IL-19促进Eos趋化因子表达在CRSwNP发病中的作用机制研究
批准号：
82101194
项目类别：
青年科学基金项目（C类）
资助金额：
30.0 万元
负责人：
黄子真
依托单位：
学科分类：
嗅觉、鼻及前颅底疾病
结题年份：
2024
批准年份：
2021
项目状态：
已结题
项目参与者：
黄子真
项目摘要
结项摘要
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中文摘要
嗜酸性粒细胞（Eos）浸润是慢性鼻窦炎伴鼻息肉（CRSwNP）的难治因素之一，发生机制未明。我们发现CRSwNP鼻黏膜中Eos浸润程度高，多种Eos趋化因子、IL-19及其特异性受体高表达，IL-19与ECP表达正相关；IL-19促进源于人鼻黏膜的原代上皮细胞和成纤维细胞表达RANTES，促进上皮细胞表达LTB4。推测IL-19上调Eos趋化因子表达参与CRSwNP Eos浸润。本研究拟以IL-19刺激原代细胞及人鼻黏膜组织块，测Eos趋化因子、相关信号通路蛋白及Eos浸润数量；IL-19刺激/封闭联合信号通路抑制/活化刺激原代细胞，测Eos趋化因子；封闭IL-19或其受体，评估Eos浸润型慢性鼻窦炎小鼠的鼻窦炎症状、鼻黏膜上皮Eos浸润数量、Eos趋化因子及通路蛋白。从人体组织标本、体内外实验分别证明IL-19上调Eos趋化因子促进Eos浸润，进一步完善CRSwNP中Eos浸润的发病机制
英文摘要
Eosinophil (Eos) infiltration is one of the important refractory factors of chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP). We found that CRSwNP had a high degree of Eos infiltration in nasal mucosa, and several Eos chemokines including Eotaxin, LTB4 and RANTES, and IL-19 and its specific receptors IL-20R1/IL-20R2 were highly expressed in nasal mucosa, too. IL-19 was positively correlated with ECP expression. IL-19 promoted RANTES expression in primary epithelial cells and fibroblasts derived from human nasal mucosa, and also promoted LTB4 expression in epithelial cells. It is speculated that IL-19 promotes the expression of Eos chemokines and participates in the Eos infiltration of CRSwNP. The aim of this study was to measure the expression of Eos chemokines and the activation of related signal pathways in primary epithelial cells and fibroblasts derived from human nasal mucosa stimulated with IL-19, and to calculate the number of Eos infiltration in human nasal mucosa tissue injected with IL-19 and co-cultured with peripheral blood leukocytes; and to measure the expression of Eos chemokines in primary epithelial cells and fibroblasts stimulated with IL-19 or anti-IL-19 following stimulated with inhibition or activation of the related signal pathways; and to assess the CRS symptoms and the number of Eos infiltration in nasal mucosa and the Eos chemokines and signal pathways after blocking IL-19 or its specific receptor IL-20R1 in mice with eosinophilic CRS. This study aims to prove that IL-19 up-regulates the Eos chemokines and promotes the Eos infiltration through the detection of human nasal tissue samples, in vivo and in vitro experiments, so as to further improve the pathogenesis of Eos infiltration in CRSwNP.
慢性鼻窦炎伴鼻息肉（CRSwNP）的复发率与嗜酸性粒细胞（Eos）浸润呈正相关。在CRSwNP患者中，白细胞介素（IL）-19和Eos趋化因子RANTES水平升高已有报道。本研究旨在阐明IL-19在介导RANTES表达和Eos浸润中的作用，特别是在Eos CRSwNP中。方法：从CRSwNP患者和对照组中获取鼻黏膜上皮组织样本，研究组织中IL-19及其受体、嗜酸性粒细胞阳离子蛋白（ECP）和RANTES的表达。培养原代人鼻上皮细胞（HNECs）、成纤维细胞和鼻息肉组织块，然后用IL-19刺激；通过RT-qPCR、ELISA、Western blotting、HE、免疫组化、免疫荧光染色、共聚焦显微镜和Transwell迁移实验检测ERK磷酸化、NF-κB通路激活、RANTES水平、Eos迁移和浸润。建立CRS小鼠模型动物实验检测RANTES的表达。结果：与非Eos CRSwNP和对照组相比，Eos CRSwNP患者鼻黏膜上皮组织中IL-19及其受体（IL-20R1/IL-20R2）、ECP和RANTES的表达显著增加。IL-19与RANTES在鼻黏膜上皮组织中共定位，IL-19显著提高HNEC中RANTES的表达。IL-19抗体和IL-20R1的siRNA降低IL-19对HNECs中RANTES分泌的影响。此外，IL-19诱导的RANTES上调与ERK和NF-κB通路的激活有关。NF-κB的激活是通过ERK通路介导的，在IL-19处理的HNECs中，IL-19增强鼻息肉组织块中Eos的浸润。IL-19可以促进人鼻黏膜上皮组织来源的成纤维细胞分泌RANTES，IL-19可以激活成纤维细胞ERK、NF-κB和JNK通路，ERK、NF-κB和JNK通路抑制剂可以降低IL-19对成纤维细胞中RANTES分泌的影响。在CRS模型中，小鼠鼻腔灌洗液及外周血中RANTES的表达均显著升高。结论：我们的研究结果表明，IL-19通过ERK/NF-κB通路促进HNECs中RANTES的表达，并与Eos CRSwNP患者中Eos的浸润有关。
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