Expression profile of long non-coding RNA in newborn mice with oxygen-induced retinopathy
10.3760/cma.j.issn.1007-9408.2018.01.010
- VernacularTitle:氧诱导新生小鼠视网膜病长链非编码RNA表达谱
- Author:
Fen PENG
1
;
Yu WANG
;
Yunpeng WANG
;
Beibei WANG
;
Xiaoyu ZHOU
;
Suying WU
;
Xiaoguang ZHOU
Author Information
1. 湖北民族学院附属民大医院新生儿科
- Keywords:
Retinopathy of prematurity;
RNA;
long noncoding;
High-throughput nucleotide sequencing;
Mice
- From:
Chinese Journal of Perinatal Medicine
2018;21(1):46-54
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the differences in expression profile of long non-coding RNA (lncRNA) in retina between normal newborn mice and those with oxygen-induced retinopathy (OIR) to lay a foundation for further study of regulatory mechanisms of lncRNA in retinal neovascularization, and to provide a new theoretical basis for the prevention and treatment of retinopathy of prematurity (ROP). Methods A total of 48 cleaning grade C57BL/6J neonatal mice were randomly divided into two groups with 24 in each: OIR group and control group. Those in the OIR group were exposed to hyperoxia [(75±2)％ O2] from 7th to 12th day after birth, and then re-exposed to normoxia (room air) for five days to establish a OIR mouse model. Mice in the control group were raised in room air all the time. Retinal tissues were collected from each mouse on the 17th day after birth. Retinal angiography and hematoxylin-eosin (HE) staining were performed to confirm the establishment of OIR mouse model. Expression profiles of lncRNAs were analyzed by lncRNA high-throughput sequencing and the results were verified by real-time fluorescence quantitative polymerase chain reaction (PCR). Based on the expression of lncRNAs and mRNA and the relationship between their genomic locations, potential target genes of lncRNAs were obtained. Gene ontology analysis (GO) and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway analysis were used for bioinformatics analysis of lncRNA expression profile in OIR. T test and Graphpad prism were used for statistical analysis and spreadsheet. Results Retinal angiography and HE staining identified serious retinal damage in the OIR group. In total, 1118 differentially expressed lncRNAs ( ≥ 2.0-fold difference in expression, P<0.05) relating to OIR between the two groups were identified by lncRNA high-throughput sequencing. Results of real-time fluorescence quantitative PCR confirmed that XLOC_150632 and XLOC_150636 were upregulated in the OIR group as compared with those in the control, while XLOC_122045,XLOC_100454, XLOC_170009 and XLOC_122042 were downregulated. GO analysis showed that 852, 1148 and 2100 target genes of differentially expressed lncRNAs were relating to biological process, cellular component and molecular function, respectively. KEGG analysis showed that the target genes were associated with 36 biological pathways, mainly relating to retinal development, chemotaxis, migration and energy metabolism of vascular endothelial cells. Conclusions lncRNAs are differentially expressed in healthy newborn mice and newborn mice with OIR. Fibroblast growth factor 2 (FGF2), the potential target gene of XLOC_150632 and XLOC_150636, is related to phosphatidylinositol-3-kinase (PI3K) signaling pathway and may be involved in retinal neovascularization. This might be a potential target for ROP prevention and treatment.
