LE540对视网膜色素上皮bFGF及TGF-β_2的影响

Effects of LE540 on secretion of bFGF and TGF-β_2 of RPE cells in vitro

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-β(2TGF-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10×10-6 mol/L的全反式视黄酸ATRA和5×10-6 mol/L的LE540,设试剂对照。ELISA检测ATRA及LE540作用2 h、4 h、6 h、8 h、16 h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.