Long non-coding RNA nuclear paraspeckle assembly transcript 1 protects human lens epithelial cells against H2O2 stimuli through the nuclear factor kappa b/p65 and p38/mitogen-activated protein kinase axis

Tianqiu Zhou; Mei Yang; Guowei Zhang; Lihua Kang; Ling Yang; Huaijin Guan

doi:10.21037/atm-20-7365

Long non-coding RNA nuclear paraspeckle assembly transcript 1 protects human lens epithelial cells against H₂O₂ stimuli through the nuclear factor kappa b/p65 and p38/mitogen-activated protein kinase axis

Ann Transl Med. 2020 Dec;8(24):1653. doi: 10.21037/atm-20-7365.

Authors

Tianqiu Zhou¹, Mei Yang¹, Guowei Zhang¹, Lihua Kang¹, Ling Yang¹, Huaijin Guan¹

Affiliation

¹ Eye Institute, Affiliated Hospital of Nantong University, Nantong, China.

Abstract

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 (H₂O₂)-treated human lens epithelial cells (HLECs) and its underlying molecular mechanism.

Methods: HLECs (SRA01/04) were treated with 300 µM H₂O₂ to mimic cataract in vitro. 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 H₂O₂-treated HLECs and age-related cataract (ARC) tissues. Knockdown NEAT1 strongly protected against H₂O₂-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 H₂O₂-induced intracellular ROS production and malondialdehyde (MDA) content, but elevated the glutathione (GSH) activity of H₂O₂-treated cells. Also, it is demonstrated that si-NEAT1 greatly inhibited H₂O₂-induced phosphorylation of NF-кB p65 and p38 MAPK.

Conclusions: This study confirmed that knockdown NEAT1 attenuated H₂O₂-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.

Keywords: Cataract; apoptosis; hydrogen peroxide (H2O2); long non-coding RNA (lncRNA); oxidative stress.