Epithelial-Mesenchymal Transition-Like Phenotypic Changes of Retinal Pigment Epithelium Induced by TGF-β Are Prevented by PPAR-γ Agonists
Epithelial-Mesenchymal Transition-Like Phenotypic Changes of Retinal Pigment Epithelium Induced by TGF-β Are Prevented by PPAR-γ Agonists
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DOI：
10.1167/iovs.12-10488
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发表时间：
2012-10-01
影响因子：
4.4
通讯作者：
Kinoshita, Shigeru
中科院分区：
文献类型：
--
作者：
Hatanaka, Hiroki;Koizumi, Noriko;Kinoshita, Shigeru
目的.视网膜色素上皮细胞（retinal pigment epithelial cells，RPEC）的纤维化改变是导致增殖性玻璃体视网膜病变（proliferative vitreoretinopathy）和增殖性糖尿病视网膜病变（proliferative diabetic retinopathy）等增殖性眼病的部分原因。本研究的目的是检测过氧化物酶体增殖物激活受体-γ（PPAR-gamma）激动剂对灵长类RPEC纤维化改变的影响。对从食蟹猴眼分离的猴RPEC（MRPEC）进行传代培养。为了诱导纤维化变化，将MRPEC与TGF-β 2（3 ng/mL）一起培养，并且还在TGF-β 2和PPAR-γ激动剂吡格列酮（30 μ M）的共存下培养。通过相差显微镜和免疫细胞化学分析评估培养的MRPEC的表型。Western blot检测Smad 2/Smad 3蛋白的磷酸化水平。原代MRPEC培养为单层六边形细胞，ZO-1、Na+/K+-ATP酶和RPE 65均呈阳性表达。存在吡格列酮时，细胞形态和这些标志物的表达得以维持，而不存在吡格列酮时，细胞延长，这些标志物的表达降低。相反，鬼笔环肽、α-平滑肌肌动蛋白和纤连蛋白的表达在吡格列酮存在下减少，而在不存在下增加。Western blot结果显示吡格列酮可抑制Smad 2/Smad 3蛋白的磷酸化。PPAR-gamma激动剂吡格列酮通过抑制TGF-β信号传导抑制原发性MRPEC的纤维化变化。吡格列酮可能是一种临床上适用的有效的药物治疗增殖性眼病。（Invest Ophthalmol维斯科学。2012;53：6955-6963）DOI：10.1167/iovs.12-10488
PURPOSE. Proliferative eye diseases, such as proliferative vitreoretinopathy and proliferative diabetic retinopathy, are caused partly by fibrotic change of retinal pigment epithelial cells (RPECs). The purpose of our study was to examine the effect of the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist on the fibrotic change of primate RPECs.METHODS. Monkey RPECs (MRPECs) isolated from a cynomolgus monkey eye were subcultured. To induce fibrotic change, MRPECs were cultured with TGF-beta 2 (3 ng/mL), and also cultured in the coexistence of TGF-beta 2 and the PPAR-gamma agonist pioglitazone (30 mu M). The phenotype of the cultured MRPECs was evaluated by phase contrast microscopy and immunocytochemical analysis. The phosphorylation of Smad2/Smad3 proteins was examined by Western blot analysis.RESULTS. Primary MRPECs were cultured as a monolayer with a hexagonal cell shape, and positive expression of ZO-1, Na+/K+-ATPase, and RPE65 was confirmed. Cell morphology and the expression of these markers were maintained in the presence of pioglitazone, whereas the cells were elongated and the expression of these markers was reduced in its absence. Conversely, the expression of phalloidin, alpha-smooth muscle actin, and fibronectin was reduced in the presence of pioglitazone, whereas it was increased in the absence. Western blot assay demonstrated that phosphorylation of Smad2/Smad3 proteins was suppressed by pioglitazone.CONCLUSIONS. The PPAR-gamma agonist pioglitazone inhibited the fibrotic change of primary MRPECs through the suppression of TGF-beta signaling. Pioglitazone might prove to be a clinically applicable and effective pharmaceutic treatment for proliferative eye diseases. (Invest Ophthalmol Vis Sci. 2012;53:6955-6963) DOI:10.1167/iovs.12-10488