Identification of Colon-specifi abnormal gene expression in alcerative colitis.

共进行了300次不同引物组合的展示。使用相同的引物组合，在至少2次展示后选择条带。regi β、IIIγ和碳酸酐酶I等基因在不同引物组的多次展示中被选择。我们成功克隆了118个条带。118个克隆中有68个（58%）被用作northern blotting探针，以确认GF、细菌重组和特异性无病原体（SPF）小鼠之间或小肠和结肠之间的差异基因表达。为了评估基因调控在细菌重构模型中的意义和特异性，我们建立了葡聚糖硫酸钠（DSS）诱导的结肠炎作为炎症对照。一些实验也使用了来自dss -结肠炎的rna。下面我们清楚地证明，我们的细菌重构模型和上皮基因筛选确实检测到与IBD相关的人类基因，这表明表1和表2中列出的一些基因一定与人类IBD有关。我们浓缩了两个分子。我们在IBD中证实了血清淀粉样蛋白A（SAA）的上皮诱导和SAA在脂多糖相关炎症中的调节活性。我们还发现再生基因III和HIP/PAP在实验和人类炎症性肠病中与Paneth细胞化生相关的上皮诱导。总之，我们通过分析细菌重组模型和研究人类同源物成功地鉴定了人类IBD的基因调控

A total of 300 displays with different sets of primers were carried out. Bands were selected after at least 2 displays using the same combination of primers. Some genes such as reg IIIβ, IIIγ and carbonic anhydrase I were selected in multiple displays with different sets of primers. We successfully cloned 118 bands. Sixty-eight clones out of 118 (58%) were used as probes in northern blotting to confirm differential gene expression among GF, bacteria-reconstituted and specific pathogen-free (SPF) mice, or between the small intestine and the colon. To evaluate the significance and specificity of gene modulation in the bacterial reconstitution model, we established dextran sulfate sodium (DSS) induced-colitis as an inflammatory control. RNAs from DSS-colitis were also used in some experiments.We clearly demonstrate in the following that our bacterial reconstitution model and epithelial gene screening truly detects human genes associated with IBD, suggesting that some genes listed in table 1 and 2 must be involved in human IBD.We concentrated two molecules. We demonstrated epithelial induction of serum amyloid A(SAA) in IBD and modulatory activity of SAA in LPS-associated inflammation. We additionally showed epithelial induction of Regenerating gene III and HIP/PAP in experimental and human inflammatory bowel disease in association with Paneth cell metaplasia.In summary, we successfully identified genes modulated in human IBD by analyzing bacteria reconstitution model and investigating human homologues