Long non-coding RNA histone deacetylase 4 antisense RNA 1 (HDAC4-AS1) inhibits HDAC4 expression in human ARPE-19 cells with hypoxic stress

Abstract

Age-related macular degeneration (AMD) is resulted from choroidal neovascularization (CNV)mediated cicatrization and vision loss. The sustained retinal hypoxia in retinal pigment epithelium (RPE) cells was reported to contribute to CNV. However, the underlying genetic regulatory network of hypoxia response in RPE is not fully understood. In this study, human ARPE-19 RPE cells were cultured under the anoxia for 24 h and later re-oxygenated in normoxia. Then the transcriptome was investigated via high throughput sequencing. We observed that long noncoding RNA (lncRNA) histone deacetylase 4 antisense RNA 1 (HDAC4-AS1) was increased in hypoxic condition compared to normal control and decreased after re-oxygenation addition, while the change of HDAC4 expression was reduced in hypoxic condition compared to normal control and up-regulated after re-oxygenation addition in ARPE-19 cells. Furthermore, HDAC4-AS1 knockdown could suppress the transcription activity of HDAC4 only in hypoxia condition, and fluorescence in situ hybridization and pull down assay indicated that transcripts of HDAC4-AS1 could substantially bind to the promoter of HDAC4 and facilitate the recruitment of HIF-1α. Finally, we also determined the specific regions of HDAC4-AS1 that contribute to the interaction with HIF-1α and the promoter of HDAC4. Taken together, these outcomes declared that HDAC4-AS1 could inhibit HDAC4 expression through regulating HIF-1α in human ARPE-19 cells with hypoxic stress.

Investigation on the expression regulation of RIPK1/RIPK3 in the retinal ganglion cells (RGCs) cultured in high glucose

“…As a consequence, a series of changes occur in the intracellular environment inducing cell death. As a high metabolic organ, the retina consumes a large amount of oxygen in the human body, so the retina is very sensitive to ischemia and hypoxia [ 24 , 25 ]. In a recent study, Du et al used high-glucose DMEM to establish a retinal ischemia/reperfusion (I/R) injury model in retinal ganglion cells.…”

Long non-coding RNA H19 protects against intracerebral hemorrhage injuries via regulating microRNA-106b-5p/acyl-CoA synthetase long chain family member 4 axis

“…All primers were designed and synthesized by Nanjing Genscript Biotech Co., Ltd. (Jiangsu, China), and GAPDH and U6 were used as internal references. -Fold changes in the indicated genes were calculated using the 2 −ΔΔCt method [ 24 ]. The primer sequences were as follows:…”

Long non-coding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) regulates fibroblast growth factor receptor substrate 2 (FRS2) by targeting microRNA (miR)-29-3p in hypertrophic scar fibroblasts

“…HSFs were isolated from the clinical samples and were incubated in DMEM containing 10% FBS and 1% penicillin/streptomycin (Gibco, Califonia, USA) under 5% CO 2 at 37°C [ 24 ].…”

“…After transfection, the cells were plated into 96-well plates and cultured for 24 h [ 24 ]. Thereafter, CCK-8 reagents (20 µL/well) were added, and the cells were cultured for another 4 h. The absorbance values at 450 nm were detected using a SpectraMax M5 microplate reader.…”