Goralska Malgorzata, Fleisher Lloyd N, McGahan M Christine
Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, North Carolina, United States.
Invest Ophthalmol Vis Sci. 2017 Feb 1;58(2):1187-1195. doi: 10.1167/iovs.16-20610.
In humans, vitrectomy is associated with development of nuclear cataracts. Iron catalyzes free radical formation causing oxidative damage, which is implicated in cataract formation. This study was designed to determine if vitreous humor, which can initiate differentiation of lens epithelial cells, would have an effect on iron-handling proteins.
Cultured canine lens epithelial cells were treated with collected canine vitreous humor. Lysates of treated and control cells were separated by SDS-PAGE. Ferritin H- and L-chains, transferrin receptor 1, and aquaporin 0 were immunodetected and quantitated with specific antibodies. Morphologic changes in treated cells were assessed.
Treatment of lens epithelial cells with a 33% (vol/vol) solution of vitreous humor changed the morphology of lens cells and induced expression of aquaporin 0, a marker of fiber cell differentiation that was undetectable in control cells. Treatment did not modify the size of iron-handling proteins but significantly increased content of ferritin from 2.9- to 8.8-fold over control and decreased levels of transferrin receptor by 37% to 59%.
Vitreous humor may significantly limit iron uptake by transferrin/transferrin receptor pathway, and by increasing ferritin levels could profoundly increase the iron-storage capacity of ferritin in lens cells. Vitreous humor may play a significant protective role against iron-catalyzed oxidative damage of lens epithelial cells and therefore in the formation of cataracts.
在人类中,玻璃体切割术与核性白内障的发生有关。铁催化自由基形成,导致氧化损伤,这与白内障的形成有关。本研究旨在确定能够启动晶状体上皮细胞分化的玻璃体液是否会对铁处理蛋白产生影响。
用收集的犬玻璃体液处理培养的犬晶状体上皮细胞。处理后的细胞裂解物和对照细胞裂解物通过SDS-PAGE进行分离。用特异性抗体对铁蛋白H链和L链、转铁蛋白受体1和水通道蛋白0进行免疫检测和定量。评估处理后细胞的形态学变化。
用33%(体积/体积)的玻璃体液溶液处理晶状体上皮细胞改变了晶状体细胞的形态,并诱导了水通道蛋白0的表达,水通道蛋白0是纤维细胞分化的标志物,在对照细胞中无法检测到。处理并未改变铁处理蛋白的大小,但铁蛋白含量显著增加,比对照增加了2.9至8.8倍,转铁蛋白受体水平降低了37%至59%。
玻璃体液可能通过转铁蛋白/转铁蛋白受体途径显著限制铁的摄取,并通过增加铁蛋白水平,可能会显著提高晶状体细胞中铁蛋白的铁储存能力。玻璃体液可能对铁催化的晶状体上皮细胞氧化损伤起到显著的保护作用,因此在白内障的形成中也起到保护作用。