Enterococcus faecalis Metalloprotease Compromises Epithelial Barrier and Contributes to Intestinal Inflammation
Enterococcus faecalis Metalloprotease Compromises Epithelial Barrier and Contributes to Intestinal Inflammation
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DOI：
10.1053/j.gastro.2011.05.035
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发表时间：
2011-09-01
期刊：
影响因子：
29.4
通讯作者：
Haller, Dirk
中科院分区：
文献类型：
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作者：
Steck, Natalie;Hoffmann, Micha;Haller, Dirk
背景与目的：基质金属蛋白酶介导慢性肠道炎症的发病机制。我们描述了明胶酶（GelE），一种来自粪肠球菌的金属蛋白酶，在小鼠结肠炎发展中的作用。方法：无菌、白细胞介素-10缺陷（IL-10(-/-)）小鼠与大肠原性大肠杆菌菌株OG1RF和等基因、gele突变菌株单相关。通过测量IL-10（-/-）和TNF（δ ARE/Wt）小鼠结肠上皮细胞和结肠段中E-cadherin的表达、经上皮电阻（TER）和通透性标志物的易位来确定屏障功能。MMP抑制剂marimastat显示了GelE的特异性。结果：单粪肠球菌相关IL-10（-/-）小鼠的组织学分析（评分0-4）显示，在没有细菌来源的GelE的情况下，结肠组织炎症显著减少。我们在重组小鼠e -钙粘蛋白序列中发现了GelE的裂解位点，表明它可能被GelE降解。使用Ussing chambers和纯化的GelE进行的实验显示，在炎症发生前，肠炎症易感小鼠（IL-10（-/-）和TNF（δ ARE/Wt）小鼠）的屏障功能和细胞外E-cadherin的丧失。在用从克罗恩病和溃疡性结肠炎患者身上分离的OG1RF或粪肠杆菌菌株的GelE刺激后，结肠上皮细胞的TER减少，通透性标记物易位增加。结论：粪肠杆菌共生菌株产生的金属蛋白酶GelE通过破坏上皮屏障的完整性，促进了肠道炎症易感小鼠（IL-10（-/-）和TNF（δ are /Wt）小鼠）慢性肠道炎症的发展。
BACKGROUND & AIMS: Matrix metalloproteases (MMPs) mediate pathogenesis of chronic intestinal inflammation. We characterized the role of the gelatinase (GelE), a metalloprotease from Enterococcus faecalis, in the development of colitis in mice. METHODS: Germ-free, interleukin-10-deficient (IL-10(-/-)) mice were monoassociated with the colitogenic E faecalis strain OG1RF and isogenic, GelE-mutant strains. Barrier function was determined by measuring E-cadherin expression, transepithelial electrical resistance (TER), and translocation of permeability markers in colonic epithelial cells and colon segments from IL-10(-/-) and TNF(Delta ARE/Wt) mice. GelE specificity was shown with the MMP inhibitor marimastat. RESULTS: Histologic analysis (score 0-4) of E faecalis monoassociated IL-10(-/-) mice revealed a significant reduction in colonic tissue inflammation in the absence of bacteria-derived GelE. We identified cleavage sites for GelE in the sequence of recombinant mouse E-cadherin, indicating that it might be degraded by GelE. Experiments with Ussing chambers and purified GelE revealed the loss of barrier function and extracellular E-cadherin in mice susceptible to intestinal inflammation (IL-10(-/-) and TNF(Delta ARE/Wt) mice) before inflammation developed. Colonic epithelial cells had reduced TER and increased translocation of permeability markers after stimulation with GelE from OG1RF or strains of E faecalis isolated from patients with Crohn's disease and ulcerative colitis. CONCLUSIONS: The metalloprotease GelE, produced by commensal strains of E faecalis, contributes to development of chronic intestinal inflammation in mice that are susceptible to intestinal inflammation (IL-10(-/-) and TNF(Delta ARE/Wt) mice) by impairing epithelial barrier integrity.