Toll样受体（TLR）在机械通气肺损伤（VILI）中发挥重要作用。我们前期研究证明了VILI的产生有赖于TLR4/MyD88通路，新近通过全基因组关联分析发现WISP1是VILI的易感基因。在巨噬细胞中我们证明：一方面WISP1能显著放大TLR通路的促炎效应，另一方面TLR4介导了WISP1的表达/作用。因此本研究聚焦"WISP1-TLR4正反馈促炎通路"：首先比较VILI敏感鼠和耐受鼠机械通气后WISP1与TLRx表达水平差别；其次从动物和细胞水平证明WISP1表达/作用与TLR4及相关（CD14、MyD88或TRIF）通路的关系；再采用免疫共沉淀、293细胞TLR转染测试和激光共聚焦筛选和证实WISP1与TLRx的分子作用机制。最后使用巨噬细胞TLR4基因敲除小鼠和WISP1干预实验，阐明WISP1在VILI中作用途径。本研究有利于理解和开发通过阻断"正反馈"环节的肺损伤防治新策略。

Recent studies show Toll-like receptors (TLR) in innate immunity play an important role in Ventilator-induced lung injury (VILI). Our previous work had showed TLR4/MyD88 pathway contributed to VILI. We recently using Genome-wide association study (GWAS) with 23 inbred mouse strains in VILI model identified that WNT1 inducible signaling pathway protein 1 (Wisp1) gene as one of significant single nucleotide polymorphisms associated with VILI. We verified the synergistic effect between extracelluar matrix protein WISP1 and TLR signaling in macrophages. Further, we found TLR4 signaling pathway is involved in the production and effect of WISP1. The current study focuses on the role of " WISP1-TLR4 Positive feedback Loop" in VILI: First, we will observe the differences of WISP1 and TLR expression in mechanical ventilation model using VILI-sensitive and resistent mice. Second, we will verify the relationship between the production and effect of WISP1 and TLR4 associated pathway (including CD14, TLR4, MyD88 and TRIF) in vivo and in vitro. Third, we need to further confirm the interplay between WISP1 and TLR by co-immunoprecipitation, TLR transfection test in HEK293 cells and confocal microscopy. Last, we use macrophage-TLR4 knock out mice and WISP1 protein inhalation to clarify the mechanism of WISP1 in VILI. The project will help understand and explore the novel treatment for acute lung injury through blocking the " WISP1-TLR4 Positive feedback Loop" .