衰老相关LncRNA ANRIL及PINT参与的氧化损伤清除基因GST的表观遗传调控在年龄相关性白内障发病中的作用机制研究

年龄相关性白内障（ARC）病因及确切发病机制尚未探明。我们研究发现氧化损伤清除基因GST表观遗传学改变DNA甲基化、组蛋白修饰与ARC发生相关（已发表），但尚未探讨影响该表观遗传改变的机制。我们预实验发现，衰老相关LncRNA ANRIL及PINT在ARC晶状体中表达降低；结合文献报道提出假设：ANRIL及PINT通过衰老相关Rb/p16和p53/p21通路调控GST表观遗传改变，参与ARC发生。已收集不同类型ARC及透明晶状体，测定衰老相关LncRNA表达；进一步完善体内/体外ANRIL及PINT过表达、Crispr/Cas9模型，观察细胞凋亡、细胞周期；构建氧化损伤/衰老模型，测定抗氧化能力；FISH、RNA pull-down、质谱分析、RIP等探究通路调控机制。揭示衰老相关LncRNA与ARC的关系，从表观遗传学水平探讨ARC发病机制，为ARC防治提供新思路，有助研发抗ARC药物。

The etiology and the exact pathogenesis of the age-related cataract (ARC) remain unclear. Our previous study shows that the epigenetic changes of glutathione S transferase (GST) are associated with ARC formation, but the mechanism of epigenetic changes has not been investigated. Recently, we have observed that there is differential expression of senescence-associated LncRNA ANRIL (antisense non-coding RNA in the INK4 locus) and PINT (p53 induced noncoding transcript) in ARC lens. We hypothesize that ANRIL and PINT can regulate the epigenetic changes of GST by modulating Rb/p16 and p53/p21 pathways during the ARC formation. In current study, we have collected different type of ARC lens and transparent lens to examine the expression of senescence-associated LncRNA. We have also overexpressed ANRIL and PINT in the human lens epithelial cell lines, and we will knockdown and knockout ANRIL and PINT in the same cell lines to investigate whether ANRIL and PINT are related to apoptosis. Flow cytometry will be used to examine the cell cycle. Oxidative damage and aging model both in vitro and in vivo will be constructed to explore whether ANRIL and PINT are involved in oxidative damage and aging. The FISH (fluorescence in situ hybridization), RNA pull-down, mass spectrometry analysis and RNA Binding Protein Immunoprecipitation (RIP) will be used to explore the regulatory mechanism. The study will give us a new insight to elucidate the relationship between the senescence-associated LncRNA and ARC formation and to investigate the pathogenesis of the ARC in epigenetics levels. It will open a new path for early diagnosis of ARC and help to develop potential therapeutic drugs for ARC treatment.