LE540对视网膜色素上皮 bFGF及TGF-β2的影响

摘要

目的:观察全反式视黄酸β受体阻断剂LE540对培养的豚鼠视网膜色素上皮细胞(RPE)分泌碱性成纤维细胞生长因子(bFGF)及转化生长因子-β2(TGF-β2)的影响.方法:胰蛋白酶消化法培养原代豚鼠RPE细胞,角蛋白免疫组织化学染色鉴定为阳性后,传2代细胞后用于实验.不同实验组分别加入10×10-6mol/L的全反式视黄酸ATRA和5×10-6mol/L的LE540,设试剂对照.ELISA检测ATRA及LE540作用2h、4h、6h、8h、16h后TGF-β2和bFGF的分泌量.结果:ELISA结果显示:(1)加入ATRA后,实验组TGF-β2的分泌量与对照组比较明显增加,而bFGF的分泌量明显减少(P＜0.05).(2)加入LE540后,实验组较对照组RPE分泌TGF-β2均明显减少(P＜0.05),差别有统计学意义.同时bFGF的分泌量没有明显变化(P=0.178).结论:全反式视黄酸能通过视黄酸β受体途径促进视网膜色素上皮细胞分泌TGF-β2,同时通过其他途径能抑制bFGF的分泌.提示有可能通过阻断视黄酸β受体来控制近视的进展.%Objective: To evaluate the roles of retinoic acid receptor beta antagonist ,LE540, on secretion of basic fibroblast growth factor(bFGF) and transfer growth factor-β2 (TGF-β2) in cultured retinal pigment epithelial cells (RPE). Methods: Retinal pigment epithelial cells were digested by the trypsin. RPE cells were identified by keratin and the second generation were used in experiment. 10×10-6 mol/L of all-trans retinoic acid ATRA and 5 × 10-6 mol / L of LE540 were respectively added to different experimental group.The same dose of medium was used in control group.The bFGF and TGF-β2 secreted by RPE cells were detected using ELISA method at 2、4、6、8、16 hours after the addition of ATRA and LE540. Results: Cultured cells showed the positive response for keratin. ELISA results showed that:( 1 )the secretion of TGF-β2 was significantly increased after action of ATRA in comparison with control group, while the secretion of bFGF significantly reduced (P ＜0.05). (2)the secretion of TGF-β2 was significantly reduced after action of LE540 in comparison with control group (P ＜0.05). While the secretion of bFGF did not significantly change (P＝0.178). Conclusion: The all-trans retinoic acid can promote the secretion of TGF-β2 mediated by retinoic acid receptor- β, and while inhibit the secretion of bFGF through other way. Our data sugges that it possible to control the progress of myopia by blocking the retinoic acid receptor β.