The effect of salmon calcitonin against glutamate-induced cytotoxicity in the C6 cell line and the roles the inflammatory and nitric oxide pathways play

Abstract

Recent evidence has shown that salmon calcitonin (sCT) has positive effects on the nervous system.However, its effect and mechanisms on glutamate-induced cytotoxicity are still unclear. The current experiment was designed to examine the effect of sCT on glutamate-induced cytotoxicity in C6 cells, involving the in ammatory and nitric oxide stress pathways. The study used the C6 glioma cell line. Four cell groups were prepared to evaluate the effect of sCT on glutamate-induced cytotoxicity. The control group was without any treatment. Cells in the glutamate group were treated with 10 mM glutamate for 24 h. Cells in the sCT group were treated with various concentrations (3, 6, 12, 25, and 50 µg/mL) of sCT for 24 hours. Cells in the sCT + glutamate group were pre-treated with various concentrations of sCT for 1 hour and then exposed to glutamate for 24 hours. The cell viability was evaluated with an XTT assay. Nuclear factor kappa b (NF-kB), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), neuronal nitric oxide synthase (nNOS), nitric oxide (NO), cyclic guanosine monophosphate (cGMP), caspase-3, and caspase-9 levels in the cells were measured by ELISA kits. Apoptosis was detected by ow cytometry method. sCT at all concentrations signi cantly improved the cell viability in C6 cells after glutamateinduced cytotoxicity (p < 0.001). Moreover, sCT signi cantly reduced the levels of NF-kB (p < 0.001), TNFα, and IL-6 levels (p < 0.001). sCT also decreased nNOS, NO, and cGMP levels (P < 0.001). Furthermore, it decreased the apoptosis rate and increased the live-cell rate in the ow cytometry (P < 0.001). In conclusion, sCT has protective effects on glutamate-induced cytotoxicity in C6 glial cells by inhibiting in ammatory and nitric oxide pathways. sCT could be a useful supportive agent for people with neurodegenerative symptoms.

Protective Effect of Ethosuximide against Glutamate-Induced Cytotoxicity and Oxidative Stress in C6 Glial Cells

“…Excitotoxicity, which is generated by excessive glutamate stimulation and results in excessive calcium ion influx into the cell, is the basic mechanism in the pathogenesis of these disorders. Calcium enters the cell, which causes mitochondrial malfunction and elevates intracellular nitric oxide levels, inducing cell death processes [10].…”

“…Because of its glutamate-induced cytotoxicity, the C6 Glioma (CRL107) cell line was chosen in this work [10]. The American Type Culture Collection provided C6 Glioma cell lines.…”

“…Cells in the glutamate group were treated with glutamate at dose of 10 mM and then incubated for 24 h. According to the literature, the glutamate concentration (10 mM) that causes the death of 50% of glial cells was chosen [11]. ethosuximide was added to cells in the ethosuximide group at different concentrations (5,10,20, and 40 μM) and then the cells were incubated for 24 hours. Ethosuximide was added to cells in the ethosuximide + glutamate group at different concentrations (5,10,20, and 40 μM) for 1 hours and then10 mM glutamate was applied for 24 hours.…”

“…ethosuximide was added to cells in the ethosuximide group at different concentrations (5,10,20, and 40 μM) and then the cells were incubated for 24 hours. Ethosuximide was added to cells in the ethosuximide + glutamate group at different concentrations (5,10,20, and 40 μM) for 1 hours and then10 mM glutamate was applied for 24 hours.…”

Sugammadex Causes C6 Glial Cell Death and Exacerbates Hydrogen Peroxide-induced Oxidative Stress

“…Preparation of cell homogenates was carried out as previously described 19 . The cells in each group were sampled in sterile tubes and then centrifuged for about 10 minutes at 2000 rpm.…”