The impact of LncRNA NEAT1 on lens epithelial cell injury under high-glu-cose conditions by modulating the miR-204-5p/TET1 axis
10.13389/j.cnki.rao.2025.0121
- VernacularTitle:LncRNA NEAT1调节miR-204-5p/TET1对高糖条件下晶状体上皮细胞损伤的影响
- Author:
Yi LU
1
;
Fan CHEN
1
;
Huiling YE
1
;
Longqi QIAN
1
Author Information
1. 246000 安徽省安庆市,安庆市立医院
- Publication Type:Journal Article
- Keywords:
LncRNA NEAT1;
miR-204-5p/TET1;
10-11 translocation protein;
high glucose;
lens epithelial cell;
cell injury
- From:
Recent Advances in Ophthalmology
2025;45(9):703-710
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of LncRNA NEAT1-regulated miR-204-5p/ten-eleven translocation protein 1(TET1)on lens epithelial cell(LEC)injury under high-glucose conditions.Methods Target validation was per-formed through RNA pull-down,RNA immunoprecipitation,and dual-luciferase reporter assays.HLEB3 cells were random-ly divided into normal control,high-glucose,si-NEAT1,si-NC+NC-miR-204-5p,and si-NEAT1+miR-204-5p inhibitor groups.Except for the normal control group,all other groups were induced with high glucose.After lentiviral plasmid transfection,RT-qPCR and Western blot were used to detect the expression of LncRNA NEAT1,miR-204-5p,and TET1 in each group of HLEB3 cells.Cell viability and apoptosis were detected by CCK-8 and flow cytometry assays,respectively.Western blot was used to detect the expression of apoptosis-and epithelial-mesenchymal transition(EMT)-related pro-teins.The activity of cellular antioxidant enzymes,reactive oxygen species(ROS),and levels of inflammation-related fac-tors were measured.SD rats were randomly divided into control,diabetic cataract(DC),NEAT1 knockdown,negative control,and NEAT 1 knockdown+miR-204-5p inhibitor groups.DC rat models were induced by intraperitoneal injection of streptozotocin.After lentiviral plasmid intervention,the expression of LncRNA NEAT1,miR-204-5p,and TET1 in rat lens tissues was detected.Lens opacity was scored,and the morphology of rat lens tissues was examined by HE staining.Results LncRNA NEAT1 can target the regulation of miR-204-5p/TET1.Compared with the normal control group,the relative expression of LncRNA NEAT1,TET1 protein,TET1 mRNA,and the proteins Caspase-3,Cleaved Caspase-3,Bax,N-cadherin,MMP9,and MMP2,as well as the levels of ROS,IL-1β,and IL-6,were increased in the high glucose group,while the relative expression of miR-204-5p,cell viability,and the relative expression of Bcl-2 and E-cadherin proteins,and the activities of SOD,CAT,and GSH-Px were decreased(all P＜0.05).Compared with the high glucose group,the above mentioned indicators in HLEB3 cells of the si-NEAT1 group were reversed(all P＜0.05).Compared with the si-NEAT1 group,the relative expression of TET1 protein,TET1 mRNA,and the proteins Caspase-3,Cleaved Caspase-3,Bax,N-cad-herin,MMP9,and MMP2,as well as the levels of ROS,IL-1β,and IL-6,were increased in the si-NEAT1+miR-204-5p in-hibitor group of HLEB3 cells,while the relative expression of miR-204-5p,cell viability,and the relative expression of Bcl-2 and E-cadherin proteins,and the activities of SOD,CAT,and GSH-Px were decreased(all P＜0.05).Compared with the control group,the relative expression of LncRNA NEAT1 and TET1 protein and mRNA,and the lens opacity score were in-creased in the DC group of rats,while the relative expression of miR-204-5p was decreased(all P＜0.05).Compared with the DC group,the above mentioned indicators were reversed in the NEAT1 knockdown group(all P＜0.05).Compared with the NEAT1 knockdown group,the relative expression of TET1 protein and mRNA and the lens opacity score were in-creased in the NEAT1 knockdown+miR-204-5p inhibitor group of rats,while the relative expression of miR-204-5p was de-creased(all P＜0.05).Conclusion Knockdown of LncRNA NEAT1 can alleviate LEC injury under high-glucose condi-tions and mitigate lens opacity and tissue injury in DC rats by up-regulating miR-204-5p to reduce TET1 expression.
