背景:近视是全世界儿童和青少年最常见的眼病之一,巩膜重塑在近视发展中起作用。然而,导致近视相关巩膜重塑的起始因素和信号通路的鉴定尚不清楚.本研究旨在鉴定巩膜重塑的生物标志物,以阐明近视的发病机制。
方法:使用基因表达综合(GEO)和比较毒性基因组学数据库(CTD)挖掘来鉴定与近视巩膜重塑相关的miRNA-mRNA调控网络。实时定量PCR(RT-qPCR),蛋白质印迹,免疫荧光,H&E染色,Masson染色,流式细胞术检测FOXO信号通路的变化,纤维化,凋亡,细胞周期,巩膜重塑的其他相关因素。
结果:miR-15b-5p/miR-379-3p可以调节FOXO信号通路。证实研究证实,眼轴长度显著增加,巩膜厚度较薄,miR-15b-5p的水平,miR-379-3p,PTEN,p-PTEN,FOXO3a,细胞周期蛋白依赖性激酶(CDK)抑制剂1B(CDKN1B)增加,LIM组IGF1R水平降低。CDK2,细胞周期蛋白D1(CCND1),通过流式细胞术评估细胞周期阻滞表明近视巩膜中G1/S细胞周期停滞。BAX水平的升高和BCL-2水平的降低表明近视巩膜的凋亡增强。此外,我们发现转化生长因子-β1(TGF-β1)的水平,胶原蛋白1型(COL-1),α-平滑肌肌动蛋白(α-SMA)减少,提示近视眼发生巩膜重塑。
结论:miR-15b-5p/miR-379-3p可通过IGF1R/PTEN/FOXO信号通路调控巩膜细胞周期和凋亡,从而促进近视进展中巩膜重塑。
BACKGROUND: Myopia is one of the most common eye diseases in children and adolescents worldwide, and scleral remodeling plays a role in myopia progression. However, the identity of the initiating factors and signaling pathways that induce myopia-associated scleral remodeling is still unclear. This study aimed to identify biomarkers of scleral remodeling to elucidate the pathogenesis of myopia.
METHODS: The gene expression omnibus (GEO) and comparative toxicogenomics database (CTD) mining were used to identify the miRNA-mRNA regulatory network related to scleral remodeling in myopia. Real-time quantitative PCR (RT-qPCR), Western blot, immunofluorescence, H&E staining, Masson staining, and flow cytometry were used to detect the changes in the FOXO signaling pathway, fibrosis, apoptosis, cell cycle, and other related factors in scleral remodeling.
RESULTS: miR-15b-5p/miR-379-3p can regulate the FOXO signaling pathway. Confirmatory studies confirmed that the axial length of the eye was significantly increased, the scleral thickness was thinner, the levels of miR-15b-5p, miR-379-3p, PTEN, p-PTEN, FOXO3a, cyclin-dependent kinase (CDK) inhibitor 1B (CDKN1B) were increased, and the levels of IGF1R were decreased in Len-induced myopia (LIM) group. CDK2, cyclin D1 (CCND1), and cell cycle block assessed by flow cytometry indicated G1/S cell cycle arrest in myopic
sclera. The increase in BAX level and the decrease in BCL-2 level indicated enhanced apoptosis of the myopic
sclera. In addition, we found that the levels of transforming growth factor-β1 (TGF-β1), collagen type 1 (COL-1), and α-smooth muscle actin (α-SMA) were decreased, suggesting scleral remodeling occurred in myopia.
CONCLUSIONS: miR-15b-5p/miR-379-3p can regulate the scleral cell cycle and apoptosis through the IGF1R/PTEN/FOXO signaling pathway, thereby promoting scleral remodeling in myopia progression.