Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells

Abstract

Objective To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl 2 )-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. Methods PC12 cells impaired by CoCl 2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 cells exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. Results CoCl 2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. CoCl 2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 μmol/L. Conclusion These results indicated that Eda could protect PC12 cells from CoCl 2 -induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities.

Combined effects of TNF‐α, IL‐1β, and HIF‐1α on MMP‐2 production in ACL fibroblasts under mechanical stretch: An in vitro study

“…27,28 Our preliminary results showed that CoCl 2 was able to induce HIF-1a at 100 and 500 mM concentrations (Supplementary Figure 2). However, higher concentrations (>500 mM) resulted in apoptosis of ACL fibroblasts (Supplementary Figure 3) therefore, 100 and 500 mM concentration of CoCl 2 were used in the experiments.…”

The neurotoxicity of iron, copper and cobalt in Parkinson’s disease through ROS-mediated mechanisms

“…Cobalt chloride (CoCl 2 ) is a well-known hypoxia mimetic agent. These CoCl 2 -elicited biochemical changes include the production of ROS, a loss of mitochondrial membrane potential, activation of hypoxia inducible factor 1a (HIF-1a) and the expression of a number of genes, such as erythropoietin, vascular endothelial growth factor and endothelin-2/vasoactive intestinal contractor (one of the hypoxia-related factors) (Chandel et al 1998;Guillemin and Krasnow 1997;Yang et al 2004;Zou et al 2002;Kotake-Nara and Saida 2007;Chen et al 2009Chen et al , 2010bYang et al 2011b).…”

“…It is reported that one of the mechanisms underlying CoCl 2 -induced neuronal damage is associated with the production of ROS (Zou et al 2002;Chen et al 2009;Jung et al 2008). CoCl 2 could function as an oxidative stress-inducing factor since Co(II) can react with H 2 O 2 via Fenton-like reaction to produce ROS (Wang et al 1993).…”

Interaction between ROS and p38MAPK contributes to chemical hypoxia-induced injuries in PC12 cells

“…Cobalt chloride (CoCl 2 ) is a well-known hypoxia mimetic agent. There are numerous reports that CoCl 2 mimics the hypoxia response in a number of aspects in various cultured cells, including production of reactive oxygen species (ROS), a loss of mitochondrial membrane potential (MMP), activation of hypoxia inducible factor 1α (HIF-1α) and the expression of a number of genes, such as erythropoietin (EPO), vascular endothelial growth factor (VEGF) and endothelin-2 (ET-2)/vasoactive intestinal contractor (one of the hypoxia-related factors) (1)(2)(3)(4)(5)(6)(7)(8). Therefore, PC12 cells (a cell line derived from rat pheochromocytoma) exposed to CoCl 2 have been used to investigate events and mechanisms underlying neuronal cell death under conditions of hypoxia/ischemia (4,6,9).…”

“…Accumulating evidence suggests that one of the mechanisms of CoCl 2 -induced neuronal damage is associated with its production of ROS (4,6,9). Elevated ROS is capable of attacking nucleic acids, proteins and membrane phospholipids, leading to neuronal apoptosis (10)(11)(12).…”