Mast cells, antidepressants and chronic fatigue syndrome

描述（由申请人提供）：慢性疲劳综合征（CFS）的特征是疲劳、不适、睡眠和自主神经紊乱;它被认为是一种神经免疫疾病，伴有下丘脑-垂体-肾上腺（HPA）轴失调，由压力引起，并与高度残疾相关。CFS通常与纤维肌痛、肠易激综合征（IBS）、间质性膀胱炎（IC）和偏头痛等共病疾病一起发生，所有这些疾病也会因压力而恶化。CFS没有可靠的动物模型。肥大细胞已成为受应激影响的神经免疫内分泌过程的主要调节因子，并与CFS相关的所有共病疾病有关。我们已经证明：（a）肥大细胞与神经末梢具有功能关联;（B）急性应激激活肥大细胞，该作用被促肾上腺皮质激素释放激素（CRH）中和抗血清预处理阻断，（c）应激增加血脑屏障（BB B）通透性，其被CRH-受体-1（CRH-R1）拮抗剂Antalarmin抑制，并且在肥大细胞缺陷的W/W小鼠中不发展，（d）人肥大细胞表达CRH受体，CRH受体的激活导致血管内皮生长因子（VEGF）的选择性释放。（e）CRH对肥大细胞的一些刺激作用由神经降压素（NT）介导，神经降压素（NT）已显示调节HPA轴。三环类抗抑郁药对CFS和其他共病有帮助，但其作用机制尚不清楚。我们的初步结果表明，三环类抗抑郁药阿米替林可以抑制大鼠肥大细胞分泌和细胞内钙离子水平。假设：CRH，或结构相关的尿皮质素（Ucn），分泌的压力激活间脑肥大细胞，无论是单独或与其他神经肽，如NT，导致释放的分子，有助于中央发病机制的CFS，分泌可以抑制三环类抗抑郁药。我们将研究：目标1。剂量-反应（0.1-100 uM）和时间进程（0.5，1，6，24 h）三种不同类型的抗抑郁药的作用，（a）三环类抗抑郁药（阿米替林，丙咪嗪），（B）选择性5-羟色胺摄取抑制剂（c）安非他酮对组胺、IL-1、IL-6、IL-8、IL-13、TNF、类胰蛋白酶和VEGF从正常人脐带来源的培养肥大细胞（hCBMCs）衍生自CD 34+祖细胞触发IL-1，CRH或Ucn（100 nM）。目标二。这些抗抑郁药的作用显示在Aim 1中有效，因为它们抑制“脑肥大细胞”的能力，所述“脑肥大细胞”是通过在10 nM IL-4和神经生长因子（NGF）存在下培养人脐带基质干细胞（hCMSC）而产生的，所述人脐带基质干细胞是CD 34-，如在Aim 1 +/- NT（0.1-100 mM）中那样刺激。这些研究的结果将进一步加深我们对应激激素释放的分子的理解，以及哪些抗抑郁药可能有助于抑制这些作用。未来的研究将建立在这些发现的基础上，开发CFS的体外和体内模型，并导致选择抗抑郁药或其他抑制脑肥大细胞的分子进行临床试验。

DESCRIPTION (provided by applicant): Chronic fatigue syndrome (CFS) is characterized by fatigue, malaise, sleep and autonomic disturbances; it is considered a neuroimmune disorder with dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis, precipitated by stress and associated with high disability. CFS often occurs with comorbid diseases such as fibromyalgia, irritable bowel syndrome (IBS), interstitial cystitis (IC) and migraines, all of which also worsen by stress. There are no reliable animal models for CFS. Mast cells have emerged as a major regulator of neuroimmune endocrine processes affected by stress and have been implicated in all comorbid diseases associated with CFS. We have shown that: (a) mast cells have functional associations with nerve endings; (b) acute stress activates mast cells, an action blocked by pretreatment with corticotropin-releasing hormone (CRH) neutralizing antiserum, (c) stress increases blood-brain-barrier (BBB) permeability, which is inhibited by the CRH-receptor-1 (CRH-R1) antagonist Antalarmin and does not develop in mast cell deficient W/W mice, (d) human mast cells express CRH receptors, activation of which leads to selective release of vascular endothelial growth factor (VEGF), (e) some of the stimulatory effects of CRH on mast cells are mediated by neurotensin (NT), which has been shown to regulate the HPA axis. Tricyclic antidepressants are helpful in CFS and in the other comorbid diseases, but this mechanism of action is unknown. Our preliminary results show that the tricyclic antidepressant amitriptyline can inhibit rat mast cell secretion and intracellular calcium ion levels. Hypothesis: CRH, or the structurally related urocortin (Ucn), secreted by stress activates diencephalic mast cells, either alone or together with other neuropeptides such as NT leading to release of molecules that contribute to the central pathogenesis of CFS, and secretion of which can be inhibited by tricyclic antidepressants. We will investigate: Aim 1. The dose-response (0.1-100 uM) and time-course (0.5,1,6, 24 h) effects of three different classes of antidepressants, (a) the tricyclic (amitriptyline, imipramine), (b) the selective serotonin uptake inhibitors (fluoxetine, sertraline) and (c) bupropion on secretion of histamine, IL-1, IL-6, IL-8, IL-13, TNF, tryptase and VEGF from normal human umbilical cord-derived cultured mast cells (hCBMCs) derived from CD34+ progenitors triggered by IL-1, CRH or Ucn (100 nM). Aim 2. The effect of those antidepressants shown to be effective in Aim 1 for their ability to inhibit "brain mast cells" developed by culturing human umbilical cord matrix stem cells (hCMSCs) that are CD34- in the presence of 10 nM IL-4 and nerve growth factor (NGF), stimulated as in Aim 1 +/- NT (0.1-100 mM). Results from these studies will further our understanding of molecules released in response to stress hormones and which antidepressants may be useful in inhibiting these effects. Future studies will build on these findings to develop in vitro and in vivo models of CFS and lead to clinical trials with select antidepressants or other molecules that inhibit brain mast cells.