磁珠诱导的慢性高眼压性青光眼小鼠模型转录组分析

Transcriptomic analysis in a mouse model of magnetic beads-induced chronic intraocular hypertension glaucoma

目的:利用转录组测序(RNA-seq)和生物信息学技术对磁珠诱导的慢性高眼压性青光眼小鼠模型的视网膜差异表达基因(DEGs)及信号通路的变化进行分析,探讨慢性高眼压青光眼视网膜损伤的生物学机制。方法:4~6周龄雄性C57BL/6J小鼠共6只,随机分为实验组、对照组。实验组通过2次对小鼠单眼前房注射磁珠悬液,阻断房水流出通道,模拟慢性高眼压性青光眼,对照组未行处理。取第48天实验组造模眼和对照组小鼠单眼的视网膜,进行RNA-seq和生物信息学分析,并进一步分析DEGs和线粒体共表达基因。结果:在注射2次磁珠后小鼠眼压升高并维持在较高水平。共筛选出606个DEGs,经GO和KEGG通路分析主要富集在能量代谢相关通路。GSEA分析表明氧化磷酸化、糖酵解通路下调。进一步分析发现DEGs和线粒体基因重叠共59个,富集在质子跨膜转运蛋白活性、线粒体蛋白复合物、氨基酸降解等方面。PPI网络分析揭示Uqcrfs1、Cyc1和Atp5f1a等核心基因。结论:RNA-seq和生物信息学分析提示能量代谢失调参与慢性高眼压性青光眼的视网膜损伤和退行性变,线粒体功能障碍导致轴突转运失调可能在青光眼的发病中发挥重要作用。

Objective:To analyze the alterations in retinal differential expression genes(DEGs)and signaling pathways in a mouse model of chronic intraocular hypertension glaucoma induced by magnetic beads,and to utilize transcriptomic sequencing(RNA-seq)and bioinformatics to elucidate the biological mechanism of retinal injury.Methods:Six male C57BL/6J mice,aged 4-6 weeks,were randomly assigned to either the experimental or control group.The experimental group received two injections of magnetic bead suspension into the anterior chamber of a single eye to obstruct the aqueous humor outflow channel,simulating chronic intraocular pressure glaucoma,while the control group received no treatment.RNA-seq and bioinformatics analyses were conducted on the retinas of the experimental and control groups on the 48th day,followed by further analysis of differentially ex-pressed genes(DEGs)and mitochondrial co-expression genes.Results:Following two injections of magnetic beads,mice exhibited increased and sustained intraocular pressure.A total of 606 DEGs were identified in the retinas of mice between the experimental and control groups.GO functional analysis and KEGG pathway analysis of DEGs revealed enrichment in energy metabolism.GSEA analysis showed that oxidative phosphorylation and glycolysis pathways were down-regulated.Fur-ther analysis showed that there were 59 overlapping DEGs and mitochondrial genes,which were en-riched in proton transmembrane transporter activity,mitochondrial protein complex,amino acid degra-dation and so on.PPI network analysis highlighted hub genes as Uqcrfs1,Cyc1,and Atp5f1a.Con-clusion:RNA-seq and bioinformatics analysis suggest energy metabolism disorders play a crucial role in retinal damage and degeneration in chronic hypertensic glaucoma.Furthermore,mitochondrial dys-function,leading to dysregulation in axonal transport,may be a significant factor in the occurrence and development of glaucoma.