Stress-induced disruption of colonic epithelial barrier:: Role of interferon-γ and myosin light chain kinase in mice
Stress-induced disruption of colonic epithelial barrier:: Role of interferon-γ and myosin light chain kinase in mice
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DOI：
10.1016/s0016-5085(03)01057-6
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发表时间：
2003-09-01
期刊：
影响因子：
29.4
通讯作者：
Bueno, L
中科院分区：
文献类型：
--
作者：
Ferrier, L;Mazelin, L;Bueno, L
背景和目标：应激性生活事件被认为与炎症性肠病和肠易激综合征等多种疾病有关。肠上皮屏障功能受损是应激的一个可疑结果，但其潜在机制尚不清楚。本研究旨在确定应激调节结肠上皮屏障的机制。研究方法：每天2小时应激1-4天后，通过竞争性逆转录聚合酶链反应评价小鼠结肠、肝脏和脾脏中细胞因子信使RNA（mRNA）的表达。结肠细胞旁通透性测定为Cr-51-乙二胺四乙酸的体内管腔-血液比。肌球蛋白轻链（MLC）激酶抑制剂（ML-7）对应激诱导的干扰素（IFN）-γ mRNA表达和结肠上皮屏障损伤的影响进行了评估，并通过免疫印迹法测定MLC磷酸化。最后，确定了反复应激对细菌移位的影响。结果：反复应激诱导结肠IFN-γ过度表达。在肝脏中，检测到更高水平的IFN-γ、白细胞介素（IL）-4和IL-10 mRNA，并与细菌移位、炎症和细胞凋亡相关。应激增加了对照组小鼠的结肠通透性，但对SCID和IFN-γ缺陷小鼠没有影响。ML-7抑制了应激诱导的肝脏通透性增加、细菌移位和细胞因子过表达，并恢复了正常的组织学。在应激动物中检测到更大量的磷酸化MLC。结论：重复的应激过程驱动器官特异性细胞因子表达模式，并改变与细菌移位相关的结肠粘膜屏障功能。这种效应依赖于CD 4（+）T细胞的存在，并需要IFN-γ的产生和MLC磷酸化。
Background & Aims: Stressful life events are supposed to be involved in various diseases such as inflammatory bowel diseases and irritable bowel syndrome. Impairment of the intestinal epithelial barrier function is a suspected consequence of stress, but the underlying mechanisms remain unclear. This study aimed to determine the mechanisms through which stress modulates the colonic epithelial barrier. Methods: Cytokine messenger RNA (mRNA) expression was evaluated in murine colon, liver, and spleen by competitive reverse-transcription polymerase chain reaction after 1-4 days of daily 2-hour stress sessions. Colonic paracellular permeability was measured as the in vivo lumen-to-blood ratio of Cr-51-ethylenediaminetetraacetic acid. The effect of a myosin light chain (MLC) kinase inhibitor (ML-7) was assessed on stress-induced interferon (IFN)-gamma mRNA expression and colonic epithelial barrier impairment, and MLC phosphorylation was determined by immunoblot. Finally, the incidence of repeated stress sessions on bacterial translocation was determined. Results: Repeated stress induced an overexpression of colonic IFN-gamma. In the liver, higher levels of IFN-gamma, interleukin (IL)-4, and IL-10 mRNAs were detected and were associated with bacterial translocation, inflammation, and apoptosis. Stress increased colonic permeability of control mice, but not of SCID and IFN-gamma-deficient mice. ML-7 inhibited the stress-induced increased permeability, bacterial translocation, and cytokine overexpression in the liver and restored a normal histology. Larger amounts of phosphorylated MLC were detected in stressed animals. Conclusions: Repeated stress sessions drive organ-specific cytokine expression patterns and alter colonic mucosal barrier functions associated with bacterial translocation. This effect depends on the presence of CD4(+) T cells and requires IFN-gamma production and MLC phosphorylation.