Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution
Interleukin-13 is the key effector Th2 cytokine in ulcerative colitis that affects epithelial tight junctions, apoptosis, and cell restitution
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DOI：
10.1053/j.gastro.2005.05.002
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发表时间：
2005-08-01
期刊：
影响因子：
29.4
通讯作者：
Schulzke, JD
中科院分区：
文献类型：
--
作者：
Heller, F;Florian, P;Schulzke, JD
背景与目的：溃疡性结肠炎（UC）是一种以Th 2免疫应答为特征的疾病，伴有炎症反应和上皮屏障功能障碍。到目前为止，Th 2细胞因子尚未显示直接影响上皮屏障功能。（方法）酒吧下：刺激固有层单核细胞（LPMCs），用酶联免疫吸附测定法测定白细胞介素（IL）-13。在Ussing室中和通过电导扫描研究了IL-13和IL-4对HT 29/136结肠上皮细胞的功能性作用。末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记法检测细胞凋亡。通过逆转录聚合酶链反应和免疫荧光法分析IL-13/IL-4受体。采用Western印迹结合免疫荧光法检测紧密连接蛋白。此外，恢复速度进行了测量。最后，将UC患者的粘膜活检标本与培养细胞的这些特征进行比较。（结果）bar下：来自UC患者的LPMC产生大量的IL-13（985 +/- 73 pg/mL），比来自对照或克罗恩病患者的多得多。IL-13 R α 1和IL-4 R α受体存在于对照患者和UC患者的HT-29/B6细胞和结肠上皮细胞中。IL-13对HT-29/B6单层的跨上皮阻力具有剂量依赖性作用（降低至60% +/- 4%），而IL-4没有作用。这是由于凋亡细胞数量增加（5.6倍+/-0.9倍）和孔形成紧密连接蛋白claudin-2的表达增加至295% +/-37%，两者的贡献相等。最后，用IL-13处理后，上皮恢复速度从15.1 +/- 0.6 μ m/h降低到10.6 +/- 0.5 μ m/h。在人类样本中观察到平行变化，claudin-2表达增加至956% +/-252%。（结论）在酒吧之下：IL-13被鉴定为UC中的重要效应细胞因子，其通过影响上皮细胞凋亡、紧密连接和恢复速度来损害上皮屏障功能。
(Background& Aims:) under bar Ulcerative colitis (UC) is characterized by a Th2 immune response with inflammation and epithelial barrier dysfunction. So far, Th2 cytokines, have not been shown to directly influence epithelial barrier function. (Methods) under bar: Lamina propria mononuclear cells (LPMCs) were stimulated and interleukin (IL)-13 was measured by enzyme-linked immunosorbent assay. Functional IL-13 and IL-4 effects were studied on HT29/136 colonic epithelial cells in Ussing chambers and by conductance scanning. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assays. IL-13/IL-4 receptors were analyzed by reverse-transcription polymerase chain reaction and immunofluorescence. Western blotting combined with immunofluorescence was used to detect tight junction proteins. Furthermore, restitution velocity was measured. Finally, mucosal biopsy specimens from patients with UC were compared with cultured cells for these features. (Results) under bar: LPMCs from patients with UC produced large amounts of IL-13 (985 +/- 73 pg/mL), much more than from controls or patients with Crohn's disease. IL-13R alpha 1 and IL-4R alpha receptors were present in HT-29/B6 cells and colonic epithelial cells of control patients and patients with UC. IL-13 had a dose-dependent effect on transepithelial resistance of HT-29/B6 monolayers (reduction to 60% +/- 4%), whereas IL-4 had no effect. This was due to an increased number of apoptotic cells (5.6-fold +/- 0.9-fold) and an increased expression of the pore-forming tight junction protein claudin-2 to 295% +/- 37%, both of which contributed equally. Finally, epithelial restitution velocity decreased from 15.1 +/- 0.6 to 10.6 +/- 0.5 mu m/h after treatment with IL-13. Parallel changes were observed in human samples, with an increase in claudin-2 expression to 956% +/- 252%. (Conclusions) under bar: IL-13 was identified as an important effector cytokine in UC that impairs epithelial barrier function by affecting epithelial apoptosis, tight junctions, and restitution velocity.