Effect of SS-associated cytokines on salivary gland dysfunction

SS相关细胞因子对唾液腺功能障碍的影响

基本信息

批准号：
7788432
负责人：
Olga Juliana Baker
金额：
$ 22.97万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-04 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7788432
关键词：
Acinus organ component Affect Agonist Animal Model Anions Autoimmune Diseases C10 Calcium Carbachol Cell physiology Cells Cholinergic Receptors Chronic Complex Cytosol Data Down-Regulation Dryness Epithelial Epithelial Cells Excision Figs - dietary Functional disorder Generations Gland In Vitro Individual Inflammation Inflammatory Interferons Interleukin-12 Interleukin-18 Interleukin-6 Intestines Ions Lacrimal gland structure Major salivary gland structure Mediating Messenger RNA Minor salivary gland structure Morphology Mus Muscarinics Oral cavity Parotid Gland Patients Permeability Phosphorylation Plasma Production Protein Biosynthesis Proteins Public Health Publishing Rattus Recovery Resistance Role Saliva Salivary Salivary Gland Diseases Salivary Glands Signal Pathway Signal Transduction Sjogren&apos s Syndrome Staging Structure Tight Junctions Tumor Necrosis Factor Activation Tumor Necrosis Factor-alpha Tumor Necrosis Factors Wild Type Mouse airway epithelium apical membrane base claudin-1 protein cytokine in vivo in vivo Model monolayer mouse model nucleotide receptor occludin overexpression protein distribution protein expression public health relevance receptor-mediated signaling saliva secretion

项目摘要

DESCRIPTION (provided by applicant): Sjogren's syndrome (SS) is an autoimmune disorder characterized by inflammation and dysfunction of salivary glands, resulting in impaired secretory function. Levels of the pro-inflammatory cytokines tumor necrosis factor-1 (TNF1), interferon-3 (IFN3), IL-12, interleukin-6 (IL-6), interleukin-18 (IL-18) and interleukin-12 (IL-12) are elevated in salivary glands of patients with SS, although little is known about the effects of these cytokines on salivary epithelial cell tight junction (TJ) integrity which is necessary to establish transepithelial ion gradients that drive saliva secretion. We have demonstrated that chronic exposure of polarized rat parotid gland (Par-C10) epithelial cell monolayers to TNF1 and/or IFN3 decreases transepithelial resistance (TER) and transepithelial anion secretion induced by carbachol, a muscarinic cholinergic receptor agonist, or UTP, a P2Y2 nucleotide receptor agonist. In contrast, TNF1 and/or IFN3 had no effect on agonist-induced increases in the intracellular calcium concentration [Ca2+]i in Par-C10 cells indicating that individual cell signaling is unaffected by cytokines. Our studies show that among the TJs, claudin-1 is selectively downregulated by TNF1 and/or IFN3. In cells treated with TNF1, claudin-1 downregulation returns to normal levels upon incubation of cells for 24 h in cytokine- free medium. Under these conditions, recovery of claudin-1 expression corresponds with increases in TER and agonist-induced short circuit current (Isc). We have obtained new preliminary data demonstrating that silencing of claudin-1 expression decreases TER in Par-C10 monolayers. Furthermore, the cellular distribution of the TJ proteins occludin and ZO-1 was altered by TNF1 and/or IFN3 treatment of Par- C10 three-dimensional (3D) acinar spheres. Based on these preliminary findings, we will examine the overall hypothesis that cytokine- induced changes in the expression and/or distribution of TJ proteins affects TJ integrity in Par-C10 cell monolayers, consistent with a loss in saliva secretion associated with cytokine generation in the intact parotid gland. In addition, we propose to identify the cellular mechanisms underlying cytokine-induced disruption of TJ integrity using in vitro and in vivo models of SS. Studies in Specific Aim 1 will characterize the mechanisms underlying cytokine-induced decreases in claudin-1 expression. Studies in Specific Aim 2 will identify the TNF1- and IFN3-mediated changes in TJ protein phosphorylation and removal from the TJ complex in Par-C10 cells. Studies in Specific Aim 3 will characterize TJ mRNA and protein expression, TJ morphology, and TJ cellular distribution in parotid glands of the C57BL/6.NOD-Aec1Aec2 mouse model of SS and mice overexpressing TNF1, as compared to wild type mice. These studies will provide a greater understanding the mechanisms whereby saliva secretion is decreased during the pro-inflammatory stages of salivary gland diseases, a significant initial step in defining the poorly-studied signaling pathways that regulate TJ structural integrity in parotid acini. PUBLIC HEALTH RELEVANCE: Sjogren's syndrome (SS) is an autoimmune disease characterized by salivary gland dysfunction leading to severe dryness of the oral cavity. Pro-inflammatory cytokines are up-regulated in plasma and in salivary glands from patients with SS, however, little is known of their role in salivary gland dysfunction. We propose to determine the role of pro-inflammatory cytokines in salivary gland tight junction integrity causing reduction of saliva secretion.

