Functional modulation of crohn's disease myofibroblasts by anti-tumor necrosis factor antibodies
Functional modulation of crohn's disease myofibroblasts by anti-tumor necrosis factor antibodies
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DOI：
10.1053/j.gastro.2007.04.069
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发表时间：
2007-07-01
期刊：
影响因子：
29.4
通讯作者：
Macdonald, Thomas T.
中科院分区：
文献类型：
--
作者：
Di Sabatino, Antonio;Pender, Sylvia L. F.;Macdonald, Thomas T.
背景和目的：英夫利昔单抗通过跨膜肿瘤坏死因子-α (mTNF) 从外到内的信号传导诱导免疫细胞凋亡。然而，在发炎的肠道中，肌成纤维细胞也会产生 TNF-α，而抗 TNF 抗体对这些结构细胞的影响尚不清楚。我们研究了英夫利昔单抗对细胞凋亡、基质金属蛋白酶 (MMP) 和金属蛋白酶组织抑制剂 (TIMP)-1 的产生以及克罗恩病 (CD) 肌成纤维细胞迁移的作用。方法：从活动性 CD 患者和对照组中分离结肠肌成纤维细胞。通过蛋白质印迹和流式细胞术评估 mTNF。通过膜联蛋白 V 染色和 caspase-3 分析英夫利昔单抗处理的肌成纤维细胞的细胞凋亡。通过蛋白质印迹法测量 TIMP-1 和 MMP，并使用体外伤口愈合划痕试验评估成纤维细胞迁移。结果：CD 肌成纤维细胞表现出比对照肌成纤维细胞更高的 mTNF 表达。英夫利昔单抗对 CD 肌成纤维细胞凋亡、caspase-3 激活以及 MMP-3 和 MMP-12 的产生没有影响。然而，英夫利昔单抗诱导 TIMP-1 产生显着的剂量依赖性增加，这种增加被 p38 丝裂原激活蛋白激酶抑制剂 SB 203580 抑制。抗 TNF 药物阿达木单抗、依那西普和 p55 TNF 受体-人 IgG 融合蛋白也增加 TIMP-1 产生。英夫利昔单抗和重组人TIMP-1显着增强CD肌成纤维细胞的迁移，而英夫利昔单抗诱导的迁移被α-TIMP-1中和抗体抑制。英夫利昔单抗还减少 CD 肌成纤维细胞胶原蛋白的产生。结论：我们的研究结果显示了抗 TNF 疗法在增强 TIMP-1 产生和肌成纤维细胞迁移方面的新治疗途径，这可能会降低 MMP 活性并促进伤口愈合。
Background & Aims: Infliximab induces immune cell apoptosis by outside-to-inside signaling through transmembrane tumor necrosis factor-alpha (mTNF). However, in inflamed gut, myofibroblasts also produce TNF-alpha, and the affects of anti-TNF antibodies on these structural cells are unknown. We investigated the action of infliximab on apoptosis, the production of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMP)-1, and migration of Crohn's disease (CD) myofibroblasts. Methods: Colonic myofibroblasts were isolated from patients with active CD and controls. mTNF was evaluated by Western blotting and flow cytometry. infliximab-treated myofibroblasts were analyzed for apoptosis by Annexin V staining and caspase-3. TIMP-1 and MMPs were measured by Western blotting, and fibroblast migration was assessed by using an in vitro wound-healing scratch assay. Results: CD myofibroblasts showed higher mTNF expression than control myofibroblasts. Infliximab had no effect on CD myofibroblast apoptosis, caspase-3 activation, and production of MMP-3 and MMP-12. However, infliximab induced a significant dose-dependent increase in TIMP-1 production, which was inhibited by the p38 mitogen-activated protein kinase inhibitor SB 203580. The anti-TNF agents adalimumab, etanercept, and p55 TNF-receptor-human IgG fusion protein also increased TIMP-1 production. The migration of CD myofibroblasts was enhanced significantly by infliximab and recombinant human TIMP-1, and infliximab-induced migration was inhibited by and-TIMP-1 neutralizing antibody. Infliximab also decreased CD myofibroblast collagen production. Conclusions: Our findings show a novel therapeutic pathway for anti-TNF therapies in enhancing TIMP-1 production and myofibroblast migration, which may reduce MMP activity and facilitate the wound healing.