烟曲霉感染相关microRNAs在肺泡巨噬细胞炎症反应通路中的作用研究

项目名称： 烟曲霉感染相关microRNAs在肺泡巨噬细胞炎症反应通路中的作用研究

项目编号： No.81200004

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 邵宏涛

作者单位： 中国人民解放军南京军区南京总医院

项目金额： 23万元

中文摘要： 平衡促炎/抗炎反应以避免机体二次损伤是侵袭性曲霉感染治疗成功的重要因素。肺泡巨噬细胞（AMs）在此过程中起免疫调节作用。但如何通过负反馈通路调控信号转导，抑制AMs产生过多促炎因子的具体机制尚不清楚。我们前期研究发现烟曲霉（AP）感染的小鼠AMs中miR-146a、155表达上调，并伴有IL-1β、TNF-a升高，提示这两种miRNA可能参与对AMs胞内信号转导通路的调控。本项目拟建立AP感染小鼠AMs模型，通过miRNA芯片、免疫印迹和qPCR技术，筛选并鉴定烟曲霉感染诱导的miRNA；通过生物信息学预测和报告载体实验鉴定其靶基因；分别阻断NF-κB通路蛋白后测定miRNA及IL-1β、TNF-a表达；并通过miRNA过表达和抑制实验分析其负性调控功能。探讨AP感染、miRNA变化对信号转导通路的作用机制，对进一步阐明曲霉感染的分子致病机制具有重要意义。

中文关键词： 烟曲霉；小RNA；免疫反应；巨噬细胞；

英文摘要： The balance between pro- and anti-inflammation plays an important role in treating invasive Aspergillus fungi infection. In addition to executing its primary phagocytic function upon excountering the AP, AMs are thought to play the pivotal role in coordinating the host inflammatory response. However，the mechanism of how to avoid over-production of pro-inflammatory cytokines in AMs is still unknown. We have found previously that the expression of miR-146a、155 have increased in mouse alveoli marcrophages infected by aspergillus conidia, and the corresponding oligonucleotide restrained the expression of IL-1β、TNF-a, which indicated that the two miRNAs may involved in the regulation of signal transduction in AMs. Thus we plan to establish the aspergillus infected cell model, screening and evaluating the corresponding miRNA induced by aspergillus infection through miRNA assay, northern blotting and qPCR, predicting the target gene, assaying the expression of miRNA,IL-1β and TNF-a after blocking the NF-κB signal pathway, analyzing the negative regulation of the target miRNA through over-expression and inhibition of the corresponding miRNA.We try to investigate the mechanism of how the target miRNA regulate the signal transduction,then clarifying further the mechanism molecularly in aspergillus infection.

英文关键词： Aspergillus；miRNA；immulogy；macrophage；