Department of Ophthalmology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, China.
Medical School of Nantong University, Nantong, Jiangsu, China.
Sci Rep. 2024 Nov 9;14(1):27359. doi: 10.1038/s41598-024-78479-z.
Abnormal growth of blood vessels (choroidal neovascularization) can lead to age-related macular degeneration (AMD) and eventually cause vision loss due to detachment of the retinal pigmented epithelium. This indicates that choroidal neovascularization is important for the treatment of AMD. The circadian clock in the mammalian retina is responsible for controlling various functions of the retina, enabling it to adjust to changes in light and darkness. Recent studies have revealed a potential connection between the circadian clock and eye diseases, although a cause-and-effect relationship has not been definitively established. C57BL/6J male mice (aged 6 weeks) were randomly divided into two groups (Control group: 9:00-21:00 light period (300 lx); Jet lag group: 8-hour phase advance once every 4 days). A laser-induced CNV model was created after 2 weeks of feeding in a controlled or jet-lagged environment. Then, full transcriptome sequencing was performed. The pathways regulated by differentially expressed mRNAs were identified by GO analysis and GSEA. Further protein networks were constructed with the STRING database and Cytoscape software. WGCNA was used to further explore the co-expression modules of these differential genes and the correlation between these differential genes and phenotypes. ceRNA networks were constructed with miRanda and TargetScan. The pathways associated with the overlapping differentially expressed mRNAs in the ceRNA network were identified, and the hub genes were validated by qPCR. A total of 661 important DEGs, 31 differentially expressed miRNAs, 106 differentially expressed lncRNAs and 87 differentially expressed circRNAs were identified. GO and GSEA showed that the upregulated DEGs were mainly involved in reproductive structure development and reproductive system development. The STRING database and Cytoscape were used to determine the protein interaction relationships of these DEGs. WGCNA divided the expression of these genes into several modules and screened the hub genes of each module separately. Furthermore, a ceRNA network was constructed. GO analysis and GSEA showed that these target DEmRNAs mainly function in wound healing, cell spreading, epiboly involved in wound healing, epiboly, and morphogenesis of an epithelial sheet. Finally, ten key genes were identified, and their expression patterns were confirmed by real-time qPCR. In this study, we investigated the regulatory mechanism of ceRNAs in choroidal neovascularization according to different light-dark cycles in the eyeball.
异常血管生长（脉络膜新生血管）可导致年龄相关性黄斑变性（AMD），并最终导致视网膜色素上皮脱离引起视力丧失。这表明脉络膜新生血管对于 AMD 的治疗很重要。哺乳动物视网膜中的生物钟负责控制视网膜的各种功能，使其能够适应光暗变化。最近的研究揭示了生物钟与眼部疾病之间的潜在联系，尽管尚未确定因果关系。将 6 周龄 C57BL/6J 雄性小鼠（雄性）随机分为两组（对照组：300 lx 的 9:00-21:00 光照期；时差组：每 4 天提前 8 小时相位）。在受控或时差环境中喂养 2 周后，建立激光诱导的 CNV 模型。然后进行全转录组测序。通过 GO 分析和 GSEA 鉴定差异表达 mRNA 调控的途径。使用 STRING 数据库和 Cytoscape 软件构建进一步的蛋白质网络。使用 WGCNA 进一步探索这些差异基因的共表达模块以及这些差异基因与表型之间的相关性。使用 miRanda 和 TargetScan 构建 ceRNA 网络。鉴定 ceRNA 网络中重叠差异表达 mRNA 相关的途径，并通过 qPCR 验证枢纽基因。共鉴定出 661 个重要的差异表达基因、31 个差异表达 miRNA、106 个差异表达 lncRNA 和 87 个差异表达 circRNA。GO 和 GSEA 表明，上调的差异表达基因主要参与生殖结构发育和生殖系统发育。STRING 数据库和 Cytoscape 用于确定这些 DEG 的蛋白质相互作用关系。WGCNA 将这些基因的表达分为几个模块，并分别筛选每个模块的枢纽基因。此外，构建了 ceRNA 网络。GO 分析和 GSEA 表明，这些靶标 DEmRNAs 主要在伤口愈合、细胞扩散、涉及伤口愈合的外包、外包和上皮片的形态发生中发挥作用。最后，确定了十个关键基因，并通过实时 qPCR 验证了其表达模式。在这项研究中，我们根据眼球内不同的光暗周期，研究了 ceRNA 在脉络膜新生血管中的调控机制。
