雷公藤多苷对溃疡性结肠炎黏膜愈合中紧密连接及其miRNAs调控机制的研究

Tripterygium wilfordii polyglycosidium (TWP) has been widely used to treat autoimmune diseases. However, few researches have been focused on its therapeutic mechanism upon ulcerative colitis (UC). The goal for UC treatment is mucosa healing and mucosa healing quality has close correlation with tight junction (TJ). Previous study shows that TWP can control UC inflammation and promote mucosa healing. Besides, certain miRNAs can regulate TJ through certain target genes mediated by certain pathways. Finally, we speculate that TWP can promote mucosa healing through regulating TJ, the mechanism of which mainly based on interfering certain miRNA mediated pathways. With such foundation, in order to study the mechanism of TWP towards TJ, experiment is carried out on cell and animal level. With the help of miRNA transfection and knock-down technology, Luciferase assay will be performed to verify and identify the corresponding relations between candidate miRNAs and their target genes. Over expression of certain miRNAs in cells is performed to verify the correlation between miRNA and distribution of TJ-related protein. Target gene agonist is also performed to demonstrate the regulatory effect of target gene and its mediated pathway towards TJ. This project is going to study the mechanisom of TWP, eventually to reveal the molecular basis of TWP towards mucosa healing and TJ, which can provide new ideas for optimization of clinical plan and new drug research.

雷公藤多苷（TWP)广泛用于治疗自身免疫疾病，但TWP治疗溃疡性结肠炎（UC）的机制研究甚少。UC治疗目标是黏膜愈合，而粘膜愈合质量与紧密连接（TJ）状况密切相关。前期研究表明TWP能控制UC炎症、具有黏膜愈合作用；还发现若干miRNAs可能通过调控某些靶基因介导的信号通路调节TJ。我们推测：针对UC，TWP是通过调节TJ起到黏膜愈合作用，干预这些miRNAs的调控途径是TWP调节TJ的一种作用机制。基于此，我们从细胞及动物两个层面，开展TWP对肠上皮TJ作用研究；在细胞中针对miRNA调控，结合荧光素酶报告基因实验验证miRNA与靶基因之间的对应关系；细胞中过表达miRNA验证miRNA与TJ表达分布的关系；利用靶基因拮抗剂验证靶基因介导信号通路对TJ的调节，以研究TWP对TJ的作用机制，从而揭示TWP黏膜愈合作用的分子基础及其作用机制，为临床方案优化及新药研法提供重要数据。

雷公藤多苷（TWP）广泛用于治疗自身免疫疾病，但TWP治疗溃疡性结肠炎（UC）的机制研究甚少。UC治疗目标是黏膜愈合，而粘膜愈合质量与紧密连接（TJ）状况密切相关。前期研究表明TWP能控制UC炎症、具有黏膜愈合作用；还发现若干miRNAs可能通过调控某些靶基因介导的信号通路调节TJ。基于此，本项目从细胞及动物两个层面，开展TWP对肠上皮TJ作用研究；并在细胞中调控miRNA，结合荧光素酶报告基因实验验证miRNA与靶基因之间的对应关系；细胞中过表达miRNA验证miRNA与TJ表达分布的关系；利用靶基因拮抗剂验证靶基因介导信号通路对TJ的调节，以研究TWP对TJ的作用机制，从而揭示TWP黏膜愈合作用的分子基础及其作用机制，为临床方案优化及新药研法提供重要数据。