MicroRNA-668-3p regulates oxidative stress and cell damage induced by Aβ1-42 by targeting the OXR1/p53-p21 axis

Shengyu Li; Lishuo Wu; Meigang Ma; Longxiu Yang; Chao Qin

doi:10.21037/atm-22-3598

MicroRNA-668-3p regulates oxidative stress and cell damage induced by Aβ1-42 by targeting the OXR1/p53-p21 axis

Ann Transl Med. 2022 Sep;10(17):928. doi: 10.21037/atm-22-3598.

Authors

Shengyu Li^#^{1

2}, Lishuo Wu^#^{3

4}, Meigang Ma², Longxiu Yang², Chao Qin²

Affiliations

¹ Department of Neurology, Wuming Hospital of Guangxi Medical University, Nanning, China.
² Department of Neurology, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
³ Department of Neurology, The Fifth Affiliated Hospital of Guangxi Medical University, Nanning, China.
⁴ Department of Neurology, The First People's Hospital of Nanning, Nanning, China.

^# Contributed equally.

Abstract

Background: Alzheimer's disease (AD) is the most common type of dementia in old age and has become a serious social and medical problem threatening human health. We aimed to explore the mechanisms underlying AD development by screening for microRNAs (miRNAs) that affect AD progression and examining their role in AD development.

Methods: Hematoxylin-eosin (HE) staining, immunohistochemistry, and immunofluorescence (IF) were used to analyze the characteristics of the hippocampus, neuron cell separation, and related protein expression in mice. We used Gene Expression Omnibus (GEO) data analysis to screen miRNAs and mRNAs that affect AD progression, and quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blot analysis to determine changes in miRNA and mRNA levels before and after amyloid β (Aβ)1-42 induction. In addition, we used luciferase analysis to examine miRNA and mRNA binding and the effect of miRNA/mRNA interaction on neuronal cell proliferation. Apoptosis and reactive oxygen species (ROS) levels were examined using Cell Counting Kit-8 analysis and flow cytometry (FCM), respectively. The enzyme-linked immunosorbent assay was used to analyze changes in neuronal cell-secreted oxidative stress-related protein levels through miRNA/mRNA interaction.

Results: Oxidative stress levels were significantly increased in the AD mouse model. GEO data analysis revealed 67 dysregulated miRNAs, and miR-668-3p was identified as a potential therapeutic target for AD. We found that the AD and Aβ1-42-induced models showed an increase in miR-668-3p and a decrease in oxidation resistance 1 (OXR1) expression. The luciferase analysis results revealed that miR-668-3p may play a role in AD development by targeting OXR1 and promoting intracellular oxidative stress by activating p53-p21 signaling. The final rescue experiment also confirmed that Aβ1-42-induction decreased cell proliferation, increased apoptosis, increased cell cycle arrest, and promoted oxidative stress. Tenovin-1 (TEN) enhanced the effect of Aβ1-42, and the miR-668-3p inhibitor partially alleviated it, although the effect of the miR-668-3p inhibitor was weakened by TEN.

Conclusions: MiR-668-3p negatively regulated OXR1 expression by targeting OXR1, affecting p53-p21 protein signaling, and regulating cell damage and oxidative stress induced by Aβ1-42. Therefore, miR-668-3p may be a potential therapeutic target for AD.

Keywords: Alzheimer’s disease (AD); OXR1; miR-668-3p; p21; p53.