基于SPDEF/hCLCA1通路探讨白藜芦醇抑制气道粘液高分泌的分子机制

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.