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lncRNA H19 sponging miR-93 to regulate inflammation in retinal epithelial cells under hyperglycemia via XBP1s

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Abstract

Objective

Diabetic retinopathy (DR) is a major complication of both type 1 and type 2 diabetes. Recently, inflammation was found to play an important role in DR pathogenesis. But the mechanism has not been fully understood.

Methods

ARPE-19 cells were cultured under normal condition and high-glucose condition, then the expressions of miR-93, XBP1s and lncRNA H19 were measured using RT-qPCR or western blots. Besides, the mRNA level of eIF2α and GRP78 and protein level of p-eIF2α and GRP78 were measured by RT-qPCR or western blots. In addition, RT-qPCR and ELISA were adopted to examine the expression and secretion of cytokine factors in these conditions. Dual-luciferase reporter gene assay was used to elucidate the binding and regulation among XBP1s, miR-93 and H19. RNA immunoprecipitation was also performed to verify the interaction between H19 and miR-93. The expressions of DNAJC3 and DNAJB9, the downstream targets of XBP1s, were detected by RT-qPCR.

Results

We identified that H19 and XBP1s were down-regulated in ARPE-19 cells under high-glucose condition, while miR-93 was up-regulated. ER stress inducers TM and IRE1 inhibitor STF-083010 were adopted and data suggest that ER stress could be induced during high-glucose treatment. In addition, the altered expressions of miR-93, XBP1s and H19 might mediate the increased level of pro-inflammatory cytokines. Furthermore, miR-93 interacted with either lncRNA H19 or XBP1s then modulating the inflammatory processes.

Conclusions

H19 played an important role in regulating inflammatory processes in retinal endothelial cells under high-glucose condition through modulating miR-93/XBP1s axis.

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Availability of data and material

All data generated or analyzed during this study are included in this published article.

Abbreviations

DR:

Diabetic retinopathy

miR:

MicroRNA

XBP1s:

Spliced X-box-binding protein 1

lncRNA:

Long non-coding RNA

qPCR:

Quantitative polymerase chain reaction

ELISA:

Enzyme-linked immunosorbent assays

ER:

Endoplasmic reticulum

TNF-α:

Tumor necrosis factor-α

VEGF:

Vascular endothelial growth factor

RECs:

Retinal endothelial cells

MIAT:

Myocardial infarction-associated transcript

ANRIL:

Antisense RNA to INK4 locus

RIP:

RNA-binding protein immunoprecipitation

LPS:

Lipopolysaccharides

UPR:

Unfolded protein response

IRE1α:

Inositol-requiring protein-1α

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Correspondence to Rong Luo.

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Luo, R., Xiao, F., Wang, P. et al. lncRNA H19 sponging miR-93 to regulate inflammation in retinal epithelial cells under hyperglycemia via XBP1s. Inflamm. Res. 69, 255–265 (2020). https://doi.org/10.1007/s00011-019-01312-1

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Keywords

  • Long non-coding RNA
  • H19
  • MicroRNA
  • miR-93
  • Diabetic retinopathy
  • Inflammation