Camilleri Michael, Carlson Paula, Valentin Nelson, Acosta Andres, O'Neill Jessica, Eckert Deborah, Dyer Roy, Na Jie, Klee Eric W, Murray Joseph A
Clinical Enteric Neuroscience Translational and Epidemiological Research (C.E.N.T.E.R.), Mayo Clinic, Rochester, Minnesota; and
Clinical Enteric Neuroscience Translational and Epidemiological Research (C.E.N.T.E.R.), Mayo Clinic, Rochester, Minnesota; and.
Am J Physiol Gastrointest Liver Physiol. 2016 Sep 1;311(3):G365-76. doi: 10.1152/ajpgi.00037.2016. Epub 2016 Jul 21.
Prior studies in with irritable bowel syndrome with diarrhea (IBS-D) patients showed immune activation, secretion, and barrier dysfunction in jejunal or colorectal mucosa. We measured mRNA expression by RT-PCR of 91 genes reflecting tight junction proteins, chemokines, innate immunity, ion channels, transmitters, housekeeping genes, and controls for DNA contamination and PCR efficiency in small intestinal mucosa from 15 IBS-D and 7 controls (biopsies negative for celiac disease). Fold change was calculated using 2((-ΔΔCT)) formula. Nominal P values (P < 0.05) were interpreted with false detection rate (FDR) correction (q value). Cluster analysis with Lens for Enrichment and Network Studies (LENS) explored connectivity of mechanisms. Upregulated genes (uncorrected P < 0.05) were related to ion transport (INADL, MAGI1, and SONS1), barrier (TJP1, 2, and 3 and CLDN) or immune functions (TLR3, IL15, and MAPKAPK5), or histamine metabolism (HNMT); downregulated genes were related to immune function (IL-1β, TGF-β1, and CCL20) or antigen detection (TLR1 and 8). The following genes were significantly upregulated (q < 0.05) in IBS-D: INADL, MAGI1, PPP2R5C, MAPKAPK5, TLR3, and IL-15. Among the 14 nominally upregulated genes, there was clustering of barrier and PDZ domains (TJP1, TJP2, TJP3, CLDN4, INADL, and MAGI1) and clustering of downregulated genes (CCL20, TLR1, IL1B, and TLR8). Protein expression of PPP2R5C in nuclear lysates was greater in patients with IBS-D and controls. There was increase in INADL protein (median 9.4 ng/ml) in patients with IBS-D relative to controls (median 5.8 ng/ml, P > 0.05). In conclusion, altered transcriptome (and to lesser extent protein) expression of ion transport, barrier, immune, and mast cell mechanisms in small bowel may reflect different alterations in function and deserves further study in IBS-D.
先前针对腹泻型肠易激综合征（IBS-D）患者的研究表明，空肠或结肠黏膜存在免疫激活、分泌及屏障功能障碍。我们通过逆转录聚合酶链反应（RT-PCR）检测了15例IBS-D患者和7例对照（乳糜泻活检阴性）小肠黏膜中91个反映紧密连接蛋白、趋化因子、固有免疫、离子通道、递质、管家基因以及用于DNA污染和PCR效率对照的基因的mRNA表达。使用2^((-ΔΔCT))公式计算倍数变化。名义P值（P < 0.05）经错误发现率（FDR）校正（q值）后进行解读。采用富集与网络研究透镜聚类分析（LENS）探索机制的连通性。上调基因（未校正P < 0.05）与离子转运（INADL、MAGI1和SONS1）、屏障（TJP1、2和3以及CLDN）或免疫功能（TLR3、IL15和MAPKAPK5）或组胺代谢（HNMT）相关；下调基因与免疫功能（IL-1β、TGF-β1和CCL20）或抗原检测（TLR1和8）相关。以下基因在IBS-D中显著上调（q < 0.05）：INADL、MAGI1、PPP2R5C、MAPKAPK5、TLR3和IL-15。在14个名义上上调的基因中，存在屏障和PDZ结构域的聚类（TJP1、TJP2、TJP3、CLDN4、INADL和MAGI1）以及下调基因的聚类（CCL20、TLR1、IL1B和TLR8）。IBS-D患者和对照的核裂解物中PPP2R5C的蛋白表达更高。IBS-D患者的INADL蛋白（中位数9.4 ng/ml）相对于对照（中位数5.8 ng/ml，P > 0.05）有所增加。总之，小肠中离子转运、屏障、免疫和肥大细胞机制的转录组（以及程度较轻的蛋白）表达改变可能反映了功能上不同的改变，值得在IBS-D中进一步研究。