描述（由申请人提供）：Sjogren综合征（SS）是一种自身免疫性疾病，其特征是唾液腺发炎和功能障碍，导致分泌功能受损。促炎性细胞因子的肿瘤坏死因子-1（TNF1），干扰素-3（IFN3），IL-12，IL-12，肠介菌-6（IL-6），IL-6（IL-6），肠间介体-18（IL-18）（IL-18）和白介素-12（IL-12）几乎没有众所周知的唾液膜，这些cy在S唾液中，这些cy均未受到s囊的影响。细胞紧密连接（TJ）完整性，这是建立驱动唾液分泌的跨越离子梯度所必需的。我们已经证明，慢性暴露于TNF1和/或IFN3上的极化大鼠上皮细胞单层降低了transepithelial耐药性（TER）和transepithepithial阴离子的分泌，由卡尔巴乔（Carbachol）诱导的carbachol，carbachol，carbachol，肌液胆碱性受体受体agonist agonisist agonisist og agonisist和utp p2y2。相反，TNF1和/或IFN3对PAR-C10细胞中细胞内钙浓度[Ca2+] I的增加没有影响，这表明单个细胞信号传导不受细胞因子的影响。我们的研究表明，在TJ中，Claudin-1被TNF1和/或IFN3选择性下调。在用TNF1处理的细胞中，Claudin-1下调在无细胞因子 - 无细胞因子培养基中孵育24小时后，恢复到正常水平。在这些条件下，Claudin-1表达的恢复与TER和激动剂诱导的短路电流（ISC）的增加相对应。我们获得了新的初步数据，表明Claudin-1表达的沉默会降低PAR-C10单层中的TER。此外，通过TNF1和/或IFN3处理三维（3D）腺泡球的TNF1和/或IFN3处理，TJ蛋白的细胞分布改变了。基于这些初步发现，我们将研究总体假设，即细胞因子诱导的TJ蛋白的表达和/或分布的变化会影响PAR-C10细胞单层中的TJ完整性，这与整体羊皮腺中与细胞因子产生相关的唾液分泌损失一致。此外，我们建议使用SS的体外和体内模型来确定细胞因子诱导的TJ完整性破坏的细胞机制。特定目标1中的研究将表征细胞因子诱导的Claudin-1表达降低的机制。特定目标2中的研究将鉴定TJ蛋白磷酸化的TNF1-和IFN3介导的变化，并从PAR-C10细胞中的TJ复合物中去除。与野生型小鼠相比，在特定目标3中的研究将表征TJ mRNA和蛋白质表达，TJ形态和TJ细胞分布。SS和过表达TNF1的小鼠的NOD-AEC1AEC2小鼠模型，与野生型小鼠相比。这些研究将提供更大的理解，即在唾液腺疾病的促炎阶段唾液分泌减少的机制，这是确定调节羊毛细胞acini中TJ结构完整性的不良信号通路的重要第一步。公共卫生相关性：Sjogren综合征（SS）是一种自身免疫性疾病，其特征是唾液腺功能障碍，导致口腔严重干燥。促炎性细胞因子在血浆和SS患者的唾液腺中被上调，但是，对它们在唾液腺功能障碍中的作用知之甚少。我们建议确定促炎细胞因子在唾液腺紧密连接完整性中的作用，从而减少唾液分泌。