Effect of targeted inhibition of miR-628-5p by long non-coding RNA KCNQ1 overlapping transcript 1 on high glucose-induced retinal pigment epithelial cell injury
10.13389/j.cnki.rao.2024.0117
- VernacularTitle:长链非编码RNA(lncRNA)KCNQ1重叠转录物1靶向抑制miR-628-5p在高糖诱导的视网膜色素上皮细胞损伤中的作用
- Author:
Chaojun QIN
1
;
Xian WANG
Author Information
1. 550025 贵州省贵阳市,贵州医科大学临床医学院
- Keywords:
long non-coding RNA KCNQ1 overlapping transcript 1;
miR-628-5p;
high glucose;
retinal pigment epithe-lium;
apoptosis;
oxidative stress
- From:
Recent Advances in Ophthalmology
2024;44(8):613-618
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of targeted inhibition of miR-628-5p by long non-coding RNA(lncRNA)KCNQ1 overlapping transcript 1(KCNQ1OT1)on high glucose(HG)-induced retinal pigment epithelial cell injury.Meth-ods The real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was performed to detect KCNQ1OT1 expression in serum and aqueous humor of patients with diabetic retinopathy(DR),type 2 diabetes mellitus(T2DM)and uveitis.Human retinal pigment epithelial cells(ARPE-19)were treated with different concentrations of glucose,and their KCNQ1OT1 expression was detected to determine the optimal glucose concentration.ARPE-19 cells were divided into the control(CON)group,HG group,HG+si-NC group,HG+si-KCNQ1OT1 group,HG+miR-NC group,HG+miR-628-5p group,HG+si-KCNQ1OT1+anti-miR-NC group,and HG+si-KCNQ1OT1+anti-miR-628-5p group.The expression levels of KCNQ1OT1 and miR-628-5p in cells were measured by RT-qPCR.The cell apoptosis was detected by flow cytometry.The malondialdehyde(MDA)level and superoxide dismutase(SOD)activity were detected by enzyme-linked immunosorbent assay.The expression level of cleaved Caspase-3 protein was measured by Western blot.The targeting relationship between KCNQ1OT1 and miR-628-5p was detected by dual-luciferase reporter assay.Results The expression levels of KCNQ1OT1 in serum and aqueous humor of DR patients were higher than those of T2DM patients and uveitis patients,and those of uve-itis patients were the lowest(all P＜0.05).Compared with the 0 mmol·L-1 glucose group,the expression levels of KC-NQ1OT1 in ARPE-19 cells in the 5 mmol·L-1 glucose group,15 mmol·L-1 glucose group,and 30 mmol·L-1 glucose group significantly increased,and those in the 30 mmol·L-1 glucose group were the highest(all P＜0.05);thus,30 mmol·L-1 glucose was selected for subsequent experiments.Compared with the CON group,the KCNQ1OT1 expres-sion,apoptosis rate,cleaved Caspase-3 protein expression,and MDA level in the HG group significantly increased,while the SOD activity and miR-628-5p significantly decreased(all P＜0.05).Compared with the HG+si-NC group,the KC-NQ1OT1 expression,apoptosis rate,cleaved Caspase-3 protein expression,and MDA level in the HG+si-KCNQ1OTl group significantly decreased,while the SOD activity significantly increased(all P＜0.05).Compared with the HG+miR-NC group,miR-628-5p and SOD activity in the HG+miR-628-5p group significantly increased,while the apoptosis rate,cleaved Caspase-3 protein expression,and MDA level significantly decreased(all P＜0.05).Compared with the HG+si-KCNQ1OT1+anti-miR-NC group,miR-628-5p and SOD activity in the HG+si-KCNQ1OT1+anti-miR-628-5p group significantly decreased,while the apoptosis rate,cleaved Caspase-3 protein expression,and MDA level significantly increased(all P＜0.05).Conclusion lncRNA KCNQ1OT1 can reduce the HG-induced apoptosis and oxidative damage of ARPE-19 cells by targeted inhibition of miR-628-5p.
