Related Experiment Video
Updated: Feb 27, 2026
Measurements of Physiological Stress Responses in C. Elegans
Published on: May 21, 2020
MicroRNA-144 modulates oxidative stress tolerance in SH-SY5Y cells by regulating nuclear factor erythroid 2-related
Chunlei Zhou1, Lan Zhao2, Jian Zheng1
1Medical Laboratory of Tianjin First Center Hospital, Tianjin 300192, China.
Abstract:
Genome-wide analysis of miRNA expression has revealed increased levels of miR-144 in the brains of Alzheimer's disease (AD) patients. Nuclear factor erythroid 2-related factor 2 (NRF2), a potential target of miR-144, is a central regulator of antioxidant response, and plays an important role in glutathione (GSH) biosynthesis and recycling. In this study, miR-144 mimic was used to over express miR-144. Aβ (1-42) was used to induce oxidative stress in SH-SY5Y cells. Cell viability and intracellular reactive oxygen species (ROS) were assessed to identify the effects of miR-144 on oxidative stress status. GSH and glutathion peroxidase (GPX) activities were detected to reveal the effect of miR-144 on GSH accumulation. To understand the effects of miR-144 on GSH biosynthesis and recycling, intracellular GPX1, glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), glutathione reductase (GR), and NRF2 expression were detected by western blot and Real-time PCR. In oxidative stress conditions, miR-144 increased the intracellular accumulation of ROS, reduced cell viability, reduced the activities of GSH and antioxidant enzymes, GPX1, and decreased the expression of GCLC, GCLM, GR and NRF2. In conclusion, miR-144 modulates oxidative stress tolerance by regulating NRF2 expression and GSH generation, which may contribute to the pathogenesis of AD.
Insights
Elevated miR-144 in Alzheimer's disease brains worsens oxidative stress. This microRNA reduces cell viability and antioxidant defenses by targeting NRF2, potentially contributing to disease progression.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Alzheimer's disease (AD) is linked to oxidative stress.
- MicroRNAs (miRNAs) regulate gene expression and are implicated in AD pathogenesis.
- Nuclear factor erythroid 2-related factor 2 (NRF2) is a key regulator of cellular antioxidant responses.
Purpose of the Study:
- To investigate the role of miR-144 in regulating oxidative stress in Alzheimer's disease.
- To determine if miR-144 affects the NRF2 pathway and glutathione (GSH) metabolism.
Main Methods:
- Overexpression of miR-144 using a miR-144 mimic in SH-SY5Y cells.
- Induction of oxidative stress using amyloid-beta (Aβ) (1-42).
- Assessment of cell viability, reactive oxygen species (ROS), GSH levels, antioxidant enzyme activities (GPX), and expression of key proteins (NRF2, GCLC, GCLM, GR) via Western blot and Real-time PCR.
Main Results:
- miR-144 overexpression exacerbated oxidative stress, increasing ROS and decreasing cell viability.
- miR-144 reduced GSH levels and the activity of antioxidant enzymes like GPX1.
- Expression of NRF2, GCLC, GCLM, and GR was downregulated by miR-144 under oxidative stress conditions.
Conclusions:
- miR-144 negatively modulates cellular tolerance to oxidative stress by regulating NRF2 expression and GSH generation.
- These findings suggest that miR-144 plays a role in Alzheimer's disease pathogenesis through oxidative stress pathways.
