Conjunctival Goblet Cell Mucin Secretion in Inflammation and Its Resolution

项目摘要/摘要 作为对环境的反应，结膜杯状细胞的功能是维持健康的泪膜和眼睛。 表面，预防炎症性疾病。杯状细胞分泌粘蛋白MUC5AC，这种粘蛋白对 眼表。杯状细胞通过积极地保持泪膜的粘液层来维持泪膜的最佳粘液层。 处于正常的健康水平。然而，在疾病中，粘蛋白可能会过度产生，就像过敏性结膜炎一样。我们的 实验室专注于杯状细胞粘蛋白产生在健康和炎症性疾病中的作用 过敏性结膜炎。特殊的前解析分子(SPM)由omega-3脂肪酸产生 例如DHA。有一种复杂且受到严格监管的生物学方法，即使用支持拆分的分子来控制 杯状细胞分泌粘蛋白，维持眼表动态平衡。在疾病中，有一种由过敏介导的 粘蛋白的失调/过度生产，有助于SPM解决的病理生理学问题。我们的整体 目的是研究SPM是如何由结膜产生的，并以性别依赖的方式在 结膜杯状细胞中调节健康和疾病中粘蛋白分泌的分子水平。我们将重点关注 泪液和结膜中存在的解析素D Rvd1-6。RvD1和RvD2刺激杯状细胞 功能。在具体目标1中，我们将重点关注健康，并确定哪些RvD家庭成员(RvD3-6) 结膜细胞内[Ca~(2+)]_i升高，cAMP升高，粘蛋白分泌增加 杯状细胞。对于这三个目标，我们将评估是否存在性别相关的反应差异。在……里面 具体目标2我们将研究在健康状态下Rvd1是否使用带有GPR32(受体)/DHA的自分泌回路 (前体)/RvD1轴控制其生物合成。在特定目标3中，我们将研究疾病和过敏 结膜炎询问RvD家族成员(RvD2-6)哪些抗炎症介质刺激 杯状细胞增加细胞内[Ca~(2+)]i和分泌量，促进体内AED的分解。我们将使用人类 培养的结膜杯状细胞，两种类型的SPM受体敲除小鼠，以及一种过敏性小鼠模型 眼疾。细胞内[Ca~(2+)]、cAMP、MUC5AC分泌、DHA释放和RvD1生物合成将 量过了。在动物模型中，临床症状、MUC5AC的分泌、白细胞的转运都会 下定决心。

Project Summary/Abstract In response to the environment, the conjunctival goblet cells function to maintain a healthy tear film and ocular surface, and to prevent inflammatory diseases. Goblet cells secrete the mucin MUC5AC that is protective to the ocular surface. Goblet cells maintain an optimal mucous layer of the tear film by actively keeping it at normal levels in health. In disease, however, mucins can be over produced as in allergic conjunctivitis. Our laboratory concentrates on the role of goblet cell mucin production in health and in the inflammatory disease allergic conjunctivitis. The specialized pro-resolving molecules (SPMs) are produced from omega-3 fatty acids such as DHA. There is a complex and tightly regulated biology of using pro-resolution molecules to control goblet cell mucin secretion and maintain ocular surface homeostasis. In disease there is an allergy-mediated dysregulation/overproduction of mucins that contribute to the pathophysiology that SPMs resolve. Our overall goal is to investigate how SPMs are produced by the conjunctiva and act in a sex-dependent manner at a molecular level in conjunctival goblet cells to regulate mucin secretion in health and disease. We will focus on the resolvin Ds RvD1-6 that are present in tears and conjunctiva. RvD1 and RvD2 stimulate goblet cell function. In Specific Aim 1 we will focus on health and determine which RvD family members (RvD3-6) increase the intracellular [Ca2+] ([Ca2+]i), elevate cAMP, and stimulate mucin secretion in cultured conjunctival goblet cells. For this and all three aims we will evaluate if there a sex-dependent difference in response. In Specific Aim 2 we will investigate if in health RvD1 uses an autocrine circuit with a GPR32 (receptor) /DHA (precursor)/RvD1 axis to control its biosynthesis. In Specific Aim 3 we will study disease and in allergic conjunctivitis interrogate which RvD family members (RvD2-6) counter pro-inflammatory mediator stimulated goblet cell increase in [Ca2+]i and secretion, and promote resolution of AED in vivo. We will use human cultured conjunctival goblet cells, two types of SPM receptor knock out mice, and a mouse model of allergic eye disease. Intracellular [Ca2+], cAMP, MUC5AC secretion, DHA release, and RvD1 biosynthesis will be measured. In the animal model clinical symptoms, MUC5AC secretion, and leukocyte trafficking will be determined.