Annexin A5 ameliorates H2O2-induced cytotoxicity in SH-SY5Y cells
Marzieh Mehdieh1, Gholamreza Rafiei Dehbidi2, Mohammad Hossein Morowvat3
1Division of Medical Biotechnology, Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Background:
A complex interplay between oxidative stress, mitochondrial dysfunction, and apoptosis contributes to the pathogenesis of neurodegenerative diseases (ND). Annexin A5 (ANXA5) is a calcium-binding multifunctional protein. This study aimed to evaluate the possible protective effects of ANXA5 protein against H2O2-induced cell death, reactive oxygen species (ROS) accumulation, mitochondrial membrane potential (MMP) dissipation, and expression of Bax, Bcl2, and NRF2 in SH-SY5Y cells.
Methods:
SH-SY5Y cells were treated with different concentrations of H2O2, and cell viability was determined using an MTT assay. MMP dissipation and ROS accumulation were measured using rhodamine 123 and DCF-DA flow cytometry. The DNA fragmentation assay and real-time PCR were conducted to determine the effects of the treatments on genomic DNA damage and gene expression.
Results:
H2O2 reduced the viability of SH-SY5Y cells in a dose-dependent manner (IC50 = 550µM). Moreover, H2O2 at the IC50 concentration induced ROS accumulation, MMP loss, increased Bax expression, and reduced NRF2 expression in the SH-SY5Y cells. ANXA5 reversed the cytotoxic effects of H2O2 on the SH-SY5Y cell viability, ROS accumulation, MMP loss, and gene expression.
Conclusion:
ANXA5 can protect the SH-SY5Y cells against H2O2-induced oxidative stress, apoptosis, and mitochondrial dysfunction.
Insights
Annexin A5 (ANXA5) protein protects SH-SY5Y cells from hydrogen peroxide (H₂O₂)-induced damage. It mitigates oxidative stress, apoptosis, and mitochondrial dysfunction, offering a potential therapeutic strategy for neurodegenerative diseases.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Neurodegenerative diseases involve oxidative stress, mitochondrial dysfunction, and apoptosis.
- Annexin A5 (ANXA5) is a multifunctional calcium-binding protein.
- Investigating ANXA5's role in protecting against H₂O₂-induced cellular damage is crucial.
Purpose of the Study:
- To evaluate the protective effects of ANXA5 against H₂O₂-induced cell death.
- To assess ANXA5's impact on reactive oxygen species (ROS) and mitochondrial membrane potential (MMP).
- To determine ANXA5's influence on the expression of apoptosis-related genes (Bax, Bcl2) and NRF2.
Main Methods:
- SH-SY5Y cells were exposed to varying H₂O₂ concentrations.
- Cell viability was assessed using MTT assay.
- ROS accumulation, MMP dissipation, and gene expression were measured via flow cytometry and real-time PCR.
Main Results:
- H₂O₂ reduced cell viability in a dose-dependent manner (IC₅₀ = 550µM).
- H₂O₂ induced ROS accumulation, MMP loss, increased Bax, and decreased NRF2 expression.
- ANXA5 treatment reversed these H₂O₂-induced detrimental effects.
Conclusions:
- ANXA5 exhibits protective properties against H₂O₂-induced cellular damage.
- ANXA5 mitigates oxidative stress, apoptosis, and mitochondrial dysfunction in neuronal cells.
- ANXA5 demonstrates potential as a therapeutic agent for neurodegenerative conditions.
