Long non-coding RNA nuclear paraspeckle assembly transcript 1 protects human lens epithelial cells against HO stimuli through the nuclear factor kappa b/p65 and p38/mitogen-activated protein kinase axis.

BACKGROUND

Long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) plays a regulatory role in many biological processes; however, its role in cataracts has yet to be illuminated. This study aimed to investigate the protective role of NEAT1 in hydrogen peroxide (HO)-treated human lens epithelial cells (HLECs) and its underlying molecular mechanism.

METHODS

HLECs (SRA01/04) were treated with 300 µM HO to mimic cataract . Cell viability was detected by performing an MTT assay and EdU staining. Flow cytometry was carried out to detect apoptosis of HLECs. DNA damage was examined using γ-H2A histone family member X staining. and reactive oxygen species (ROS) production was measured using 2',7'dichlorofluorescin diacetate staining. The expression levels of lncRNA and proteins were detected with quantitative real-time polymerase chain reaction and western blot, respectively.

RESULTS

The expression of NEAT1 was observed to be increased in HO-treated HLECs and age-related cataract (ARC) tissues. Knockdown NEAT1 strongly protected against HO-induced cell death and also regulated the expression of cleaved caspase-3, B-cell lymphoma 2, and Bcl-2-associated X protein. Further, knockdown NEAT1 also significantly suppressed HO-induced intracellular ROS production and malondialdehyde (MDA) content, but elevated the glutathione (GSH) activity of HO-treated cells. Also, it is demonstrated that si-NEAT1 greatly inhibited HO-induced phosphorylation of NF-кB p65 and p38 MAPK.

CONCLUSIONS

This study confirmed that knockdown NEAT1 attenuated HO-induced damage in HLECs, and inhibited the oxidative stress and apoptosis of HLECs via regulating nuclear factor-kappa B (NF-κB) p65 and p38 MAPK signaling. It may provide a potential target for clinical treatment of cataracts.