节点文献
PI3K/Akt信号通路参与血小板源性生长因子促进人晶状体上皮细胞移行
Involvement of the PI3K/Akt Signaling Pathway in Platelet-Derived Growth Factor-Induced Migration of Human Lens Epithelial Cells
【作者】 熊炜；
【导师】 唐罗生；
【作者基本信息】 中南大学 ， 临床医学， 2012， 博士
【摘要】 后发性白内障指白内障囊外摘除术后,或外伤性白内障部分皮质吸收后所形成的晶状体后囊膜混浊(PCO),是白内障囊外摘除术后影响视力最常见的并发症。有报道,成人白内障术后2年内,有20%-30%的患者因后发障而再度失明,5年内有43%的患者因后发障而行Nd:YAG激光后囊膜切开术。后发性白内障的组织病理学研究表明白内障术后残留的晶状体上皮细胞的增殖、迁移、纤维化生是形成后发障的主要原因,目前有很多此方面的研究,以寻求在这方面找到好的预防和治疗PCO的方法。目的：晶状体上皮细胞的移行在PCO的形成中起到很重要的作用。目前,其移行的机理还远未阐明。我们研究的目的是探讨PDGF刺激晶状体上皮细胞引起其移行的信号通路和下游分子。方法：用离体的刮伤愈合实验和跨膜移行实验来研究晶状体上皮细胞的移行。用蛋白印迹分析来研究PDGF刺激HLE-B3细胞对PDGFRβ, PI3K/Akt信号通路和MAPK信号通路活化的影响；对细胞突起分子表达的影响；是否有稳定β-catenin的作用。用免疫荧光分析来研究β-catenin在细胞内分布位置的变化。结果：PDGF促进HLE-B3细胞的移行,PI3K/Akt信号通路参与其中。通过PDGF/PDGFRβ轴向激活的PI3K/Akt信号通路可以上调细胞突起分子的表达和稳定且改变β-catenin在细胞内的分布,这是是PDGF促进细胞移行的机理之一。结论：本研究直接证明PI3K/Akt信号通路与人晶状体上皮细胞的移行有关,提供了寻找治疗PCO的新的方法的思路。
【Abstract】 PURPOSE Posterior capsular opacification (PCO) is caused partially by the migration of lens epithelial cells. To date, the mechanism of the migration is largely unknown. The purpose of this study was to investigate the effect of platelet-derived growth factor (PDGF)-triggered signaling pathways and its downstream effectors in the migration of lens epithelial cells.METHODS In vitro scratch-wound healing and transwell migration assays were used to measure the migration of lens epithelial cells. The activation of PDGFRβ, phosphatidylinositol3-kinase (PI3K)/protein kinase B (Akt) and mitogen activation protein kinase (MAPK) pathways, the impact of PDGF treatment on the expression of cell protrusion molecules, and the stabilization of β-catenin were measured by western blotting. The translocation of β-catenin was detected using indirect immunofluorescence.RESULTS PDGF was found to enhance cell migration, which depended on the PI3K/Akt pathway. The activation of the PI3K/Akt pathway by the PDGF/PDGFRβ axis induced the upregulation of cell protrusion molecules and stabilization and translocation of β-catenin, contributing to enhanced cell migration. CONCLUSION Data from this study directly linked the central PI3K/Akt pathway to lens epithelial cell migration and pointed to new avenues for therapeutic intervention in PCO.
【Key words】 Posterior capsular opacification； PDGF； migration oflens epithelial cell； PI3K/Akt pathway；