GSK3B/β-catenin/TCF通路介导Pirfenidone调控人眼Tenon's囊成纤维细胞周期的机制研究

Abstract: The Excessive proliferation of Human Tenon's capsule fibroblasts(HTFs) is the direct and major cause of the scarring which always appear after glaucoma surgery. Studies have shown that Pirfenidone(PFD) can effectively block the cell proliferation cycle of HTFs and keep them stay in G1 phase with an unknown mechanism. GSK3B/β-catenin/TCF signaling pathways is a major pathway that regulate cell survival and does an important role on the inhibition of PFD to TGF-β/Smad. GSK3B widely expressed in HTFs and the increased activity of GSK3B pathways weakened the cell proliferation cycle of HTFs and indicated that PFD may regulate the cell proliferation cycle though GSK3B/β-catenin/TCF pathway. Our research intend to clarify the mechanism of the effect and regulation of PFD to the cell proliferation cycle of HTFs though GSK3B/β-catenin/TCF pathway. Our research will provide new ideas and therapeutic targets to the scarring after glaucoma surgery.

人Tenon's囊成纤维细胞过度增殖是青光眼手术滤过区过度瘢痕化主要原因。研究表明Pirfenidone能有效阻滞人Tenon's囊成纤维细胞于G1期，GSK3B/β-catenin/TCF信号通路是调控细胞生存的重要通路，对Pirfenidone抑制TGF-β/Smad有调控作用。已发现GSK3B在人Tenon's囊成纤维细胞广泛表达，而GSK3B通路活性增强可阻遏人Tenon's囊成纤维细胞周期，提示Pirfenidone可能通过GSK3B/β-catenin/TCF通路进行细胞周期调控。本课题拟明确：GSK3B//β-catenin/TCF通路介导的Pirfenidone在人Tenon's囊成纤维细胞的作用及具体调控机制。本研究将为青光眼术后滤过区瘢痕化调控提供新的思路和治疗靶点。

人Tenon's囊成纤维细胞过度增殖是青光眼手术滤过区过度瘢痕化主要原因。我们前期研究表明Pirfenidone能有效阻滞人Tenon's囊成纤维细胞于G1期，鉴于瘢痕化调控是涉及多靶点、多通路的复杂过程，细胞周期调控可能是抗过度瘢痕化的新靶点。GSK3B、AKT和MAPK等信号通路是调控细胞生存的重要通路，本研究通过Pirfenidone的G1阻滞与这些通路的关系，希望厘清Pirfenidone细胞周期调控作用在抗过度瘢痕化中的作用和机制。研究结果表明Pirfenidone通过抑制CDK4/6-cyclinD和CDK2-cyclinE复合体的活性调控G1/S检测点，并通过AKT、ERK1/2、JNK和p38信号通路，下调CDK2、CDK6、cyclinD1、cyclinD3和cyclinE的表达，阻滞HTFs于G1期；在动物实验中，经过前期研究的筛选，以PTMC-F127-PTMC水凝胶作为本研究PFD眼用剂型的载体，进行体内效应试验和药代动力学研究，Pirfenidone/PTMC-F127-PTMC眼部应用能显著提高Pirfenidone的生物利用度。同时，Pirfenidone是一个多靶点药物，在瘢痕调控过程中有明显的抑制新生血管化作用。本研究为青光眼术后滤过区瘢痕化调控提供新的药物和治疗靶点。