长链非编码RNA BANCR在人晶体混浊发生的作用及莱菔硫烷的保护机制

Age-related cataract, which is characterized by lens opacity, is the main disease leading to the elderly visually impaired. The anti-oxidative and autophonic defense ability plays an important role in its occurrence, but the mechanism and effective prevention are out of research. Our early work has confirmed that low concentrations of sulforaphane (SFN) can prevent lesn opacity, which upregulates the Nrf2 antioxidant pathway. LncRNA BANCR is up-regulated as ceRNA of miR-176a which increases the expression of ERK on MAPK pathway. It is speculated that the occurrence of human turbidity and the protective mechanism of SFN are related to the double regulation of antioxidant pathway and autophagy pathway. The two passages resist the oxidative damage by crosstalk. The study investigates the dynamic changes of autophagy and the effect of SFN on autophagy in human lens epithelial cells under oxidative stress, and to explore the role of lncRNA BANCR as an interaction factor on Nrf2 antioxidant pathway and MAPK autophagy pathway. This study will help to understand the pathogenesis of cataract from the dynamic balance of cell microenvironment and the integration of information pathway, and provide a new target for drug control.

以晶体混浊为特征的年龄相关性白内障是导致老年视力障碍的主要疾病，抗氧化和自噬防御能力减退对其发生起重要作用，但具体作用机制不详亦缺少有效的防治手段。我们前期工作已经证实氧化应激下长链非编码RNA BANCR作为ceRNA结合miR-176a下调MAPK通路上自噬调节蛋白ERK表达；而莱菔硫烷可上调长链非编码RNA BANCR同时上调Nrf2抗氧化通路预防晶体混浊。因此推测，人晶体混浊的发生和莱菔硫烷的保护机制涉及抗氧化通路和自噬通路的联合调节，由长链非编码RNA BANCR关联两通路发生crosstalk，共同抵御氧化损害。本课题拟研究氧化压力下人晶体上皮细胞自噬水平的变化及莱菔硫烷对其的调节；并探索长链非编码RNA BANCR作为交互因子对Nrf2抗氧化通路及MAPK自噬通路的作用。本课题将从细胞微环境动态平衡和信息通路整合的角度深入理解白内障发病机制，提供能联合多系统的新药物靶点

后发性白内障是白内障术后最常见的并发症，也是导致患者视力再次下降的最主要的原因。PCO源于内囊袋中残留晶状体上皮细胞在后囊膜上增殖，迁移，转分化为分泌胶原和基底膜养物质使后囊膜在此浑浊，上皮-间质转化被认为是形成PCO主要病理机制。本研究探索长链非编码RNA BANCR（lncRNA BANCR） 在人晶状体上皮细胞FHL24中对上皮-间质转化的作用，并进一步探究了其调控晶体上皮细胞增殖、凋亡及自噬的作用及相关分子机制。运用qReal - time PCR 检测TGF-β诱导对FHL24细胞内EMT相关标志物α-SMA，E-cadherin，Coll I ，ZO1及BANCR mRNA相对表达量。细胞中转染BANCR。Western 印迹检测各组细胞中EMT相关标志蛋白及LC3Ⅱ/ Ⅰ的蛋白表达。MTT 法检测各组细胞的增殖，凋亡情况。与正常对照组比较，TGF-β诱导组细胞中BANCR, α-SMA, Coll I， ZO1 mRNA的相对表达量明显增加, 而E-cadherin mRNA显著下降, 差异均有统计学意义 (t=-5.031, -7.145, -9.023, -6.012, 5.097均P < 0.05)，以上因子的蛋白表达趋势相同，差异均有统计学意义（均P < 0.05)。siRNA-BANCR TGF-β诱导组细胞中E-cadherin mRNA相对表达量比 siRNA TGF-β诱导组显著增加(t= -9.98, p<0.05）；α-SMA，Coll I 及ZO1 mRNA相对表则显著减少（t= 9.003; 27.738; 19.620, p<0.05）。抑制BANCR后细胞增殖活力48、72 h 时细胞活性显著降低(t= 5.032, 9.041，均p<0.05），细胞凋亡率显著升(t=16.772，p<0.001）。自噬标志蛋白LC3 - II/ LC3 - I 比例增加 (P <0.05)。综上长链非编码BANCR参与了晶体上皮细胞的抑制其上皮-间质转化，抑制BANCR可抑制晶体上皮细胞增殖、增加凋亡及自噬的发生。以期通过本研究从细胞微环境动态平衡和信息通路整合的角度加深对PCO 发病机制的理解，为实现多防御系统联合的药物防治提供新靶点。