杯状细胞增生及粘蛋白MUC5AC高表达是哮喘气道粘液高分泌的重要病机，其中hCLCA1基因起了重要的调节作用。白藜芦醇是一种具有抗喘活性的天然化合物，我们在前期哮喘小鼠实验中发现白藜芦醇能显著抑制杯状细胞增生，抑制MUC5AC、 mCLCA3（hCLCA1的异源同种基因）及其上游SPDEF转录因子的表达，因此我们认为白藜芦醇可能通过调节SPDEF/hCLCA1通路抑制杯状细胞增生及MUC5AC表达。为此本课题拟以SPDEF/hCLCA1为研究中心，以人气道上皮细胞为研究对象，观察白藜芦醇对杯状细胞增生、SPDEF、hCLCA1和MUC5AC表达的干预作用，在此基础上通过沉默或过表达SPDEF基因、构建hCLCA1启动子报告载体及EMSA等实验观察SPDEF与hCLCA1的转录调节关系及白藜芦醇的干预作用，最终阐明白藜芦醇调节气道粘液高分泌的分子机制，为白藜芦醇的进一步临床应用奠定实验基础。

Goblet cell hyperplasia and MUC5AC overexpression contribute to mucus hyperproduction in asthma patients, in which hCLCA1 gene plays a critical role. Resveratrol, a polyphenolic stilbene, has been found to exhibit strong anti-inflammatory effects. In our previous studies, we found resveratrol significantly inhibited goblet cell hyperplasia and the expression of MUC5AC, hCLCA1 and the upstream SPDEF transcription factor in a murine model of asthma. Based on these results, we therefore hypothesise that resveratrol might suppress goblet cell hyperplasia and MUC5AC overexpression by regulating SPDEF/hCLCA1 signal pathway. In order to verify the hypothesis, we intend to use lung epithelial cells as the model and to firstly investigate the effect of resveratrol on changes of MUC5AC, hCLCA1 and SPDEF expression level as well as goblet cell hyperplasia. And secondly, we intend to investigate the mechanism of hCLCA1 expresssion regulation by SPDEF and the interference effect of resveratrol by using SPDEF knock down and overexpression cells, hCLCA1 promoter report vector and EMSA. Our study will explain how resveratrol inhibits mucus hypersecretion, which will be helpful for the further pharmaceutical application of resveratrol.

支气管哮喘（简称哮喘）是一种常见慢性疾病，近年来随着空气质量的不断恶化，其发病率呈逐年上升趋势。气道杯状细胞增生及伴随的粘液高分泌和粘蛋白MUC5AC高表达是支气管哮喘的重要病理学改变，抑制粘液高分泌和粘蛋白MUC5AC高表达成了治疗哮喘的重要目标之一。本项目原计划通过相关研究工作阐明SPDEF转录因子对hCLCA1-MUC5AC轴的调控模式，同时揭示白藜芦醇调节气道粘液高分泌和粘蛋白MUC5AC表达的分子机制。3年来课题组按照计划任务书要求，以MUC5AC和白藜芦醇为研究中心，通过哮喘小鼠实验发现白藜芦醇干预能显著缓解哮喘小鼠杯状细胞增生和气道粘液高分泌，进一步的基因和蛋白表达分析显示白藜芦醇显著抑制了哮喘小鼠粘蛋白MUC5AC、CLCA3（人CLCA1的异源同种基因）和SPDEF的表达水平。体外细胞实验也证明白藜芦醇可以抑制SPDEF、CLCA1和MUC5AC的表达，并且白藜芦醇以SPDEF依赖的方式调控MUC5AC的表达，靶向SPDEF是白藜芦醇抑制气道粘液高分泌和粘蛋MUC5AC表达的新途径。在此基础上我们通过启动子分析揭示了SPDEF对CLCA1的转录调控模式，通过信号通路分析揭示了白藜芦醇抑制STAT6和AKT信号通路从而下调SPDEF表达的分子机制。另外我们发现白藜芦醇能抑制哮喘小鼠气道重塑及炎症因子（IL-6）表达，其作用机制可能与调控PTEN和SIRT1相关。最终通过3年的研究工作，课题组取得了一些具有自主创新的科研成果，累计发表受本项目资助的文章7篇（SCI论文4篇），揭示了白藜芦醇干预哮喘气道粘液高分泌和炎症反应的分子机制，为白藜芦醇的进一步抗哮喘治疗应用提供了实验数据和理论支持。