血管心外膜活性物质(BVES)在哮喘导致的气道上皮损伤中的作用研究

As an evolutionarily conserved transmembrane protein, blood vessel epidemal substance (BVES) is postulated to play a role in cell adhesion and cell motility. Emerging evidence indicates that this protein is present in muscle and epithelia of developing embryos and the adult. However, the expression, function and potential roles of BVES in epithelial injury are not completely understood. To clarify the mechanism underlying epithelial injury of asthmatic airways, first, the acute and chronic asthma model on E3 rat will be established, and the expression of BVES in different stages of asthma will be screened by real-time quantification PCR, western blotting and immunofluorescence. The relationship between BVES expression level and severity of airway epithelium injury will be evaluated after BVES upregulation or downregulation in vivo. Meanwhile,the samples from asthma patients will be used to test and verify BVES level and asthmaic indexes. SSH will be used to detect differential expression genes in high or low stable expression of BVES in epithelial cell. Then, The target gene of BVES in the processes of airway epithelium injury will be defined. Finally, different cytokines or transcription factors will be used to stimulate primary airway epithelium or epithelial cell line, and the expression of BVES will be screened to find upstream factors which regulate the BVES expression. In this project, we will reveal the BEVS function in epithelial injury in acute and chronic asthma, provide the significant theoretical basis for the research of asthmatic pathogenesis, and offer new targets for the medicine research and development.

为了阐明紧密连接蛋白血管心外膜活性物质（BVES）在哮喘气道上皮损伤过程中的作用机制，本研究首先建立E3大鼠的气道致敏模型、急性哮喘模型和慢性哮喘模型，采用RT-PCR、蛋白免疫印迹、免疫荧光等技术检测BVES在哮喘发病不同阶段的表达；并而在动物模型中上调或下调BVES表达水平后观察上皮损伤相关指标的改变，随后在病人的标本中进行验证，以明确BVES的表达与气道上皮损伤的关系；继而在气道上皮细胞中，构建BVES稳转系，采用抑制性消减杂交寻找其下游差异表达的基因，同时分析已知的BVES下游信号通路，以揭示BVES调控气道上皮损伤的分子机制；筛选可影响气道上皮细胞中BVES表达的刺激因子，并结合已知的信号通路和DNA甲基化水平进行分析，阐明调控BVES表达的上游机制。通过该研究不仅可阐明BVES在哮喘发病过程中调节气道上皮损伤的分子基础，而且为哮喘的防治提供实验基础和理论依据。

血管心外膜活性物质（BVES）可调整上皮的结构与上皮细胞的生物学活动。本项目研究了血管心外膜活性物质（BVES）对气道上皮细胞功能的调节，及其对过敏性哮喘的影响。在小鼠哮喘模型中BVES的表达下降。上调BVES在小鼠气道的表达水平会增强气道炎症水平和气道重塑，而下调BVES在小鼠气道的表达水平会减轻气道炎症水平和气道重塑。改变BVES的水平可影响上皮细胞的基因表达。上调BVES的表达可下调A549细胞表面趋化因子受体CCR3和CCR4的表达，下调BVES的表达可上调TGF-β1的表达。EGF和TGF-β1联合刺激可上调BVES的表达。