Role of MCH in Adipose Tissue & Intestinal Inflammation

描述（由申请人提供）：黑色素浓集激素（MCH）是一种调节食欲和体重的下丘脑神经肽，也已被证明在各种病因的结肠炎中发挥促炎作用。因此，肠系膜人前脂肪细胞表达MCH受体，MCH与这些受体的结合刺激IL-8的表达，IL-8是一种参与IBD发病机制的中性粒细胞化学引诱物。由于克罗恩病患者的肠肠系膜脂肪包裹与促炎细胞因子表达增加相关，因此MCH可能参与了这种炎症反应。本研究的目标有两个：1）检查MCH将结肠炎期间粘膜炎症与肠系膜脂肪库炎症变化联系起来的假设，以及2）NF-kB和MAP激酶途径有助于相关信号传导。这将通过比较小鼠脂肪库中蛋白质和mRNA水平的MCH和MCH受体表达与TNBS-、DSS-和IL-10 KO结肠炎期间的粘膜炎症程度，通过激光捕获显微切割、免疫组织化学分析和共聚焦显微镜鉴定该组织中表达MCH/MCHR 1的细胞，并通过蛋白质印迹、RT-PCR和ELISA解剖信号传导途径来完成。

DESCRIPTION (provided by applicant): Melanin-concentrating hormone (MCH), a hypothalamic neuropeptide that regulates appetite and body weight, has also been shown to play a proinflammatory role in colitis of various etiologies. Thus, mesenteric human preadipocytes express MCH receptors, and MCH binding to these receptors stimulates expression of IL-8, a neutrophil chemoattractant involved in IBD pathogenesis. Since patients with Crohn's disease have mesenteric fat wrapping of the bowel associated with increased expression of proinflammatory cytokines, it is likely that MCH participates in this inflammatory response. The goals of this study are two-fold: 1) To examine the hypothesis that MCH links mucosal inflammation with inflammatory changes in the mesenteric fat depots during colitis and 2) that NF-kB and MAP kinase pathways contribute to the signaling involved. This will be done by comparing MCH and MCH-receptor expression at the protein and mRNA levels in murine fat depots to degree of mucosal inflammation during TNBS-, DSS-, and IL-10 KO colitis, identifying the cells expressing MCH/MCHR1 in this tissue through laser capture microdissection, immunohistochemical analysis, and confocal microscopy and dissecting signalling pathways by western blot, RT-PCR and ELISA.