4.5 Article

Effects of Manganese and Iron, Alone or in Combination, on Apoptosis in BV2 Cells

Journal

BIOLOGICAL TRACE ELEMENT RESEARCH
Volume -, Issue -, Pages -

Publisher

SPRINGERNATURE
DOI: 10.1007/s12011-023-03792-z

Keywords

Manganese; Iron; Neurotoxicity; Apoptosis; ROS; Antagonistic; Combined exposure

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The aim of this study was to investigate the effects of manganese and iron on apoptosis of BV2 microglia cells, both individually and in combination. The results showed that exposure to manganese, iron, or their combination induced cell activation, increased ROS production, inflammatory cytokine expression, and apoptosis. Additionally, iron was found to reduce the toxicity of manganese, and there was an antagonistic effect between manganese and iron.
The aim of study was to address the effects of manganese and iron, alone and in combination, on apoptosis of BV2 microglia cells, and to determine if combined exposure to these metals augments their individual toxicity. We used a murine microglial BV2 cell line. Cell cytotoxicity was analyzed by propidium iodide (PI) exclusion assay. Cell ROS production was analyzed by 2 & PRIME;, 7 & PRIME;-dichlorofluorescin diacetate (DCFH-DA) probe staining. Pro-inflammatory cytokine production was monitored by ELISA. Cell apoptosis was analyzed by PE Annexin V/7-AAD staining. Mitochondrial membrane integrity was analyzed by flow cytometry. We used immunoblotting to analyze the effect of manganese, iron alone, or their combined exposure on the activation of caspase9, P53, Bax, and Bcl2 apoptosis signaling pathways. Caspase3 activity was determined using a Colorimetric. Manganese, iron, and their combined exposure for 24 h induced the activation of BV2 microglia cells and increased ROS production and the expression of the inflammatory cytokines, IL-1 & beta; and TNF-& alpha;. And we also found that the apoptosis rate increased, mitochondrial membrane potential decreased, apoptosis-related proteins caspase9, P53, Bax, and Bcl2 expression increased, and caspase3 activity increased. Furthermore, we found that combined manganese-iron cytotoxicity was lower than that induced by manganese exposure alone. Manganese, iron alone, or their combination exposure can induce apoptosis in glial cells. Iron can reduce the toxicity of manganese, and there is an antagonistic effect between manganese and iron.

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