Downregulation of epithelial apoptosis and barrier repair in active Crohn's disease by tumour necrosis factor α antibody treatment
Downregulation of epithelial apoptosis and barrier repair in active Crohn's disease by tumour necrosis factor α antibody treatment
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DOI：
10.1136/gut.2003.036632
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发表时间：
2004-09-01
期刊：
影响因子：
24.5
通讯作者：
Schulzke, JD
中科院分区：
文献类型：
--
作者：
Zeissig, S;Bojarski, C;Schulzke, JD
背景和目的：屏障功能障碍是导致克罗恩病（CD）炎症和腹泻的重要特征。最近，肿瘤坏死因子α (tnf - α)抗体被认为对类固醇难治性CD有效。本研究的目的是表征这种治疗对上皮屏障的影响。患者和方法：11例慢性活动性CD（克罗恩病活动性指数bbbb150）患者，在tnf - α抗体治疗前和治疗后14天，从乙状结肠取钳活检。采用末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口端标记（TUNEL）和4′，6-二氨基-2-苯基吲哚染色检测上皮细胞凋亡。上皮电阻在小型化的乌辛腔中用交流阻抗分析法测定。免疫印迹法定量测定Occludin、claudin 1和claudin 4的表达。结果：对照组上皮细胞凋亡率为2.1 (0.2)%，CD组细胞凋亡率为5.3(1.0)%。tnf - α抗体治疗使细胞凋亡率降至2.9(1.0)%（11例患者中有10例恢复正常）。同时，与对照组相比，CD组上皮细胞的耐药性较低(24 (3)vs 42 (3) Omegaxcm(2))，治疗后改善至34 (3)Omegaxcm(2)。tnf -抗体治疗不影响Occludin、claudin 1和claudin 4。为了支持上皮细胞凋亡在CD中的功能作用，在HT-29/B6细胞中，喜树碱选择性地将凋亡率从2.6%上调至5.4%时，观察到类似的-40%的抗性下降。结论：11例患者中有10例经tnf - α抗体治疗后，结肠上皮细胞凋亡水平上调，恢复正常。这是上皮屏障功能障碍的结构相关性，被测量为上皮抗性，而紧密连接蛋白的表达对这种治疗效果没有贡献。
Background and aims: Barrier dysfunction is an important feature contributing to inflammation and diarrhoea in Crohn's disease (CD). Recently, tumour necrosis factor alpha (TNF-alpha) antibodies were recognised as effective in steroid refractory CD. The aim of this study was to characterise the effects of this therapy on the epithelial barrier.Patients and methods: Forceps biopsies were obtained from the sigmoid colon before and 14 days after TNF-alpha antibody therapy in 11 patients treated for chronic active CD (Crohn's disease activity index >150). Epithelial apoptoses were measured after terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling (TUNEL) and 4',6-diamidino-2-phenylindole staining. Epithelial resistance was determined by alternating current impedance analysis in miniaturised Ussing chambers. Occludin, claudin 1, and claudin 4 expression was quantified in immunoblots.Results: The epithelial apoptotic ratio was 2.1 (0.2)% in controls and increased to 5.3 (1.0)% in CD. TNF-alpha antibody therapy decreased the apoptotic ratio to 2.9 (1.0)% (normalised in 10 of 11 patients). In parallel, epithelial resistance was lower in CD than in controls (24 (3) v 42 (3) Omegaxcm(2)) and improved to 34 (3) Omegaxcm(2) after therapy. Occludin, claudin 1, and claudin 4 were not affected by TNF-alpha antibody therapy. In support of a functional role of epithelial apoptoses in CD, a similar decrease in resistance of -40% was observed when the apoptotic rate was selectively upregulated from 2.6% to 5.4% with camptothecin in HT-29/B6 cells.Conclusions: Epithelial apoptoses were upregulated in the colon in CD and restored to normal in 10 of 11 patients by TNF-alpha antibody therapy. This is the structural correlate of epithelial barrier dysfunction measured as epithelial resistance while expression of tight junction proteins did not contribute to this therapeutic effect.