Antiproliferative Effect of Elastin-Derived Peptide VGVAPG on SH-SY5Y Neuroblastoma Cells

Abstract

Throughout the lifetime of humans, the amount of stem cells and the rate of cell proliferation continue to decrease. Reactive oxygen species (ROS) are one among the many factors that promote stem cell aging. Both a decrease in the level of stem cells and increase in ROS production can lead to the development of different neurodegenerative diseases. This study was conducted to determine how the VGVAPG peptide, liberated from elastin during the aging process and under pathological conditions, affects ROS production and activities of antioxidant enzymes in undifferentiated, proliferating SH-SY5Y cells. SH-SY5Y cells were maintained in Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 supplemented with 10% heat-inactivated fetal bovine serum (FBS). After treating the SH-SY5Y cells with VGVAPG peptide, we measured ROS production; cell metabolism, proliferation, and expression; and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). We demonstrated that the VGVAPG peptide increases GPx expression and activity, whereas it decreases CAT expression in SH-SY5Y cells. Silencing of the GLB1 gene prevents changes in GPx activity. Despite the fact that the VGVAPG peptide increases GPx expression, it increases the ROS level. Moreover, the VGVAPG peptide decreases SH-SY5Y proliferation, which is prevented by the ROS scavenger N-acetyl-L-cysteine. Our data suggest that ROS production and decreased proliferation of SH-SY5Y cells are the results of excitotoxicity meditated through close unrecognized molecular pathways. More research is needed to elucidate the unknown mechanism of action of VGVAPG peptide in the nervous system.

Specific role of N-methyl-D-aspartate (NMDA) receptor in elastin-derived VGVAPG peptide-dependent calcium homeostasis in mouse cortical astrocytes in vitro

“…Our previous studies have shown that the VGVAPG peptide increases ROS production in an EBP-dependent manner in mouse primary astrocytes in vitro 32 . Similarly, an increase in ROS production after EDP stimulation has been reported in monocytes, human fibroblasts, and a neuroblastoma (SH-SY5Y) cell line 29,30,33 . Moreover, EDPs have been described to enhance the activities of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), or glutathione peroxidase (GPx), and increase lipid peroxidation in human fibroblasts 40 .…”

“…Moreover, EDPs have been described to enhance the activities of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), or glutathione peroxidase (GPx), and increase lipid peroxidation in human fibroblasts 40 . Furthermore, the VGVAPG peptide increases the expression and activity of GPx in SH-SY5Y cells 33 . On the other hand, the VGVAPG peptide reduced ROS production in neutrophils in controls and patients with stable chronic obstructive pulmonary disease 28 .…”

“…However, such data suggest that the effects of EDPs are cell and/or tissue dependent. In a previous study, we found that the VGVAPG peptide increases ROS production in an EBP-dependent manner both in mouse astrocytes in vitro 32 and a human neuroblastoma (SH-SY5Y) cell line 33 . It is well known that calcium influx the NMDAR stimulates ROS production 34 .…”

Elastin-derived peptide VGVAPG decreases differentiation of mouse embryo fibroblast (3T3-L1) cells into adipocytes

“…Because resistin is strictly controlled by PPARγ activation, our data allow to assume that VGVAPG peptide by interaction with this receptor can control resistin gene expression [88]. Interaction between PPARγ and VGVAPG has been proven in our previous study, in which VGVAPG peptide was shown to affect PPARγ expression and PPARγ-mediated effects in mouse astrocytes and SH-SY5Y cells [31,49]. As mentioned above, PPARγ is involved in resistin expression, which is strictly related to obesity and type 2 diabetes.…”

“…It is well known that EDPs or VGVAPG increase ROS production in different cells such as murine monocytes and astrocytes, human fibroblasts, and neuroblastoma (SH-SY5Y) cells [49][50][51][52][53]. Moreover, ROS are key signalling molecules that play an important role in the progression of inflammatory disorders.…”