McGahan M C, Grimes A M, Nasisse M P, Fleisher L N
Department of Anatomy, Physiology and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh 27606, USA.
Graefes Arch Clin Exp Ophthalmol. 1995 Jun;233(6):354-9. doi: 10.1007/BF00200484.
Transferrin and Fe concentrations increase in the intraocular fluids in pathological conditions and the lens accumulates Fe during ocular inflammation. Tissues take up Fe from transferrin by two mechanisms, receptor-medicated endocytosis of diferric transferrin and a process occurring at the cell membrane which may be mediated by an oxido-reductase. However, Fe metabolism, transport and storage have not been previously investigated in the lens. This study was designed to characterize the uptake of Fe from transferrin by lens epithelial cells in culture.
Primary, secondary and tertiary cultures of canine lens epithelial cells and cultures obtained from cataractous lenses were studied. Uptake of 59Fe from transferrin by these cultured cells was measured. Transferrin receptor populations were determined in receptor-binding assays.
There was a distinct relationship between the amount of Fe-transferrin added and the amount of Fe taken up, which was linear for the primary cultures but significantly reduced for the secondary, tertiary and cataract cultures (252 +/- 21, 169 +/- 14, 153 +/- 14 and 96 +/- 2 ng Fe/mg protein, respectively). Transferring receptor expression in lens cell cultures was reduced 10-fold within 2 days of addition of serum to cells grown in low-Fe, serum-free medium for 1 week.
The reduction of Fe uptake by the subcultured and cataract cell lines probably reflects a decrease in transferrin receptor expression and in the activity of an alternative pathway for Fe transferrin uptake occurring over time. This reduced Fe uptake may result from long-term exposure to relatively high Fe concentration in the media. A reduction in the expression of the transferrin receptor after incubation with high concentrations of Fe supports this conclusion.
在病理状态下,眼内液中的转铁蛋白和铁浓度会升高,并且在眼部炎症期间晶状体中铁会蓄积。组织通过两种机制从转铁蛋白摄取铁,即双铁转铁蛋白的受体介导的内吞作用以及可能由氧化还原酶介导的发生在细胞膜上的过程。然而,此前尚未对晶状体中的铁代谢、转运和储存进行研究。本研究旨在表征培养的晶状体上皮细胞从转铁蛋白摄取铁的情况。
研究了犬晶状体上皮细胞的原代、传代和三代培养物以及从白内障晶状体获得的培养物。测定了这些培养细胞从转铁蛋白摄取⁵⁹Fe的情况。在受体结合试验中测定转铁蛋白受体数量。
添加的铁 - 转铁蛋白量与摄取的铁量之间存在明显关系,这在原代培养中呈线性,但在传代、三代和白内障培养中显著降低(分别为252±21、169±14、153±14和96±2 ng铁/毫克蛋白)。在低铁、无血清培养基中生长1周的细胞添加血清后2天内,晶状体细胞培养物中转铁蛋白受体表达降低了10倍。
传代培养细胞系和白内障细胞系中铁摄取的减少可能反映了转铁蛋白受体表达的降低以及随着时间推移发生的转铁蛋白摄取替代途径活性的降低。这种铁摄取的减少可能是由于长期暴露于培养基中相对较高的铁浓度。与高浓度铁孵育后转铁蛋白受体表达的降低支持了这一结论。