Restraint stress-induced neurogenic bladder inflammation

约束应激诱发的神经源性膀胱炎症

• 批准号: 7060529
• 负责人: THEOHARIS C. THEOHARIDES
• 金额: $ 23.99万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7060529
• 关键词: autoradiography; corticotropin releasing factor; female; gel mobility shift assay; genetically modified animals; histamine; hormone regulation /control mechanism; inflammation; interleukin 6; laboratory mouse; light microscopy; mast cell; neurogenic urinary bladder disorder; neurotensin; nuclear factor kappa beta; stress; substance P; tumor necrosis factor alpha; vascular endothelium permeability

DESCRIPTION (provided by applicant): The urinary bladder is often the site of subacute or chronic inflammation, in the absence of infection, as in interstitial cystitis (IC), a painful bladder disorder occurring mostly in women. Symptoms of urinary frequency and pelvic pain commonly worsen perimenstrually and under stress in IC. Bladder mastocytosis with mast cell activation has been documented in IC. We also showed that acute immobilizationstress in rodents induced bladder mast cell activation, a process that was dependent on the neuropeptides neurotensin (NT) and substance P (SP), as it was absent in rodents treated with capsaicin to deplete sensory nerve fibers of their SP content and was also inhibited by the NT receptor antagonist SR48692. Moreover, pretreatment of bladder with estradiol increased the stimulatory effect of SP, by activating high affinity estrogen receptors that we have identified on bladder mast cells. It was recently shown that bladder inflammation could not occur in mast cell deficient mice infected with the neurotropic pseudorabies virus. Mast cells are located perivascularly close to nerve processes and may secrete many vasoactive, proinflammatory and neurosensitizing molecules in response to allergic triggers, as well as by direct nerve stimulation and by acute immobilization stress. Corticotropin releasing hormone (CRH) is released from the hypothalamus under stress and activates the hypothalamic-pituitary-adrenal (HPA) axis. However, both CRH and its structurally related urocortin (Ucn) are also released in the periphery where they have proinflammatory effects. CRH and Ucn induced rat skin mast cell activation and increased vascular permeability, both of which were inhibited by pretreatment with neutralizing antiserum to CRH or the CRH-receptor (CRH-R) antagonist, antalarmin. CRH or acute stress-induced skin vascular permeability was absent in W/W v mast cell deficient mice, but was present in their +/+ controls indicating it is mast cell dependent. Acute stress also triggered rat bladder mast cell activation that was blocked by a NT-receptor antagonist. The proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosisfactor-alpha (TNF-alpha) were recently shown to be elevated in urine of IC patients. We are hypothesizing that acute stress releases (CRH) and/or (Ucn) in the bladder leading, directly or through SP or NT, to mast cell activation, increased vascular permeability and the expression of proinflammatory molecules. We propose to use normal and genetically deficient female mice to investigate the effect of acute stress and CRH/Ucn on: (1) bladder mast cell and urothelial Nuclear Factor kappa B (NF-kappaB)activation, as well as the levels of histamine,lL-6 and TNF-alpha in the urine collected from an indwelling catheter; (2) bladder vascular permeability quantitated by 99Technetium-gluceptate (99Tc) extravasation; (3) Vascular permeability, urine mediator release, as well as NF-kappaB activation in W/W v mast cell deficient mice, as well as in CRH knock-out mice and their +/+ controls; (4) mouse bladder mast cell and urothelial NF-KB activation, as well as secretion of histamine, IL-6 or TNF-alpha induced by intravesical administration of CRH/Ucn. These studies will help us understand how acute stress triggers bladder mast cell activation leading to increased vascular permeability and proinflammatory molecule release. Our findings may be relevant to the pathophysiology of IC and may suggest new therapeutic approaches.

描述（由申请人提供）：在无感染的情况下，膀胱通常是亚急性或慢性炎症的部位，如间质性膀胱炎（IC），一种主要发生于女性的疼痛性膀胱疾病。IC患者的尿频和盆腔痛症状通常在月经期和应激状态下加重。膀胱肥大细胞增多症伴肥大细胞活化已在IC中记录。我们还表明，急性immobilizationstress在啮齿类动物诱导膀胱肥大细胞活化，这是一个依赖于神经肽神经降压素（NT）和P物质（SP）的过程，因为它是缺乏在啮齿类动物与辣椒素处理，以耗尽其SP含量的感觉神经纤维，也被抑制NT受体拮抗剂SR 48692。此外，用雌二醇预处理膀胱增加了SP的刺激作用，通过激活我们已经确定的膀胱肥大细胞上的高亲和力雌激素受体。最近的研究表明，在嗜神经性伪狂犬病病毒感染的肥大细胞缺陷小鼠中不会发生膀胱炎症。肥大细胞位于血管周围接近神经过程，并可分泌许多血管活性，促炎和神经增敏分子，以响应过敏触发，以及通过直接神经刺激和急性制动应激。促肾上腺皮质激素释放激素（CRH）在应激状态下从下丘脑释放并激活下丘脑-垂体-肾上腺（HPA）轴。然而，CRH及其结构相关的尿皮质素（Ucn）也在外周释放，在外周它们具有促炎作用。CRH和UCN诱导大鼠皮肤肥大细胞活化和血管通透性增加，这两个抑制预处理与中和抗血清CRH或CRH受体（CRH-R）拮抗剂，antalarmin。CRH或急性应激诱导的皮肤血管通透性在W/W v肥大细胞缺陷小鼠中不存在，但在其+/+对照中存在，表明其是肥大细胞依赖性的。急性应激也引发大鼠膀胱肥大细胞活化，这被NT受体拮抗剂阻断。最近发现IC患者尿中的促炎细胞因子白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）升高。我们假设膀胱中的急性应激释放（CRH）和/或（Ucn）直接或通过SP或NT导致肥大细胞活化、血管通透性增加和促炎分子的表达。本研究拟利用正常和遗传缺陷雌性小鼠，研究急性应激和CRH/Ucn对以下指标的影响：（1）膀胱肥大细胞和尿路上皮细胞核因子κ B（2）通过99锝-葡庚糖酸盐（99 Tc）外渗定量的膀胱血管渗透性;（3）W/Wv肥大细胞缺陷小鼠以及CRH敲除小鼠及其+/+对照中的血管通透性、尿介质释放以及NF-κ B活化;（4）膀胱内施用CRH/Ucn诱导的小鼠膀胱肥大细胞和尿路上皮NF-κ B活化以及组胺、IL-6或TNF-α分泌。这些研究将帮助我们了解急性应激如何触发膀胱肥大细胞激活，导致血管通透性增加和促炎分子释放。我们的研究结果可能与IC的病理生理学有关，并可能提出新的治疗方法。