Manipulation of HSP70-SOD1 Expression Modulates SH-SY5Y Differentiation and Susceptibility to Oxidative Stress-Dependent Cell Damage: Involvement in Oxotremorine-M-Mediated Neuroprotective Effects

Abstract

The differentiation of neural progenitors is a complex process that integrates different signals to drive transcriptional changes, which mediate metabolic, electrophysiological, and morphological cellular specializations. Understanding these adjustments is essential within the framework of stem cell and cancer research and therapy. Human neuroblastoma SH-SY5Y cells, widely used in neurobiology research, can be differentiated into neuronal-like cells through serum deprivation and retinoic acid (RA) supplementation. In our study, we observed that the differentiation process triggers the expression of Heat Shock Protein 70 (HSP70). Notably, inhibition of HSP70 expression by KNK437 causes a dramatic increase in cell death. While undifferentiated SH-SY5Y cells show a dose-dependent decrease in cell survival following exposure to hydrogen peroxide (H2O2), differentiated cells become resistant to H2O2-induced cell death. Interestingly, the differentiation process enhances the expression of SOD1 protein, and inhibition of HSP70 expression counteracts this effect and increases the susceptibility of differentiated cells to H2O2-induced cell death, suggesting that the cascade HSP70-SOD1 is involved in promoting survival against oxidative stress-dependent damage. Treatment of differentiated SH-SY5Y cells with Oxotremorine-M (Oxo), a muscarinic acetylcholine receptor agonist, enhances the expression of HSP70 and SOD1 and counteracts tert–Butyl hydroperoxide-induced cell death and reactive oxygen species (ROS) generation. It is worth noting that co-treatment with KNK437 reduces SOD1 expression and Oxo-induced protection against oxidative stress damage, suggesting the involvement of HSP70/SOD1 signaling in this beneficial effect. In conclusion, our findings demonstrate that manipulation of the HSP70 signal modulates SH-SY5Y differentiation and susceptibility to oxidative stress-dependent cell death and unravels novel mechanisms involved in Oxo neuroprotective functions. Altogether these data provide novel insights into the mechanisms underlying neuronal differentiation and preservation under stress conditions.

The Effects of Caloric Restriction on Inflammatory Targets in the Prostates of Aged Rats

“…We can speculate that the latter finding may be a response signal by which CR protects prostate tissue against oxidative stress, as already reported in [79,80]. Recently, Scordino and colleagues described Hsp70/SOD1 signaling against oxidative stress damage [81]. The heat shock protein 70 participates in maintaining redox homeostasis through the activation of different intracellular signaling pathways, including increasing the expression and activity of SOD1 [82,83].…”

Unveiling Glucosinolate Diversity in Brassica Germplasm and In Silico Analysis for Determining Optimal Antioxidant Potential

“…Conversely, SIN’s lower affinity for CAT indicates a weaker potential to influence this enzyme’s role in hydrogen peroxide detoxification [ 84 ]. Specific affinities and interactions can guide the development of antioxidant remedies, which has been challenging due to the complexity of oxidative stress mechanisms and the limitations of antioxidant therapy [ 85 ]. These affinities are important for understanding the potential influence of these ligands on modulating the activity of antioxidant enzymes and managing oxidative-stress-related conditions [ 86 ].…”

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 …”