IL-13R130Q基因多态性增强IL-13R信号通路介导气道上皮屏障功能损伤的作用机制研究

The gene-environment interaction(GEI) play an important role in the pathogenesis of asthma. Airway epithelial cell impairment resulted from GEI are the initial event which leading to airway inflammation. Published data indicated that IL-13/IL-13R signaling pathway played an important role in epithelial cells impairment. It is unclear whether IL-13/IL-13R signaling pathway exerted the similar role in airway epithelial cell damage. Our primary study founded that airway epithelial cells also expressed IL-13R, and our previous epidemiological survey confirmed that the incidence of asthma was much higher in individuals with IL-13R130Q gene polymorphism. IL-13R130Q gene polymorphism will lead to enhancing IL-13/IL-13R signaling pathway. Based on those facts, we presume that IL-13R130Q gene polymorphism impaired airway epithelial barrier via enhancing IL-13/IL-13R signaling pathway and thus initiated airway inflammation. To prove our hypothesis, we will evaluate airway epithelial cell impairment and function during IL-13R signaling pathway interfere using transgenic mice with IL-13R130Q gene mutation or siRNA technique both in an OVA-induced asthma model and in vitro. The result from our study will further elucidate the mechanism of airway epithelial cell damage and provide theory evidence for the asthma prevention and treatment.

遗传与环境之间的交互作用是哮喘发病的重要机制。气道上皮细胞是环境因素作用于机体的主要靶细胞。上皮细胞损伤是哮喘过敏性气道炎症的始动环节。IL-13/IL-13R信号通路是参与上皮细胞损伤的重要通路。目前仍未明确IL-13/IL-13R信号通路是否是介导上皮细胞损伤的主要机制。我们前期大样本流行病学调查证实，携带功能增强型IL-13R（IL-13R130Q）的个体哮喘发生率明显升高，且发现气道上皮细胞中表达IL-13R。我们推测IL-13R130Q基因多态性导致IL-13/IL-13R信号通路增强而加重气道上皮细胞损伤是哮喘发病的重要机制。为此，本课题尝试在野生型和携带IL-13R130Q的转基因小鼠中建立OVA诱导的哮喘模型，通过靶向IL-13R的RNA干扰技术手段，观察IL-13R信号通路干预对气道上皮细胞屏障功能及气道过敏性炎症的影响，阐明气道上皮细胞损伤机制，并为预防哮喘提供理论依据

设计针对IL-13Ra1 的 siRNA并转染人支气管上皮细胞，应用IL-13及IL-13R130Q刺激人支气管上皮细胞，结果显示：与野生型IL-13相比，给予IL-13130Q诱导明显的降低了上皮连接蛋白ZO-1、Occludin、E-cadherin 及 ß-catenin 的 mRNA 表达量，上调了上皮细胞p-JAK 及 P-STAT6 的蛋白表达量同时降低了上皮细胞跨膜电阻并增加了上皮通透性；而应用针对 IL-13R a1 siRNA进行干扰，则可以明显抑制il-13及IL-13130Q诱导的上皮链接蛋白ZO-1、Occludin、E-cadherin 及 ß-catenin 的 mRNA 表达量增加，降低人支气管上皮细胞p-JAK 及 P-STAT6 的蛋白表达量，同时增加了上皮细胞跨膜电阻并降低了支气管上皮细胞通透性。以野生型 IL-13 小鼠和 IL-13A2044G（R130Q）转基因小鼠构建屋尘螨（HDM）诱导的气道上皮损伤哮喘小鼠模型，观察并比较两组小鼠一般情况、肺功能、肺部病理结构、上皮链接蛋白表达及支气管肺泡灌洗液中炎症因子表达情况，进一步从体内探索 IL-13130R 野生小鼠和 IL-13130Q 转基因小鼠气道病理生理改变及哮喘症状的改变。

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