腎臟間質纖維化與基質金屬蛋白酶及基質金屬蛋白酶抑制劑之相關性

The Association between Renal Interstitial Fibrosis and Matrix Metalloproteinases and Tissue Inhibitor of Metalloproteinases

高維澤(Wei-Tse Kao)

指導教授 ： 張浤榮

中山醫學大學/醫學院/醫學研究所/碩士(2012年)

https://doi.org/10.6834/CSMU.2012.00027

摘要

腎臟纖維化是大多數慢性腎病變進展到末期腎病的共同表現，而腎臟纖維化的病理機轉特點則為腎臟組織細胞外基質成份的過度累積與沈積。基質金屬蛋白酶（matrix metalloproteinases；MMPs）及基質金屬蛋白酶抑制劑（tissue inhibitor of metalloproteinases；TIMPs）被認為與腎臟細胞外基質的重組化有關連。先前的研究大都僅限於動物實驗或是人體血清的MMPs/TIMPs活性與腎臟纖維化的關係，因此本研究的目標是進一步探討MMPs/TIMPs的活性與腎臟纖維化在組織的相關性。
46位接受腎切除手術的患者被納入腎臟間質纖維化研究，並依照其腎臟組織間質纖維化程度（interstitial fibrosis score ; IFS）而分為實驗組（high-grade interstitial fibrosis group）及對照組（low-grade interstitial fibrosis group）。分析其不包含腫瘤及病灶的腎臟組織的MMP-9、MMP-2及TIMP-1免疫組織化學染色之表現，這項研究的目標是比較這兩組病人中，相關臨床數據與MMP-9、MMP-2及TIMP-1的免疫組織化學染色的表現與腎間質纖維化的相關性。
結果發現腎臟間質纖維化程度較高者在萎縮腎小管細胞質及腎絲球細胞質都有MMP-9的免疫組織化學染色表現比率顯著地降低，在萎縮腎小管細胞質之實驗組比對照組為：55.2%比94.1%（p=0.007）；腎絲球細胞質之實驗組比對照組：62.1%比100%（p=0.003）。此外也發現腎臟間質纖維化較高者也都在萎縮腎小管細胞質有MMP-2的免疫組織化學染色表現比率顯著地增加（實驗組比對照組：55.2%比5.9%，p=0.034）以及TIMP-1的免疫組織化學染色表現比率顯著地增加（實驗組比對照組：100%比64.7%，p=0.001）的情形。
本研究則發現腎臟間質纖維化較高者在萎縮腎小管細胞質有MMP-9的免疫組織化學染色表現比率顯著地降低、MMP-2的免疫組織化學染色表現比率顯著地增加以及TIMP-1的免疫組織化學染色表現比率顯著地增加。單變項分析中顯示腎間質纖維化程度與下列有正相關：合併有慢性腎病、血清肌酐酸、MMP-9在萎縮腎小管的細胞核上之表現、MMP-2在萎縮腎小管的細胞質上之表現以及TIMP-1在萎縮腎小管的細胞質上之表現（p值分別為p=0.020, p<0.001, p=0.004, p=0.021及p=0.039）。此外單變項分析中顯示腎間質纖維化程度與下列有負相關：估計的腎絲球過濾率、MMP-9在正常腎小管的細胞質上之表現、MMP-9在腎絲球的細胞質上之表現、MMP-9在萎縮腎小管的細胞質上之表現以及TIMP-1在腎絲球的細胞核上之表現（p值分別為p<0.001, p<0.001, p=0.012, p<0.001及p=0.028）。在逐步多變項回歸分析法中，則發現估計的腎絲球過濾率以及MMP-9在正常腎小管細胞質內的免疫組織化學染色表現是最主要的預測腎間質纖維化之因素（p=0.001以及p<0.001)。
結論是：人類腎間質纖維化與估計的腎絲球過濾率以及MMP-9在正常腎小管的細胞質上之免疫組織化學表現的活性減少有相關性。

關鍵字

慢性腎病 ； 基質金屬蛋白酶免疫組織化學染色 ； 腎間質纖維化

並列摘要

Introduction Renal fibrosis is the final common manifestation of almost all forms of chronic kidney disease (CKD) that progress to end-stage renal failure. The pathogenesis of renal fibrosis is the process characterized by an excessive accumulation and deposition of extracellular matrix components (ECM). It has been suggested that matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) might involve in the remodeling of the ECM. Previous animal studies had showed the correlation of renal fibrosis and the MMPs/TIMPs expressions but there were only limited human studies that revealed the relation of the renal fibrosis and the activity of serum MMPs/TIMPs. We intended to study the roles of MMP-9, MMP-2 and TIMP-1 in the pathogenesis of human renal interstitial fibrosis.
Materials and Methods Forty-six patients who received nephrectomy were retrospectively enrolled and they were divided into low-grade and high-grade interstitial fibrosis groups according the interstitial fibrosis scores. The immunohistochemical (IHC) expressions of MMP-9, MMP-2 and TIMP-1 in the non-tumor parts of these kidneys were examined and the correlations of IHC expressions of MMP-9, MMP-2 and TIMP-1 between the low-grade and high-grade interstitial fibrosis groups were analyzed.
Results Patients with high-grade interstitial fibrosis had significantly lower MMP-9 IHC expression percentage over atrophic tubular cytoplasm (high-grade vs. low-grade: 55.2% vs. 94.1%, p=0.007) and glomerular cytoplasm (high-grade vs. low-grade: 62.1% vs. 100%, p=0.003). And patients with high-grade interstitial fibrosis had significantly higher MMP-2 and TIMP-1 IHC expression percentage over atrophic tubular cytoplasm (MMP-2: high-grade vs. low-grade: 55.2% vs. 5.9%, p=0.034; TIMP-1: high-grade vs. low-grade: 100% vs. 64.7%, p=0.001). Univariate analysis showed that interstitial fibrosis correlated positively with percentage of CKD (p=0.020), serum creatinine level (p<0.001), MMP-9 atrophic tubular nuclei expression (p=0.004), MMP-2 atrophic tubular cytoplasm expression (p=0.021) and TIMP-1 atrophic tubular cytoplasm expression (p=0.039) and negatively with estimate glomerular filtration rate (p<0.001), MMP-9 normal and atrophic tubular cytoplasm expression (both p<0.001) and MMP-9 glomerular cytoplasm expression (p=0.012). Stepwise multivariate regression revealed that estimate glomerular filtration rate (eGFR) and MMP-9 expression over normal tubular cytoplasm (p=0.001; p<0.001, respectively) were the most two significant factors that predicted renal interstitial fibrosis score.
Conclusion Renal interstitial fibrosis was negatively associated with eGFR and MMP-9 expression over normal tubular cytoplasm.

並列關鍵字

chronic kidney disease ； matrix metalloproteinases ； immunohistochemical stain ； renal interstitial fibrosis

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