光控开关定量调控鼻息肉鼻黏膜上皮细胞Ras/Raf/MEK/ERK信号通路功能的研究

光遗传学技术将光作为可调式和非侵入式介质动态调控细胞信号通路，时空分辨率高、可定量重复、操作简单。因而，近年其与疾病的研究得到广泛认可与推进。鼻息肉是鼻科常见病，鼻黏膜上皮细胞Ras/Raf/MEK/ERK信号通路参与其发病机制已被揭示，本研究拟利用光遗传学技术定量调控鼻息肉鼻黏膜上皮细胞的该信号通路，通过① Western blot、RT-PCT、MTT检测上皮细EGFR、Ras等因子确定鼻息肉上皮细胞ERK信号通路的抑制。②分别质粒转染蓝光和红光调控开关CRY2-CIBN和PhyB-PIF，在上皮细胞中建立光控元件，Western blot、RT-PCT、MTT、ELISA检测调控后鼻黏膜增殖，确定光控定量范围。③将筛选出的光控开关动物体内试验，观察利用光遗传学方法对鼻黏膜上皮ERK信号通路的功能定量调控，继而干预鼻息肉的发生发展。以期为鼻息肉的精准治疗开拓可能的光遗传学治疗靶点。

Optogenetic approach to quantitative control of intracellular signaling by its unique advatages, such as non destruction, non-invasion, high temporal and spatial resolution and high reversibility. Ras/Raf/MEK/ERK pathway plays an important role during the nasal polyps epithelial cells remodeling. This project is based upon the optogentic approach, plans on utilizing nasal epithelial cells by optogenetics to control the Ras/Raf/MEK/ERK pathway in nasal polyps. Concrete research content include: ①analyse the inhibiting phenomenon of nasal epithelial cells Ras/Raf/MEK/ERK pathway in nasal polyps by Western blot、RT-PCT、MTT.②construct light-gated protein-protein CRY2-CIBN and PhyB-PIF interaction system respectively to regulate the activation pattern of the Ras/Raf/MEK/ERK pathway in nasal epithelial cells of nasal polyps.③nasal polyps animal models will be performed to test whether optogentics control nasal epithelial cells could intervene the progression of nasal polyps. The object is construct optogentic-gate to control biological activity of nasal epithelial cells,focusing on those that might have potential clinical therapeutic target in preventing nasal polyps.