Stimulus-Secretion Coupling in Diseased Lacrimal Gland

患病泪腺中的刺激-分泌耦合

• 批准号: 6805268
• 负责人: DRISS ZOUKHRI
• 金额: $ 35.66万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6805268
• 关键词: JUN kinase; Sjogren' s syndrome; acetylcholine; biological signal transduction; clinical research; cysteine endopeptidases; disease /disorder model; enzyme inhibitors; enzyme linked immunosorbent assay; female; human subject; immunocytochemistry; interleukin 1; laboratory mouse; lacrimal apparatus; mitogen activated protein kinase; neurotransmitter transport; nitric oxide synthase; secretion

DESCRIPTION (provided by applicant): Sjogren's syndrome, a systemic inflammatory autoimmune disease which occurs almost exclusively in females, is the leading cause of aqueous tear deficient dry eye. To date there is no cure for this disease and the precise mechanisms responsible for the decreased tear secretion are largely unknown. Studies from this laboratory point to a potentially pivotal role of the proinflammatory cytokines, interleukin-1alpha (IL-1a), IL-1beta (IL-lb), and tumor necrosis alpha (TNFa), in the impaired function of the lacrimal gland associated with Sjogren's syndrome. Specifically, we found that these cytokines have a dual target in the lacrimal gland: the nerve endings (i.e., inhibition of neurotransmitter release) and the epithelial cells (i.e., inhibition of lacrimal gland protein secretion). However, the mechanism(s) by which these cytokines interfere with lacrimal gland nerve endings and lacrimal gland acinar epithelial cell functions remain to be elucidated. Thus, the long term objective of the present proposal is to define the intracellular signaling pathways activated by proinflammatory stimuli to inhibit lacrimal gland secretion. A second objective is to test the effects of selective inhibitors of these pathways on lacrimal gland secretion using a murine model of Sjogren's syndrome. To obtain these goals, the following specific aims have been proposed: (1) define the signaling pathways activated by inflammatory stimuli to inhibit neurotransmitter release and lacrimal gland secretion; (2) determine if JNK, ERK, and iNOS are involved in the impaired lacrimal gland secretion associated with Sjogren's syndrome; and (3) determine if ICE activity is necessary for inflammation-induced inhibition of neurotransmitter release and lacrimal gland secretion. Lacrimal gland slices will be prepared from diseased and control female mice. Activation of JNK, ERK, and iNOS will be measured by western blotting. The release of neurotransmitter from lacrimal gland nerve endings and protein secretion will be measured spetrophotometrically. Selective inhibitors of JNK, ERK, and/or iNOS will be given subcutaneously to diseased and control female mice.

描述(申请人提供)：干燥综合征是一种全身性炎症性自身免疫性疾病，几乎只发生在女性，是导致泪水缺乏性干眼的主要原因。到目前为止，这种疾病还没有治愈的方法，导致泪液分泌减少的确切机制在很大程度上还不清楚。该实验室的研究指出，促炎症细胞因子IL-1a(IL-1a)、IL-1b(IL-1b)和肿瘤坏死因子(TNFa)在干燥综合征相关的泪腺功能受损中具有潜在的关键作用。具体地说，我们发现这些细胞因子在泪腺具有双重靶点：神经末梢(即抑制神经递质释放)和上皮细胞(即抑制泪腺蛋白分泌)。然而，这些细胞因子干扰泪腺神经末梢和泪腺腺泡上皮细胞功能的机制(S)仍有待阐明。因此，本提案的长期目标是确定由促炎刺激激活的细胞内信号通路，以抑制泪腺分泌。第二个目标是使用干燥综合征的小鼠模型来测试这些通路的选择性抑制剂对泪腺分泌的影响。为了达到这些目标，已经提出了以下特定的目标：(1)确定炎症刺激激活的信号通路以抑制神经递质释放和泪腺分泌；(2)确定JNK、ERK和iNOS是否参与干燥综合征相关的泪腺分泌受损；以及(3)确定ICE活性是否对于炎症诱导的神经递质释放和泪腺分泌的抑制是必需的。泪腺切片取自患病雌性小鼠和对照雌性小鼠。用免疫印迹法检测JNK、ERK和iNOS的活性。泪腺神经末梢的神经递质释放和蛋白质分泌将用分光光度法测量。JNK、ERK和/或iNOS的选择性抑制剂将被皮下注射给患病和对照雌性小鼠。