Role of microRNA-146a in regulation of fibrosis in orbital fibroblasts from patients with Graves' orbitopathy
Role of microRNA-146a in regulation of fibrosis in orbital fibroblasts from patients with Graves' orbitopathy
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DOI：
10.1136/bjophthalmol-2017-310723
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发表时间：
2018-03-01
影响因子：
4.1
通讯作者：
Yoon, Jin Sook
中科院分区：
文献类型：
--
作者：
Jang, Sun Young;Park, Seong Jun;Yoon, Jin Sook
目的探讨microRNA-146 a（miR-146 a）在Graves' orbitopathy（GO）体外模型中对纤维化的调控作用。使用定量实时PCR分析了GO和非GO眼眶成纤维细胞中转化生长因子（TGF-β）（一种促进纤维化的强效细胞因子）对miR-146 a表达的影响。通过蛋白质印迹分析在GO眼眶成纤维细胞中检查过表达的miR-146 a对TGF-诱导的纤维化标志物的影响。结果TGF-β 1可诱导GO患者眼眶成纤维细胞中miR-146 a表达增加，并呈时间和浓度依赖性。miR-146 a模拟物进一步降低TGF-诱导的纤连蛋白、胶原蛋白I和β-平滑肌肌动蛋白的产生。与对照组相比，miR-146 a模拟物显著降低了Smad 4和TRAF 6蛋白水平，而与对照组相比，miR-146 a抑制剂显著增加了Smad 4和TRAF 6蛋白水平。结论miR-146 a在TGF-β诱导的纤维化标志物的产生中起负调节作用。因此，miR-146 a可能参与调节GO患者眼眶成纤维细胞中的纤维化。
Aim To examine the role of microRNA-146a (miR-146a) in the regulation of fibrosis in an in vitro model of Graves' orbitopathy (GO).Methods Orbital fat/connective tissues were harvested from patients with GO and non-GO for primary orbital fibroblast cultures. The effects of transforming growth factor- (TGF-), a potent cytokine that promotes fibrosis, on miR-146a expression were analysed in GO and non-GO orbital fibroblasts using quantitative real-time PCR. The effects of overexpressed miR-146a on TGF--induced fibrotic markers were examined in GO orbital fibroblasts by western blot analysis. Expression ofSma and Mad related family (Smad) 4/tumour necrosis factor receptor-associated factor 6 (TRAF6) after transfection of miR-146a mimics or inhibitors were examined.Results TGF- induced an increase in miR-146a expression in orbital fibroblasts from patients with GO in a time-dependent and concentration-dependent manner. miR-146a mimics further decreased the production of TGF--induced fibronectin, collagen I and -smooth muscle actin protein. The Smad4 and TRAF6 protein levels were significantly decreased by miR-146a mimics, compared with control mimics, and significantly increased on inhibition of miR-146a production compared with a control.Conclusions miR-146a plays a role as a negative regulator in the production of TGF--induced fibrotic markers. Thus, miR-146a may be involved in the regulation of fibrosis in orbital fibroblasts from patients with GO.