Liu Wei, Ma Chao, Li Hao-Yu, Chen Lan, Yuan Shan-Si, Li Kai-Jun
Department of Ophthalmology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.
Exp Ther Med. 2020 Nov;20(5):38. doi: 10.3892/etm.2020.9165. Epub 2020 Sep 1.
The present study aimed to investigate the effect of microRNA (miR)-146a on the secretion of hyaluronic acid (HA) and collagen I in Graves' ophthalmopathy (GO) orbital fibroblasts, and identify potential novel targets for the clinical treatment of GO. Orbital fibroblasts were extracted from orbital connective tissue, and primary cells were identified via immunohistochemistry. The levels of HA and collagen I in orbital fibroblasts of non-GO controls and patients with GO were examined via reverse transcription-quantitative PCR (RT-qPCR). miR-146a was overexpressed or inhibited in primary orbital fibroblasts via lentiviral infection, and the levels of HA and collagen I following miR-146a overexpression or inhibition were detected via ELISA and RT-qPCR. The results indicated that the mRNA expression of HA and collagen I was higher in orbital fibroblasts from patients with GO compared with the non-GO cohort. Overexpression of miR-146a reduced, and inhibition of miR-146a increased the production of HA and collagen I in GO orbital fibroblasts. In conclusion, it was demonstrated that miR-146a downregulated the secretion of HA and collagen I in GO orbital fibroblasts , which may affect glycosaminoglycan aggregation and collagen deposition in GO.
本研究旨在探讨微小RNA(miR)-146a对格雷夫斯眼病(GO)眼眶成纤维细胞中透明质酸(HA)和I型胶原蛋白分泌的影响,并确定GO临床治疗的潜在新靶点。从眼眶结缔组织中提取眼眶成纤维细胞,并通过免疫组织化学鉴定原代细胞。通过逆转录定量PCR(RT-qPCR)检测非GO对照组和GO患者眼眶成纤维细胞中HA和I型胶原蛋白的水平。通过慢病毒感染在原代眼眶成纤维细胞中过表达或抑制miR-146a,并通过酶联免疫吸附测定(ELISA)和RT-qPCR检测miR-146a过表达或抑制后HA和I型胶原蛋白的水平。结果表明,与非GO队列相比,GO患者眼眶成纤维细胞中HA和I型胶原蛋白的mRNA表达更高。miR-146a的过表达降低了,而miR-146a的抑制增加了GO眼眶成纤维细胞中HA和I型胶原蛋白的产生。总之,证明了miR-146a下调了GO眼眶成纤维细胞中HA和I型胶原蛋白的分泌,这可能影响GO中糖胺聚糖聚集和胶原蛋白沉积。