木精與依紅染色(hematoxylin and eosin stain，簡稱H＆E stain)和MMP-2、MMP-9免疫組織化學染色(immunohistochemistry stain，簡稱IHC染色)；每一個黏液囊腫標本皆製作一片蘇木精與依紅染色切片，作為判定黏液囊腫的發炎程度依據；分析黏液囊腫中MMP-2、MMP-9的表現程度與發炎程度的關連性，並以Fisher’s exact test和Spearman order rank correlation來檢定其是否具有統計學上的意義。此外，取黏液囊腫的組織與正常下唇小唾液腺的組織進行明膠酶譜(gelatin zymography)觀察，做MMP-2、MMP-9的半定量分析，並以Mann-Whitney Rank Sum Test方法統計。實驗結果顯示在黏液囊腫中幾乎沒有MMP-2的表達；但是黏液囊腫中可發現巨噬細胞、淋巴球、嗜中性球、血管內皮細胞等有MMP-9表達，且MMP-9的表現程度與黏液囊腫發炎程度呈現正相關(P<0.05)。另外明膠酶譜的結果，MMP-2的表現在黏液囊腫與正常唾液腺體中並沒有統計上的差異(P<0.05)；而MMP-9在黏液囊腫中有高度表現，但是在正常唾液腺體裡卻沒有(P<0.05)。本研究證實黏液囊腫中有MMP-9的存在，且進一步指出黏液囊腫的發炎程度與MMP-9表達程度有正相關。
並列摘要
Matrix metalloproteinases (MMPs), which degrade extracellular matrix (ECM) and basement membrane, plays an important role in many inflammatory diseases. Gelatinases including MMP-2 and MMP-9, which degrade type IV collagen, play an important role in the process of basement membrane degradation. Due to the relations between proteinases of mucocele and remodeling of connective tissue in past study, further study about the expression of gelatinases in mucoceles will help us understand the pathologic process of mucocele formation. The aim of this study was to investigate the expression of gelatinases alteration in mucoceles. Thirty specimens were used in this study. All specimens were studied both histologically and MMP-2, MMP-9 immunohistochemistry stain. One of each tissue blocks was sliced and stained with hematoxylin and eosin for histological diagnosis of inflammation degree. The correlation between degree of inflammation and activity of MMP-9 was examined by the Fisher’s exact test and Spearman rank order correlation. Tissues include 12 mucoceles and 6 normal small saliva glands over lower lip were used to evaluate MMP-2 and MMP-9 activities by gelatin zymography. Bands on the zymograms were quantified by using the AlphaImager 2000 densitometer and statistical analysis was carried out by Mann-Whitney Rank Sum Test. These results manifest the expression of MMP-9 but not MMP-2 in mucoceles. After further statistical analysis, we found that the correlation between degree of inflammation and the expression of MMP-9 in mucoceles was statistically significant (P<0.05). Gelatin zymography display that MMP-9 highly expression in mucoceles but none on normal saliva glands(P<0.05). The study demonstrates that MMP-9 was exist in mucocele and the expression was related to degree of inflammation.