The Impact of Differentiation on Cytotoxicity and Insulin Sensitivity in Streptozotocin Treated SH-SY5Y Cells

Abstract

Recently neuronal insulin resistance was suggested playing a role in Alzheimer’s disease. Streptozotocin (STZ) is commonly used to induce impairment in insulin metabolism. In our previous work on undifferentiated SH-SY5Y cells the compound exerted cytotoxicity without altering insulin sensitivity. Nevertheless, differentiation of the cells to a more mature neuron-like phenotype may considerably affect the significance of insulin signaling and its sensitivity to STZ. We aimed at studying the influence of STZ treatment on insulin signaling in SH-SY5Y cells differentiated by retinoic acid (RA). Cytotoxicity of STZ or low serum (LS) condition and protective effect of insulin were compared in RA differentiated SH-SY5Y cells. The effect of insulin and an incretin analogue, exendin-4 on insulin signaling was also examined by assessing glycogen synthase kinase-3 (GSK-3) phosphorylation. STZ was found less cytotoxic in the differentiated cells compared to our previous results in undifferentiated SH-SY5Y cells. The cytoprotective concentration of insulin was similar in the STZ and LS groups. However, the right-shifted concentration–response curve of insulin induced GSK-3 phosphorylation in STZ-treated differentiated cells is suggestive of the development of insulin resistance that was further confirmed by the insulin potentiating effect of exendin-4. Differentiation reduced the sensitivity of SH-SY5Y cells for the non-specific cytotoxicity of STZ and enhanced the relative significance of development of insulin resistance. The differentiated cells thus serve as a better model for studying the role of insulin signaling in neuronal survival. However, direct cytotoxicity of STZ also contributes to the cell death.

A systematic review for the development of Alzheimer’s disease in in vitro models: a focus on different inducing agents

“…An in-vitro model of Alzheimer’s disease has been developed using streptozotocin in various studies. Streptozotocin can modulate the activity of GSK3 alpha/beta ( Bagaméry et al, 2021 ) and secretase, as observed in both in vitro and in vivo studies. Wang et al found that insulin is able to inhibit betaA4 production by modulating amyloid precursor protein processing via an increase in the activity of alpha secretase and a simultaneous decrease in beta secretase activity in the APP overexpressing mutant SHSY5Y cell line.…”

“…Studies have found that L-glutamate leads to excitotoxicity in human neuroblastoma SH-SY5Y cells when treated with different concentrations (0.1–10 mM) for 12 h. The overactivation of NMDA receptors induces glutamate excitotoxicity due to the large influx of Ca +2 and NO. This causes activation of the JNK signaling pathway, which further activates caspase-3 and pro apoptotic genes, ultimately leading to apoptosis and neuronal cell death ( Bagaméry et al, 2021 ). Previous research has also linked glutamate excitotoxicity to a variety of neurodegenerative diseases, retinal diseases, hyperglycemia, diabetic retinopathy, and type 3 and type 4 diabetes ( Wang et al, 2019 ).…”

Psammaplin A and Its Analogs Attenuate Oxidative Stress in Neuronal Cells through Peroxisome Proliferator-Activated Receptor γ Activation

“…Assays were performed only in undifferentiated SH-SY5Y cells, since this cell line has been widely recognized as a valuable model for oxidative stress. Particularly, undifferentiated cells are more sensitive to oxidative damage and neurotoxins than differentiated neurons, which allows to disclose the neuroprotective potential of compounds against this pathological mechanism. − …”

“…Particularly, undifferentiated cells are more sensitive to oxidative damage and neurotoxins than differentiated neurons, which allows to disclose the neuroprotective potential of compounds against this pathological mechanism. 39 − 42 …”

Anti-amyloidogenic amphipathic arginine-dehydrophenylalanine spheres capped selenium nanoparticles as potent therapeutic moieties for Alzheimer's disease

“…It has been shown that the intra-cerebroventricular (ICV) administration of STZ into the brain of rats leads to memory loss, enhanced oxidative stress, and cell death in AD. 85 Rodents administered with STZ via the ICV mode are a well- established model of dementia. 39 STZ is also utilized to investigate and decipher the cellular mechanisms that may lead to the death of neurons in AD.…”