南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (08): 901-.

• •    下一篇

地塞米松对内毒素诱导的小鼠葡萄膜炎的治疗效果及转录组分析

余鹏，邱一果，林茹，符馨予，郝冰涛，雷博   

  • 出版日期:2018-08-20 发布日期:2018-08-20

Retinal transcriptome profile in mice following dexamethasone treatment for endotoxin-induced uveitis

  • Online:2018-08-20 Published:2018-08-20

HTML

PDF

1063

摘要： 目的研究地塞米松（DEX）对内毒素诱导性葡萄膜炎（EIU）的治疗效果及转录组测序探讨其潜在的机制。方法将125 ng 脂多糖( LPS) 注射入雌性BALB/c小鼠玻璃体内建立EIU模型，每隔4 h用0.1% DEX滴眼，共6次。注射LPS后24 h 用裂隙 灯观察小鼠眼前房炎症情况并进行临床评分。苏木精-伊红染色法观察小鼠眼部形态学变化。RNA-seq对EIU模型组和DEX 治疗组小鼠的视网膜进行转录组测序并进行GO和KEGG功能分析。Real-time PCR检测小鼠视网膜炎症细胞因子表达及验证 选出的差异基因。结果连续滴眼6次后，DEX可减轻EIU前房的炎症，并下调炎症因子白介素-6（IL-6）、肿瘤坏死因子-α（TNF- α）、单核细胞趋化蛋白-1（MCP-1）和细胞间黏附分子-1（ICAM-1）mRNA的表达。RNA-seq共检测出52个差异基因，与DEX+ LPS组相比，LPS组中上调的基因有37个，下调的基因有15个。差异基因的功能分析未发现显著富集的GO条目。KEGG富集 分析显示有6个显著上调的KEGG通路和2个显著下调的KEGG通路。KEGG结果提示DEX治疗后可下调RIG-I类受体信号 通路（Ddx58、Ifih1和Isg15）及一些免疫炎症相关基因（Ifit1、H2-T24、Mx2和Eif2ak2）。结论DEX对LPS 引起的小鼠内毒素 诱导性小鼠葡萄膜炎有保护作用，此保护作用可能与下调RIG-I类受体信号通路及一些免疫炎症相关基因有关。

关键词: 转录组测序, 内毒素诱导性葡萄膜炎, RIG-I类受体信号通路, 炎症, 地塞米松

Abstract: Objective To investigate the changes in retinal transcriptome profile of mice with endotoxin-induced uveitis (EIU) following dexamethasone (DEX) treatment and explore the mechanisms underlying the therapeutic effect of DEX. Methods EIU was induced in BALB/c mice by intravitreal injection of 125 ng lipopolysaccharide (LPS), followed by topical application of DEX (0.1%) eye drops every 4 h for 24 h. The anterior chamber inflammation was examined with a slit lamp and the clinical scores were assessed. The morphological changes in the eyes were assessed at 24 h after LPS injection. The retinas were harvested for analysis of transcriptome profile using the next-generation sequencing (NGS)-based RNA sequencing (RNA-seq), and the expressions of the inflammatory cytokines and the differentially expressed genes (DEGs) were verified using real-time PCR. Results DEX alleviated the inflammatory response and reduced the mRNA expressions of IL-6, TNF-α, MCP-1 and ICAM-1 at 24 h after LPS injection. A total of 52 DEGs were identified by RNA-seq. Within these DEGs, 37 genes were upregulated and 15 genes were down-regulated in LPS group as compared with DEX+LPS group. No significantly enriched Gene Ontology (GO) terms was noted. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed 6 up-regulated and 2 down-regulated KEGG pathways. RIG-I-like receptor signaling pathway and several immune- and inflammation-related genes including Ifit1, H2-T24, Mx2 and Eif2ak2 were significantly down regulated by DEX. Verification with RT-PCR yielded results consistent with these findings. Conclusion DEX alleviates LPS-induced inflammatory response in the retina of mice, and such protective effect is probably mediated by RIG-I like receptor signal pathway and the immune-and inflammation-related genes.

Key words: endotoxin-induced uveitis, dexamethasone, transcriptome, RIG-I-like receptor signaling pathway, inflammation