Long non-coding RNA nuclear paraspeckle assembly transcript 1 protects human lens epithelial cells against H <sub>2</sub>O <sub>2</sub> stimuli through the nuclear factor kappa b/p65 and p38/mitogen-activated protein kinase axis

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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 (H ₂O ₂)-treated human lens epithelial cells (HLECs) and its underlying molecular mechanism.

Methods

HLECs (SRA01/04) were treated with 300 µM H ₂O ₂ 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 H ₂O ₂-treated HLECs and age-related cataract (ARC) tissues. Knockdown NEAT1 strongly protected against H ₂O ₂-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 H ₂O ₂-induced intracellular ROS production and malondialdehyde (MDA) content, but elevated the glutathione (GSH) activity of H ₂O ₂-treated cells. Also, it is demonstrated that si-NEAT1 greatly inhibited H ₂O ₂-induced phosphorylation of NF-кB p65 and p38 MAPK.

Conclusions

This study confirmed that knockdown NEAT1 attenuated H ₂O ₂-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.

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Most cited references 35

Record: found
Abstract: found
Article: not found

Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

Kenneth Livak, Thomas D. Schmittgen (2001)

The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).

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Record: found
Abstract: found
Article: not found

The Lens Opacities Classification System III. The Longitudinal Study of Cataract Study Group.

Richard I. Bailey, Peter Friend, T Wu … (1993)

To develop the Lens Opacities Classification System III (LOCS III) to overcome the limitations inherent in lens classification using LOCS II. These limitations include unequal intervals between standards, only one standard for color grading, use of integer grading, and wide 95% tolerance limits. The LOCS III contains an expanded set of standards that were selected from the Longitudinal Study of Cataract slide library at the Center for Clinical Cataract Research, Boston, Mass. It consists of six slit-lamp images for grading nuclear color (NC) and nuclear opalescence (NO), five retroillumination images for grading cortical cataract (C), and five retroillumination images for grading posterior subcapsular (P) cataract. Cataract severity is graded on a decimal scale, and the standards have regularly spaced intervals on a decimal scale. The 95% tolerance limits are reduced from 2.0 for each class with LOCS II to 0.7 for nuclear opalescence, 0.7 for nuclear color, 0.5 for cortical cataract, and 1.0 for posterior subcapsular cataract with the LOCS III, with excellent interobserver agreement. The LOCS III is an improved LOCS system for grading slit-lamp and retroillumination images of age-related cataract.

0 comments Cited 202 times – based on 0 reviews      Review now

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Record: found
Abstract: not found
Article: not found

Suicidal tendencies: apoptotic cell death by caspase family proteinases.

Jason Wolf, D. Green (1999)

0 comments Cited 107 times – based on 0 reviews      Review now

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Author and article information

Journal

Journal ID (nlm-ta): Ann Transl Med

Journal ID (iso-abbrev): Ann Transl Med

Journal ID (publisher-id): ATM

Title: Annals of Translational Medicine

Publisher: AME Publishing Company

ISSN (Print): 2305-5839

ISSN (Electronic): 2305-5847

Publication date (Print and electronic): December 2020

Publication date (Print): December 2020

Volume: 8

Issue: 24

Electronic Location Identifier: 1653

Affiliations

[1]Eye Institute, Affiliated Hospital of Nantong University, Nantong, China

Author notes

Contributions: (I) Conception and design: T Zhou, M Yang, H Guan; (II) Administrative support: L Kang, H Guan; (III) Provision of study materials or patients: G Zhang; (IV) Collection and assembly of data: T Zhou, M Yang; (V) Data analysis and interpretation: H Guan, M Yang; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors.

Correspondence to: Huaijin Guan. 20 Xisi Road, Nantong 226000, China. Email: tog12w@ 123456163.com .

Article

Publisher ID: atm-08-24-1653

DOI: 10.21037/atm-20-7365

PMC ID: 7812193

PubMed ID: 33490165

SO-VID: 6e618b8a-4dbf-4ebc-a8c3-55258d97fdf0

License:

Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0.

History

Date received : 16 October 2020

Date accepted : 02 December 2020

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