Liu Hongling, Feng Guangzhong, Wu Lan, Fu Shaoying, Liu Ping, Yang Wei, Zhang Xiaomei
Department of Ophthalmology, The 1st Affiliated Hospital, Harbin Medical University, Harbin, PR China.
Mol Vis. 2010 Aug 16;16:1646-53.
The objective of the present study was to investigate the efficacy of rapamycin on rabbit lens epithelial cell proliferation, migration, and secretion of extracellular matrix fibronectin (Fn).
Rabbit lens epithelium cells (rLECs) were isolated from 1 month old rabbit. rLECs were either cultured for 24, 48, or 72 h with different doses of rapamycin (0.1, 1, and 10 ng/ml). The proliferation kinetics, proliferating cell nuclear antigen (PCNA) expression, and basic fibroblast growth factor (bFGF)-induced migration of rLEC was determined by methyl thiazol tetrazolium (MTT) assay, western blotting and transwell chamber assay, respectively. The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence.
Rapamycin significantly inhibited rLEC proliferation and PCNA protein expression when administered doses and time periods except for 0.1 ng/ml for 24 h. bFGF-induced migration rLECs was inhibited by pretreatment with rapamycin for 48 h. Extracellular matrix Fn formation of rLECs was also reduced by rapamycin.
In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation. Thus, rapamycin may have a potential inhibition of posterior capsule opacification (PCO) and needs further study.
本研究的目的是探讨雷帕霉素对兔晶状体上皮细胞增殖、迁移及细胞外基质纤连蛋白（Fn）分泌的影响。
从1月龄兔分离兔晶状体上皮细胞（rLECs）。将rLECs分别用不同剂量的雷帕霉素（0.1、1和10 ng/ml）培养24、48或72小时。分别采用甲基噻唑基四氮唑（MTT）法、蛋白质印迹法和Transwell小室法测定rLEC的增殖动力学、增殖细胞核抗原（PCNA）表达及碱性成纤维细胞生长因子（bFGF）诱导的迁移。通过免疫荧光检测雷帕霉素对Fn合成的影响。
除24小时0.1 ng/ml外，在给药剂量和时间段内，雷帕霉素均显著抑制rLEC增殖和PCNA蛋白表达。雷帕霉素预处理48小时可抑制bFGF诱导的rLECs迁移。雷帕霉素还可减少rLECs细胞外基质Fn的形成。
在我们的研究中，雷帕霉素强烈抑制rLEC增殖、bFGF诱导的迁移及细胞外基质Fn的形成。因此，雷帕霉素可能对后囊膜混浊（PCO）具有潜在抑制作用，有待进一步研究。
