Related Experiment Video
Updated: Jan 9, 2026
Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue
Published on: August 15, 2012
Annexin A5 Protects SH-SY5Y Cells against L-Glutamate-Induced Cytotoxicity
Zahra Abedini1, Marzieh Mehdieh1, Mohammad Ali Takhshid1
1Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Background:
L- glutamate- induced neurotoxicity is linked to neuronal loss in neurodegenerative diseases and stroke. Annexin A5 (ANXA5) is a cytosolic protein that binds calcium in eukaryotic organisms. This study aimed to evaluate the protective effects of a recombinant ANXA5 against L-glutamate-induced cell death and mitochondrial dysfunction in SH-SY5Y cells.
Methods:
ANXA5 was expressed in E. coli and subsequently purified using affinity chromatography. The effect of L-glutamate (0-300 mM) alone or in combination with ANXA5 (0-5µg/mL) on the viability of the SH-SY5Y cells was assessed using the MTT assay. Mitochondrial membrane potential (MMP) dissipation was detected by rhodamine 123 staining and the flow cytometry method. The expressions of Bax, Bcl-2, and NF-E2-Related Factor 2 (Nrf-2) genes were determined by real-time polymerase chain reaction. GraphPad Prism 8.0 was used to analyze the data using either one-way ANOVA or the Kruskal-Wallis test. P<0.05 was considered statistically significant.
Results:
The findings revealed that L-glutamate reduced the cell viability of SH-SY5Y cells in a dose-dependent manner (P<0.001) (IC50=165 mM). Moreover, treating SH-SY5Y cells with 165 mM of L-glutamate increased MMP dissipation, enhanced Bax expression, and reduced the expression of Bcl-2 and Nrf-2, compared to the control group. ANXA5 alone had no significant effects. However, it reversed the effects of L-glutamate on cell death, MMP dissipation, and gene expression in the SH-SY5Y cells.
Conclusion:
The data suggest that ANXA5 can protect SH-SY5Y cells against glutamate-induced cell death and mitochondrial dysfunction, indicating its possible protective effect against glutamate-induced neurodegeneration.
Insights
Annexin A5 (ANXA5) protects against L-glutamate-induced neurotoxicity by preserving cell viability and mitochondrial function in SH-SY5Y cells. This suggests ANXA5 may be a potential therapeutic agent for neurodegenerative diseases.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- L-glutamate neurotoxicity contributes to neuronal loss in conditions like stroke and neurodegenerative diseases.
- Annexin A5 (ANXA5) is a calcium-binding protein with potential cytoprotective roles.
- SH-SY5Y cells are a widely used human neuroblastoma cell line for studying neuronal processes.
Purpose of the Study:
- To investigate the protective effects of recombinant Annexin A5 (ANXA5) against L-glutamate-induced neurotoxicity.
- To assess ANXA5's impact on cell death and mitochondrial dysfunction in a cellular model.
Main Methods:
- Recombinant ANXA5 was expressed and purified.
- SH-SY5Y cell viability was measured using the MTT assay after L-glutamate exposure, with and without ANXA5.
- Mitochondrial membrane potential (MMP) dissipation was analyzed via flow cytometry.
- Gene expression of Bax, Bcl-2, and Nrf-2 was quantified using real-time PCR.
Main Results:
- L-glutamate significantly reduced SH-SY5Y cell viability in a dose-dependent manner (IC50 = 165 mM).
- L-glutamate exposure led to increased MMP dissipation, elevated Bax expression, and decreased Bcl-2 and Nrf-2 expression.
- ANXA5 treatment reversed the detrimental effects of L-glutamate on cell viability, MMP, and gene expression.
Conclusions:
- Annexin A5 demonstrates significant protective effects against L-glutamate-induced cell death and mitochondrial dysfunction in SH-SY5Y cells.
- These findings suggest ANXA5 holds potential as a therapeutic agent to mitigate glutamate-induced neurodegeneration.
More Related Videos
06:54Quantifying Spontaneous Ca2+ Fluxes and their Downstream Effects in Primary Mouse Midbrain Neurons
Published on: September 9, 2020
09:16Exogenous Administration of Microsomes-associated Alpha-synuclein Aggregates to Primary Neurons As a Powerful Cell Model of Fibrils Formation
Published on: June 26, 2018