Department of Ophthalmology, Soonchunhyang University Bucheon Hospital, Soonchunhyang University College of Medicine, Bucheon, Republic of Korea.
Department of Ophthalmology, Severance Hospital, The Institute of Vision Research, Yonsei University College of Medicine, Seoul, Republic of Korea.
PLoS One. 2019 Aug 15;14(8):e0221077. doi: 10.1371/journal.pone.0221077. eCollection 2019.
To investigate the role of microRNA (miR)-27a and miR-27b in adipogenesis in an in vitro model of Graves' orbitopathy (GO).
Orbital fat tissues were harvested from GO and non-GO participants for primary orbital fibroblast cultures. The expression levels of miR-27a and miR-27b between GO and non-GO orbital fat tissues were compared. During adipogenesis of GO orbital fibroblasts, the expression levels of miR-27a and miR-27b were determined, and the effects of mimics of miR-27a and miR-27b transfection on adipogenesis of GO orbital fibroblast were investigated.
Real time-polymerase chain reaction showed significantly more decreases in miR-27a and miR-27b levels in orbital fat tissues in GO participants than in non-GO participants (p < 0.05). The expression of both miR-27a and miR-27b was highest in orbital fibroblasts at day 0 and declined gradually after the induction of adipogenic differentiation. The expression levels of PPARγ, CCAAT/enhancer binding protein (C/EBP)α and C/EBPβ were decreased and Oil Red O-stained lipid droplets were lower in GO orbital fibroblasts transfected with miR-27a and miR-27b mimics than in negative controls.
Our results indicated that miR-27a and miR-27b inhibited adipogenesis in orbital fibroblasts from GO patients. Further studies are required to examine the potential of miR-27a and miR-27b as targets for therapeutic strategies.
为了在 Graves 眼病(GO)的体外模型中研究 microRNA(miR)-27a 和 miR-27b 在脂肪生成中的作用。
从 GO 和非 GO 参与者中采集眼眶脂肪组织用于原代眼眶成纤维细胞培养。比较 GO 和非 GO 眼眶脂肪组织中 miR-27a 和 miR-27b 的表达水平。在 GO 眼眶成纤维细胞的脂肪生成过程中,确定 miR-27a 和 miR-27b 的表达水平,并研究 miR-27a 和 miR-27b 模拟物转染对 GO 眼眶成纤维细胞脂肪生成的影响。
实时聚合酶链反应显示 GO 参与者眼眶脂肪组织中 miR-27a 和 miR-27b 的水平明显低于非 GO 参与者(p<0.05)。在诱导成脂分化后,miR-27a 和 miR-27b 的表达在第 0 天的眼眶成纤维细胞中最高,并逐渐下降。GO 眼眶成纤维细胞中转染 miR-27a 和 miR-27b 模拟物后,PPARγ、CCAAT/增强子结合蛋白(C/EBP)α 和 C/EBPβ 的表达降低,油红 O 染色的脂滴减少。
我们的结果表明 miR-27a 和 miR-27b 抑制了 GO 患者眼眶成纤维细胞中的脂肪生成。需要进一步研究以检查 miR-27a 和 miR-27b 作为治疗策略靶点的潜力。