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）是胃肠道的慢性复发性炎症性疾病，统称为炎症性肠病（IBD）。IBD的病因在美国影响着估计100万人，包括儿童和青少年，但在很大程度上仍然未知。IBD的发病机制最常被视为针对普遍存在的非致病性微生物抗原的异常免疫应答的影响。鉴于IBD的传统和新型治疗方法，其副作用和高成本促使人们研究新的替代治疗方式。一种这样的替代品是姜黄香料的长期已知的抗炎和化学预防成分姜黄素，其仅在最近被报道显著减弱或改善小鼠中实验诱导的结肠炎。姜黄素吸收不良，在胃肠道中广泛代谢，这可能是报告缺乏毒性的原因。该项目的目标是进一步详细研究姜黄素对结肠炎的保护作用机制，并为儿科和成人IBD患者的临床试验提供足够的科学背景。这将通过研究饮食姜黄素在基于免疫的结肠炎小鼠模型中的有效性来实现。将使用具有不同免疫失调倾向（分别为Th 1和Th 2主导）的IL-10-/-和TCR α-/-小鼠。宏观，形态学和生化评价之后，将比较全基因组微阵列分析的基因表达谱在结肠上皮细胞和免疫细胞，以获得进一步了解姜黄素的分子靶点及其作用机制。此外，将在HPLC/LC-MS技术的帮助下，定性和定量分析从对照小鼠和具有建立的差异极化炎症的小鼠分离的结肠细胞中姜黄素的上皮代谢。将利用已建立的小鼠巨噬细胞（RAW264.7）和结肠细胞（YAMC细胞）模型评价被鉴定为姜黄素新代谢物的化合物的抗炎活性。为了更清楚地揭示姜黄素在肠中作用的新机制，将研究其对结肠抗菌穴蛋白相关肽表达的影响。此外，我们将描述其作用机制，确定独特的调控基因，如LPS介导的MIP-2（小鼠同源的IL-8）基因转录的早期诱导。总之，拟议的项目将提供有关姜黄素在IBD中的作用机制的新信息，有关其在不同极化T细胞介导的炎症中的分子靶点，以及有关健康和发炎结肠中活性化合物的分子形式。

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.