Sex difference in conjunctival epithelial cell response and their profile of extracellular bioactive lipids duringallergic inflammation

项目总结 细胞外生物活性脂质介体的性别差异，尤其是专门化的 炎症过程中的促分解和促炎脂质介质(分别为SPM和PIM) 与性别显著相关的细胞反应差异，以及各种疾病的患病率和严重程度 炎症性疾病，包括眼表炎症。 结膜上皮作为眼表组织，排列在眼睑内侧，覆盖巩膜， 从而为眼睛的前段提供稳定性。结膜杯状细胞分泌粘蛋白。 湿润和保护因炎症引起的分泌不足或过多而导致视力的眼表 骚乱。因此，结膜炎症的时间依赖性消退对透明角膜至关重要。 和清晰的视野。 我们令人兴奋的初步数据显示，CGCs以细胞外小泡(EV)的形式分泌SPM和PIM 为了维持体内平衡，在组胺介导的过敏过程中，SPM的数量急剧增加 回应。有趣的是，这种增加只在女性CGC的EV中观察到，而不是男性。收集到的电动汽车 在相同条件下，雄性的SPM数量与对照组相比略有下降。 为了进一步研究这些新发现，这项研究计划旨在研究儿童的性别差异 人脐静脉内皮细胞的细胞反应及其通过EVS分泌的生物活性脂质介质 组胺介导的过敏性炎症。对于目标1，我们将通过以下方式识别细胞反应中的性别差异 定量粘蛋白分泌量和细胞内钙浓度的变化，并测量 参与过敏反应和生物活性脂质介体合成的细胞内酶的激活。 对于目标2，我们将确定在细胞外生物活性脂质介质的分布中的性别差异 通过分析电动汽车中装载的SPM和PIM的类型和数量以及它们的时间进程来实现过敏反应 分泌物。 这项研究的成功将为生物活性脂质介导的眼表提供新的见解 结膜产生的保护机制和基础为量身定做的性别依赖，以脂质为基础 视力衰弱眼表疾病的治疗方法。

PROJECT SUMMARY Sex difference in the profile of extracellular bioactive lipid mediators, especially that of the specialized pro-resolving and proinflammatory lipid mediators (SPMs and PIMs, respectively), during inflammation is significantly correlated to sex differences in the cellular response, and prevalence and the severity of various inflammatory diseases, including ocular surface inflammation. As an ocular surface tissue, conjunctival epithelium lines the inside of the eyelids and covers the sclera, thus providing stability to the anterior segment of the eye. The conjunctival goblet cells (CGCs) secrete mucins to wet and protect the ocular surface, of which under- or over-secretion due to inflammation causes visual disturbance. Therefore, time-dependent resolution of conjunctival inflammation is critical for a transparent cornea and clear vision. Our exciting preliminary data show that CGCs secrete SPMs and PIMs as extracellular vesicle (EV) cargo to maintain homeostasis, and the amount of SPMs dramatically increases during a histamine-mediated allergic response. Interestingly, the increase was only observed in EVs of female CGCs but not males. EVs collected from males under the same condition showed a slight decrease in the amounts of SPMs compared to the control. To further investigate these novel findings, this research proposal aims to study the sex differences in the cellular response of human CGCs and the bioactive lipid mediators that CGCs secrete via EVs during histamine-mediated allergic inflammation. For Aim 1, we will identify sex differences in the cellular response by quantifying the magnitude of mucin secretion and change in intracellular Ca2+ concentration, and measuring the activation of intracellular enzymes involved in the allergic response and synthesis of bioactive lipid mediators. For Aim 2, we will determine the sex differences in the profile of extracellular bioactive lipid mediators during the allergic response by analyzing the type and amount of SPMs and PIMs loaded into EVs, and their time course of secretion. The success of this study will provide new insights into the bioactive lipid-mediated ocular surface protective mechanism generated by the conjunctiva and the basis for a tailored sex-dependent, lipid-based therapeutic approach in the management of vision-debilitating ocular surface diseases.