胰岛素对人RPE细胞增殖及转化生长因子-β_2分泌的影响

Effects of insulin on proliferation and secretion of transforming growth factor-β_2 of human retinal pigment epithelial cells

目的:探讨胰岛素对RPE细胞增殖和分泌转化生长因子-β2(TGF-β2)的影响。方法:将不同浓度的胰岛素(0.1～10×103U/mL)加入培养基刺激ARPE-19细胞不同时间(0～48h)。采用MTS法检测RPE细胞的增殖情况;采用ELISA法检测各种处理情况下RPE细胞分泌TGF-β2的情况,Real-time PCR法检测TGF-β2mRNA的表达情况。结果:MTS结果显示,作用12h后,各实验组与对照组比较RPE细胞明显增殖(P<0.05)。ELISA结果表明胰岛素可明显促进RPE细胞分泌TGF-β2,并呈时间及剂量依赖性。Real-time PCR结果显示,胰岛素作用24h后,TGF-β2mRNA表达上调(P<0.01),且10×103U/mL胰岛素组TGF-β2mRNA表达明显高于0.1×103U/mL胰岛素组(P<0.01)。结论:RPE细胞是眼内近视信号因子TGF-β2的重要来源,胰岛素可以通过促进RPE细胞增殖并分泌TGF-β2进行信号传递,促进近视发生。

AIM： To investigate if insulin can promote retinal pigment epithelial （RPE） cell proliferation and the secretion of transforming growth factor 62（TGF-β2）. METHODS： Human RPE cells were exposed to different concentrations insulin at different times （0 to 48 hours）. We used the MTS method to test the proliferation of RPE cells. And we used the ELISA method to text the secretion of TGF-152 of RPE cells in various cases, and the Real-time PCR method to test the expressions of TGF-β2 mRNA. RESULTS： The MTS results showed that after 12 hours exposure of insulin RPE cells proliferated significantly （P〈 0.05）. ELISA results showed that insulin can significantly promote the secretion of TGF-β2 of RPE cells, and this process was independent of time and dose. And Real- time PCR results showed that 24 hours after the stimulation of insulin, TGF-β2 mRNA increased significantly （ P〈0.01 ）, and 10 × 103 U/mL insulin group, TGF-β2 mRNA expression were significantly higher than the 0. 1 × 103 U/mL insulin group （ P〈0.01 ）. CONCLUSION： RPE cells are important intraocular sources of TGF-β2, and insulin can promote the proliferation of RPE cells and the secretion of TGF-β2 to promote myopia.