Inflammatory Bowel Disease: A Novel Therapeutic Approach

描述（由申请人提供）：溃疡性结肠炎（UC）和克罗恩病（CD）是胃肠道慢性，复发性炎症性疾病，统称为炎症性肠病（IBD）。在美国，估计有100万人患有IBD，其中包括儿童和青少年，病因在很大程度上仍然未知。IBD的发病机制通常被认为是对普遍存在的非致病性微生物抗原的异常免疫反应的影响。鉴于IBD的传统和新型治疗方法，它们的副作用和高成本促使人们研究新的替代治疗方式。其中一种替代品是姜黄香料中一种众所周知的抗炎和化学预防成分，姜黄素，它最近才被报道能显著减轻或改善实验诱导的小鼠结肠炎。姜黄素吸收不良，在胃肠道中代谢广泛，这可能是报道中缺乏毒性的原因。本项目的目的是进一步研究姜黄素对结肠炎的保护作用机制，并为儿童和成人IBD患者的临床试验提供充分的科学背景。这将通过研究膳食姜黄素在免疫型结肠炎小鼠模型中的有效性来完成。将使用具有不同免疫失调倾向的IL-10-/-和tcrα -/-小鼠（分别以Th1-和th2为主）。为了进一步了解姜黄素的分子靶点及其作用机制，将在宏观、形态学和生化评价之后，对结肠上皮细胞和免疫细胞的基因表达谱进行比较全基因组微阵列分析。此外，在HPLC/LC-MS技术的帮助下，将对从对照小鼠和已建立差异极化炎症的小鼠分离的结肠炎细胞中姜黄素的上皮代谢进行定性和定量分析。研究人员将利用小鼠巨噬细胞（RAW264.7）和结肠炎细胞（YAMC细胞）模型评估姜黄素新代谢产物的抗炎活性。为了进一步揭示姜黄素在肠道中作用的新机制，我们将研究其对结肠抗菌隐素相关肽表达的影响。此外，我们将描述其作用于已确定的独特调控基因的机制，如lps介导的MIP-2（小鼠IL-8同源物）基因转录的早期诱导。总之，该项目将为姜黄素在IBD中的作用机制、其在不同极化t细胞介导炎症中的分子靶点以及健康和炎症结肠中活性化合物的分子形式提供新的信息。

DESCRIPTION (provided by applicant): Ulcerative Colitis (UC) and Crohn's Disease (CD) are chronic, relapsing inflammatory disorders of the gastrointestinal tract, collectively known as Inflammatory Bowel Disease (IBD). The cause of IBD, which in U.S. affects an estimated 1 million individuals, including children and adolescents, remains largely unknown. Pathogenesis of IBD is most commonly viewed as an effect of an aberrant immune response against ubiquitous, non-pathogenic microbial antigens. In view of traditional and novel treatments of IBD, their side effects and high cost prompt investigations into new, alternative treatment modalities. One such alternative is a long-known anti-inflammatory and chemopreventive component of turmeric spice, curcumin, which was only recently reported to significantly attenuate or ameliorate experimentally induced colitis in mice. Curcumin is poorly absorbed, and extensively metabolized in the gastrointestinal tract, which may account for the reported lack of toxicity. The goal of this proposed project is to investigate the mechanism of curcumin's protective action against colitis in further detail and to provide a sufficient scientific background for clinical trials in both pediatric and adult IBD patients. This will be accomplished by studying the effectiveness of dietary curcumin in immune-based mouse models of colitis. IL-10-/- and TCRalpha-/- mice with different propensities for immune dysregulation (Th1- and Th2-dominated, respectively) will be used. Macroscopic, morphological, and biochemical evaluation will be followed by comparative genome-wide microarray analyses of gene expression profiles in colonic epithelial and immune cells in order to gain further insight into molecular targets of curcumin and its mechanism of action. Also, epithelial metabolism of curcumin will be qualitatively and quantitatively analyzed in colonocytes isolated from control mice and from mice with established differentially polarized inflammation with the help of HPLC/LC-MS technology. The compounds identified as novel metabolites of curcumin will be evaluated for anti-inflammatory activity utilizing established models of mouse macrophages (RAW264.7) and colonocytes (YAMC cells). To shed more light into novel mechanisms of curcumin action in the intestine, its effects on expression of colonic antibacterial cryptdin -related peptides will be investigated. Also, we will describe its mechanism of action on identified uniquely regulated genes such as LPS-mediated early induction of MIP-2 (murine homologue of IL-8) gene transcription. In summary, the proposed project will provide novel information about the mechanism of curcumin's action in IBD, about its molecular targets in differently polarized T-cell mediated inflammation as well as about the molecular forms of the active compound in the healthy and inflamed colon.