A murine model of chronic inflammation-induced intestinal fibrosis down-regulated by antisense NF-κB
A murine model of chronic inflammation-induced intestinal fibrosis down-regulated by antisense NF-κB
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DOI：
10.1053/j.gastro.2003.08.027
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发表时间：
2003-12-01
期刊：
影响因子：
29.4
通讯作者：
Chakravarti, S
中科院分区：
文献类型：
--
作者：
Lawrance, IC;Wu, F;Chakravarti, S
背景和目标：为了阐明肠纤维化（炎症性肠病的常见并发症）的细胞外基质（ECM）变化，我们建立了与肠纤维化相关的慢性结肠炎小鼠模型。方法：采用三硝基苯磺酸（TNBS）每周一次直肠内给药建立慢性炎症模型。在该模型的2个变体中，给予核因子K B（NF-κ B）p65的反义寡核苷酸治疗性或治疗性以阻断慢性炎症相关的纤维化。通过组织学确定结肠炎症和纤维化。通过羟脯氨酸定量来估计总胶原水平。结肠胶原表达（Col1a2，Col3a2），ECM重塑基因（基质金属蛋白酶[MMP]-1、-3和基质金属蛋白酶组织抑制剂[TIMP]-1）和炎症调节细胞因子（肿瘤坏死因子α [TNF-α]，干扰素γ [IFN-γ]，转化生长因子β 1 [TGF-β 1]，和胰岛素样生长因子1 [IGF-1]）通过半定量逆转录聚合酶链反应进行评估。对照和TNBS处理的结肠间充质细胞的特征在于形态，表型和功能的TNF-α和IFN-γ的反应。结果如下：TNBS治疗小鼠的结肠包含急性和慢性炎性浸润、胶原蛋白增加、纤维化组织结构以及TNF-α、TGF-β 1、IGF-1、Col1a2、MMP-1和TIMP-1表达增加。TNBS处理小鼠的结肠间充质细胞在形态学上也与对照小鼠不同，IFN-γ处理后TIMP-1表达增加。在TNBS治疗停止后，纤维化持续2 - 4周。在给予NF-kappaB反义寡核苷酸的小鼠中，67%没有纤维化，而在建立慢性炎症后治疗的小鼠中，43%没有纤维化。结论：小鼠的长期TNBS治疗产生了慢性肠道炎症相关的纤维化，并伴有广泛的纤维化ECM变化，这些变化可以通过特异性阻断NF-κ B来抵消。
Background & Aims: To elucidate extracellular matrix (ECM) changes underlying intestinal fibrosis, a frequent complication of inflammatory bowel disease, we developed a murine model of chronic colitis associated with intestinal fibrosis. Methods: Chronic inflammation was established by weekly intrarectal administration of trinitrobenzene sulfonic acid (TNBS). In 2 variations of the model an antisense oligonucleotide for nuclear factor K B (NF-kappaB) p65 was given prophylactically or therapeutically to block chronic inflammation-associated fibrosis. Colonic inflammation and fibrosis were determined by histology. Total collagen level was estimated by hydroxyproline quantification. Colonic expression of collagens (Col1a2, Col3a2), ECM remodeling genes (matrix metalloproteinase [MMP]-1, -3, and tissue inhibitor of matrix metalloproteinase [TIMP]-1), and inflammation-modulating cytokines (tumor necrosis factor alpha [TNF-alpha], interferon gamma [IFN-gamma], transforming growth factor beta1 [TGF-beta1], and insulin-like growth factor 1 [IGF-1]) were assessed by semiquantitative reverse-transcription polymerase chain reaction. Control and TNBS-treated colonic mesenchymal cells were characterized by morphology, phenotype, and functional response to TNF-alpha and IFN-gamma. Results: Colons of TNBS-treated mice contained acute and chronic inflammatory infiltrates, increased collagen, fibrogenic tissue architecture, and increased expression of TNF-alpha, TGF-beta1, IGF-1, Col1a2, MMP-1, and TIMP-1. Colonic mesenchymal cells from TNBS-treated mice were also morphologically distinct from those of the control mice, with increased TIMP-1 expression in response to IFN-gamma treatment. Fibrosis persisted for 2-4 weeks after cessation of the TNBS treatment. In mice given NF-kappaB antisense prophylactically, 67% were fibrosis-free, whereas of those treated after establishing chronic inflammation, 43% were free of fibrosis. Conclusions: Extended TNBS treatment of mice yielded chronic intestinal inflammation-associated fibrosis with extensive fibrogenic ECM changes that could be counteracted by specific blockade of NF-kappaB.