Department of Physiology, Sivas Cumhuriyet University School of Medicine, 58140, Sivas, Turkey.
Department of Pharmacology, Sivas Cumhuriyet University School of Pharmacy, Sivas, Turkey.
Metab Brain Dis. 2021 Oct;36(7):1985-1993. doi: 10.1007/s11011-021-00793-6. Epub 2021 Aug 9.
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 inflammatory 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 h. Cells in the sCT + glutamate group were pre-treated with various concentrations of sCT for 1 h and then exposed to glutamate for 24 h. 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 flow cytometry method. sCT at all concentrations significantly improved the cell viability in C6 cells after glutamate-induced cytotoxicity (p < 0.001). Moreover, sCT significantly reduced the levels of NF-kB (p < 0.001), TNF-α, 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 flow cytometry (P < 0.001). In conclusion, sCT has protective effects on glutamate-induced cytotoxicity in C6 glial cells by inhibiting inflammatory and nitric oxide pathways. sCT could be a useful supportive agent for people with neurodegenerative symptoms.
最近的证据表明，鲑鱼降钙素（sCT）对神经系统有积极影响。然而，其对谷氨酸诱导的细胞毒性的作用和机制尚不清楚。本实验旨在研究 sCT 对 C6 细胞中谷氨酸诱导的细胞毒性的影响，涉及炎症和一氧化氮应激途径。该研究使用 C6 神经胶质瘤细胞系。制备了 4 个细胞组来评估 sCT 对谷氨酸诱导的细胞毒性的影响。对照组未进行任何处理。谷氨酸组的细胞用 10 mM 谷氨酸处理 24 小时。sCT 组的细胞用不同浓度（3、6、12、25 和 50 µg/mL）的 sCT 处理 24 小时。sCT + 谷氨酸组的细胞先用不同浓度的 sCT 预处理 1 小时，然后暴露于谷氨酸 24 小时。用 XTT 测定法评估细胞活力。用 ELISA 试剂盒测量细胞核因子 kappa B（NF-kB）、肿瘤坏死因子 alpha（TNF-α）、白细胞介素 6（IL-6）、神经元型一氧化氮合酶（nNOS）、一氧化氮（NO）、环鸟苷单磷酸（cGMP）、半胱天冬酶-3 和半胱天冬酶-9 水平。通过流式细胞术方法检测细胞凋亡。sCT 在所有浓度下均显著提高了谷氨酸诱导的细胞毒性后 C6 细胞的细胞活力（p<0.001）。此外，sCT 显著降低了 NF-kB（p<0.001）、TNF-α 和 IL-6 水平（p<0.001）。sCT 还降低了 nNOS、NO 和 cGMP 水平（P<0.001）。此外，它降低了流式细胞术的细胞凋亡率并增加了活细胞率（P<0.001）。总之，sCT 通过抑制炎症和一氧化氮途径对 C6 神经胶质细胞中的谷氨酸诱导的细胞毒性具有保护作用。sCT 可能是治疗神经退行性疾病症状的有用辅助药物。
