Clinical Enteric Neuroscience Translational and Epidemiological Research (C.E.N.T.E.R.) and
Clinical Enteric Neuroscience Translational and Epidemiological Research (C.E.N.T.E.R.) and.
Am J Physiol Gastrointest Liver Physiol. 2014 Jun 15;306(12):G1089-98. doi: 10.1152/ajpgi.00068.2014. Epub 2014 Apr 24.
Our aim was to conduct a pilot case-control study of RNA expression profile using RNA sequencing of rectosigmoid mucosa of nine females with -diarrhea-predominant irritable bowel syndrome (IBS-D) with accelerated colonic transit and nine female healthy controls. Mucosal total RNA was isolated and purified, and next-generation pair-end sequencing was performed using Illumina TruSeq. Analysis was carried out using a targeted approach toward 12 genes previously associated with IBS and a hypothesis-generating approach. Of the 12 targeted genes tested, patients with IBS-D had decreased mRNA expression of TNFSF15 (fold change controls to IBS-D: 1.53, P = 0.01). Overall, up- and downregulated mRNA expressions of 21 genes (P = 10(-5) to 10(-8); P values with false detection rates are shown) were potentially relevant to IBS-D including the following: neurotransmitters [P2RY4 (P = 0.001), vasoactive intestinal peptide (VIP, P = 0.02)]; cytokines [CCL20 (P = 0.019)]; immune function [C4BPA complement cascade (P = 0.0187)]; interferon-related [IFIT3 (P = 0.016)]; mucosal repair and cell adhesion [trefoil protein (TFF1, P = 0.012)], retinol binding protein [RBP2 (P = 0.017)]; fibronectin (FN1, P = 0.009); and ion channel functions [guanylate cyclase (GUCA2B, P = 0.017), PDZ domain-containing protein 3 (PDZD3, P = 0.029)]. Ten genes associated with functions related to pathobiology of IBS-D were validated by RT-PCR. There was significant correlation in fold changes of the selected genes (Rs = 0.73, P = 0.013). Up- or downregulation of P2RY4, GUC2AB, RBP2, FNI, and C4BPA genes were confirmed on RT-PCR, which also revealed upregulation of farnesoid X receptor (FXR) and apical sodium-coupled bile acid transporter (IBAT/ASBT). RNA-Seq and RT-PCR analysis of rectosigmoid mucosa in IBS-D show transcriptome changes that provide the rationale for validation studies to explore the role of mucosal factors in the pathobiology of IBS-D.
我们的目的是使用九名女性直肠乙状结肠黏膜的 RNA 测序进行 RNA 表达谱的初步病例对照研究，这些女性患有腹泻为主的肠易激综合征（IBS-D），伴有结肠转运加速，九名女性健康对照。分离并纯化黏膜总 RNA，然后使用 Illumina TruSeq 进行下一代端对端测序。使用针对先前与 IBS 相关的 12 个基因的靶向方法和生成假设的方法进行分析。在测试的 12 个靶向基因中，IBS-D 患者的 TNFSF15 mRNA 表达降低（对照到 IBS-D 的折叠变化：1.53，P=0.01）。总体而言，21 个基因的 mRNA 表达上调和下调（P=10（-5）至 10（-8）；显示假检测率的 P 值）与 IBS-D 相关，包括以下内容：神经递质[P2RY4（P=0.001），血管活性肠肽（VIP，P=0.02）]；细胞因子[CCL20（P=0.019）]；免疫功能[C4BPA 补体级联（P=0.0187）]；干扰素相关[IFIT3（P=0.016）]；黏膜修复和细胞黏附[TFF1（P=0.012）]，视黄醇结合蛋白[RBP2（P=0.017）]；纤维连接蛋白（FN1，P=0.009）；和离子通道功能[鸟苷酸环化酶（GUCA2B，P=0.017），PDZ 结构域蛋白 3（PDZD3，P=0.029）]。通过 RT-PCR 验证了 10 个与 IBS-D 病理生理学相关的功能相关的基因。所选基因的折叠变化之间存在显著相关性（Rs=0.73，P=0.013）。P2RY4、GUC2AB、RBP2、FNI 和 C4BPA 基因的上调或下调在 RT-PCR 中得到证实，这也显示出法尼醇 X 受体（FXR）和顶端钠偶联胆汁酸转运体（IBAT/ASBT）的上调。IBS-D 直肠乙状结肠黏膜的 RNA-Seq 和 RT-PCR 分析显示转录组变化，为验证研究提供了依据，以探索黏膜因素在 IBS-D 病理生理学中的作用。
