Differentially expressed genes analysis of retinal vascular endothelial cells under hyperglycemia and hyperlipidemia induced by α-melanocyte-stimulating hormone

Objective

To investigate the effect of α-melanocyte-stimulating hormone(α-MSH) on the expression of mRNA and long noncoding RNA (lncRNA) in retinal vascular endothelial cells stimulated by hyperglycemia and hyperlipidemia.

Methods

The simian retinal vascular endothelial cells (RF/6A)were cultured and divided into normal control group, model control group, 0.1 μmol/L α-MSH group, 0.5 μmol/L α-MSH group and 1.0 μmol/L α-MSH group.The cells were stained with CM-H₂DCFDA to detect cell antioxidant capacity.The optimal concentration of α-MSH was screened.The cells from normal control group, model control group and α-MSH treatment group were collected at 24 hours after treatment, the total RNA was extracted, the cDNA library was constructed, and the high throughput RNA sequencing (RNA-seq) was carried out with bioinformatics analysis to analyze the expression profiling of mRNA and lncRNA.

Results

The fluorescence intensity of cells in 0.5 μmol/L α-MSH group was significantly lower than that in model control group (P<0.05). α-MSH of 0.5 μmol/L was chosen as the optimal concentration for subsequent experiments.Compared with the model control group, 243 mRNAs were significantly down-regulated, while 81 mRNAs were up-regulated in the α-MSH treatment group; 53 lncRNAs were markedly up-regulated and 6 lncRNAs were down-regulated in the α-MSH treatment group.Bioinformatics analysis showed that the major enrichment pathways of the down-regulated genes were transforming growth factor-β(TGF-β) signaling pathway, focal adhesion signaling pathway and extracellular matrix (ECM) receptor interactions pathways, and the main biological process involved was the regulation of small GTPase-mediated signal transduction.The co-expression gene enrichment pathways of differentially expressed lncRNA included ECM receptor interaction and hypoxia inducible factor-1(HIF-1) pathway, et al.These pathways were mainly involved in the biological processes, such as axon guidance and positive regulation of transcription from RNA polymerase Ⅱ promoter.

Conclusions

Under hyperglycemia and hyperlipidemia, the influence of α-MSH on the transcriptome of the retinal vascular endothelial cells manifests the downregulation of mRNA and upregulation of lncRNA.α-MSH may upregulate the lncRNA expression, which downregulates the downstream mRNA expression.