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Updated: Apr 27, 2026
Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line
Published on: February 17, 2016
Insulin blocks glutamate-induced neurotoxicity in differentiated SH-SY5Y neuronal cells
Madhavan Nampoothiri1, Neetinkumar D Reddy1, Jessy John1
1Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka 576104, India.
Abstract:
Insulin is a cytokine which promotes cell growth. Recently, a few published reports on insulin in different cell lines support the antiapoptotic effect of insulin. But the reports fail to explain the role of insulin in modulating glutamate-mediated neuronal cell death through excitotoxicity. Thus, we examined the neuroprotective effect of insulin on glutamate-induced toxicity on differentiated SH-SY5Y neuronal cells. Changes in cell viability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) based assay, while apoptotic damage was detected by acridine orange/ethidium bromide and Hoechst staining. Intracellular reactive oxygen species (ROS) accumulation and morphological alterations were also measured. Treatment with glutamate induced apoptosis, elevated ROS levels and caused damage to neurons. Insulin was able to attenuate the glutamate-induced excitotoxic damage to neuronal cells.
Insights
Insulin protects against glutamate-induced excitotoxicity in neuronal cells. This study demonstrates insulin
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Insulin, a cytokine, promotes cell growth and exhibits antiapoptotic effects in various cell lines.
- The role of insulin in modulating glutamate-mediated excitotoxicity in neurons remains unclear.
- Excitotoxicity, driven by excessive glutamate, is a significant factor in neuronal cell death.
Purpose of the Study:
- To investigate the neuroprotective potential of insulin against glutamate-induced excitotoxicity.
- To elucidate insulin's mechanism in mitigating neuronal damage caused by glutamate.
- To assess insulin's impact on cell viability, apoptosis, and reactive oxygen species (ROS) in neuronal cells.
Main Methods:
- Utilized differentiated SH-SY5Y neuronal cells.
- Assessed cell viability using the MTT assay.
- Detected apoptotic damage via acridine orange/ethidium bromide and Hoechst staining.
- Measured intracellular reactive oxygen species (ROS) accumulation and morphological alterations.
Main Results:
- Glutamate treatment induced significant apoptosis, elevated ROS levels, and morphological damage in neurons.
- Insulin treatment significantly attenuated the glutamate-induced excitotoxic damage.
- Insulin effectively reduced apoptosis and ROS accumulation in glutamate-exposed neuronal cells.
Conclusions:
- Insulin demonstrates a significant neuroprotective effect against glutamate-induced excitotoxicity.
- Insulin mitigates neuronal cell death by reducing apoptosis and reactive oxygen species.
- These findings highlight insulin's potential therapeutic role in neurodegenerative conditions involving excitotoxicity.
