Zheng Bin, Morgan Mary E, van de Kant Hendrik J G, Garssen Johan, Folkerts Gert, Kraneveld Aletta D
Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, The Netherlands.
Biochim Biophys Acta. 2013 Dec;1832(12):2162-72. doi: 10.1016/j.bbadis.2013.07.004. Epub 2013 Jul 12.
Pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), contribute to the development of intestinal inflammatory diseases, like inflammatory bowel disease (IBD). Supporting investigations of the underlying mechanisms of IBD, this study provides an extensive PRR expression survey together with T-cell associated factors along the murine colon during experimental colitis. 8-12 week-old C57BL/6 mice were treated with dextran sodium sulfate (DSS) to induce colitis. The mRNA expression levels of Tlr1-9, Nod1, Nod2, T cell subset-associated master transcription factors and cytokines were determined using qPCR. The expression of TLR2, 4, 5 and 6 was determined with immunohistochemistry. Th1 and Th17 associated responses were quantified in the mesenteric lymph nodes (mLNs) using flow cytometry. In DSS treated mice, the mRNA expression of the majority of PRRs was increased relative to healthy controls and correlated with the degree of inflammation. The exceptions were Tlr1 and Tlr5, which displayed unchanged and down-regulated transcription, respectively. Furthermore, in healthy animals, there was increased transcription of Tlr2, 3 and 5 near the caecum as opposed the region near the rectum. Within the inflamed regions, the mRNA expression of Th1-, Th17- and regulatory T-cell associated cytokines was enhanced, while there was no change for Th2-associated cytokines. In agreement with the mRNA expression, enhanced IFNγ and IL-17 producing cells were observed in stimulated mLNs. This study provides an extensive expression survey of PRRs along the colon during the acute colitis and shows that the induced inflammation is characterized by a Th1- and IL-17 mediated cytokine response.
模式识别受体（PRR），如Toll样受体（TLR），在诸如炎症性肠病（IBD）等肠道炎症性疾病的发展中起作用。为支持对IBD潜在机制的研究，本研究对实验性结肠炎期间小鼠结肠中的PRR表达以及T细胞相关因子进行了广泛调查。用葡聚糖硫酸钠（DSS）处理8 - 12周龄的C57BL / 6小鼠以诱导结肠炎。使用qPCR测定Tlr1 - 9、Nod1、Nod2、T细胞亚群相关主转录因子和细胞因子的mRNA表达水平。用免疫组织化学法测定TLR2、4、5和6的表达。使用流式细胞术对肠系膜淋巴结（mLN）中的Th1和Th17相关反应进行定量。在DSS处理的小鼠中，大多数PRR的mRNA表达相对于健康对照增加，并且与炎症程度相关。例外情况是Tlr1和Tlr5，其转录分别未改变和下调。此外，在健康动物中，与直肠附近区域相反，盲肠附近的Tlr2、3和5转录增加。在炎症区域内，Th1、Th17和调节性T细胞相关细胞因子的mRNA表达增强，而Th2相关细胞因子没有变化。与mRNA表达一致，在受刺激的mLN中观察到产生IFNγ和IL - 17的细胞增加。本研究对急性结肠炎期间结肠中的PRR进行了广泛的表达调查，并表明诱导的炎症以Th1和IL - 17介导的细胞因子反应为特征。
