氯离子通道阻滞剂NPPB在青光眼滤过泡瘢痕化中的作用及机制研究

滤过性手术是治疗青光眼的主要手术方式，但术后5年滤过泡存活率不超过50%。其失败的原因是结膜成纤维细胞（HconF）通过分化、增殖、迁移、ECM合成及胶原收缩导致滤过泡瘢痕形成。研究表明氯离子通道阻滞剂NPPB抑制多种细胞增殖、迁移和细胞周期进展，目前还没有通过阻滞氯离子通道而抗滤过泡瘢痕化的研究。我们前期研究发现CLC-2氯离子通道通过PI3K/Akt信号通路调控HconF细胞纤维化过程。本课题将在前期工作的基础上，拟采用分子生物学、流式细胞仪、迁移等实验方法研究NPPB在HconF细胞纤维化中作用及其机制；建立SD大鼠青光眼滤过手术模型，观察CLC-2氯离子通道的表达；用siRNA-CLC-2和NPPB抑制大鼠结膜CLC-2氯离子通道的表达，检测各项瘢痕化指标，阐明CLC-2氯离子通道在青光眼滤过泡瘢痕化中的作用，为青光眼抗滤过泡瘢痕化治疗提供新靶点。

Filtration surgery is the main operation method to treat glaucoma, but the 5 years survival rate of filtering bleb is less than 50%. involved in the filtering bleb scarring process of glaucoma filtration surgery by Conjunctival fibroblasts differentiation，proliferation, migration, extracellular matrix (ECM) synthesis and collagen contraction which lead to filtering bleb scarring is the main main cause of surgery failure. Our previous studies have confirmed that CLC-2 chloride channel regulated the differentiation, proliferation, migration, extracellular matrix (ECM) of HconF via PI3K/Akt signaling pathways. A number of studies have found that chloride channel blocker NPPB involved in the process of cell proliferation, migration and cell cycle progression in a variety of cell type. In this topic various methods (Molecular biology, flow cytometry, cell wound scratch assay, Transwell migration assay method) would be used to investigate the effect of NPPB on differentiatio, proliferation, migration, ECM synthesis and collagen contraction of human conjunctival fibroblasts. Then we would build rat glaucoma filtration surgery model to analyze the effect of CLC-2 siRNA and NPPB on scarring processes of filtering bleb after glaucoma filtration surgery. And further explore the signaling pathways by which NPPB regulate the scarring processes of filtering bleb to provide new targets for anti filtering bleb scarring.