SOD2 Mediates Amifostine‐Induced Protection against Glutamate in PC12 Cells

Abstract

Background. Cytoprotectant amifostine attenuates radiation-induced oxidative injury by increasing intracellular manganese superoxide dismutase (SOD2) in peripheral tissue. However, whether amifostine could protect neuronal cells against oxidative injury has not been reported. The purpose of this study is to explore the protection of amifostine in PC12 cells. Methods. PC12 cells exposed to glutamate were used to mimic neuronal oxidative injury. SOD assay kit was taken to evaluate intracellular Cu/Zn SOD (SOD1) and SOD2 activities; western blot analysis and immunofluorescence staining were performed to investigate SOD2 protein expression; MTT, lactate dehydrogenase (LDH), release and cell morphology were used to evaluate cell injury degree, and apoptotic rate and cleaved caspase-3 expression were taken to assess apoptosis; mitochondrial superoxide production, intracellular reactive oxygen species (ROS), and glutathione (GSH) and catalase (CAT) levels were evaluated by reagent kits. Results. Amifostine increased SOD2 activity and expression, decreased cell injury and apoptosis, reduced mitochondrial superoxide production and intracellular ROS generation, and restored intracellular GSH and CAT levels in PC12 cells exposed to glutamate. SOD2-siRNA, however, significantly reversed the amifostine-induced cytoprotective and antioxidative actions. Conclusion. SOD2 mediates amifostine-induced protection in PC12 cells exposed to glutamate.

Peroxiredoxin 5 Inhibits Glutamate-Induced Neuronal Cell Death through the Regulation of Calcineurin-Dependent Mitochondrial Dynamics in HT22 Cells

“…In neuronal cells, it can lead to increased ROS and nitric oxide (NO) production, expression of cytotoxic transcription factors, and damaged intracellular Ca 2ϩ homeostasis (excessive Ca 2ϩ influx) (50). Previous studies have addressed the neuroprotective effects of antioxidant enzymes using cell models overexpressing ROS-related enzymes like Cu/Zn superoxide dismutase 1 (SOD1), SOD2, and Prx6 (51)(52)(53). In addition, there are several studies that show that immoderate ROS production is closely related to unstable Ca 2ϩ homeostasis (54).…”

Amifostine Pretreatment Attenuates Myocardial Ischemia/Reperfusion Injury by Inhibiting Apoptosis and Oxidative Stress

“…In vitro, amifostine could protect rat heart myocytes against doxorubicin [28]. In addition, Jia et al found that amifostine can reduce the apoptosis rate of PC12 cells induced by glutamate [29]. In another investigation, WR-1065, the active form of amifostine, can reduce apoptosis of HL-60 cells induced by etoposide [30].…”

Antioxidant and Genoprotective Effects of Amifostine against Irinotecan Toxicity in Human Hepatoma Cells

“…Once dephosphorylated by the membrane-bound alkaline phosphatase (ALP), AMF is activated to a free thiol form (WR-1065), which is preferentially up taken by normal cells, since ALP is more active and efficiently expressed in normal rather than neoplastic tissue [37]. Moreover, in another study found that WR1065, the active free thiol form of amifostine, induces antioxidative ability against radiation via SOD2 in vitro [38,39]. Other studies have been shown the role of SOD2 in amifostine-induced protective effects, SOD2 mediated amifostine-induced antioxidative actions in PC12 cells exposed to glutamate.…”