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Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway

Sodium azide (NaN(3)), an inhibitor of cytochrome oxidase, induces the release of excitotoxins via an energy impairment and this, in turn, results in neurodegeneration. The present study aimed to investigate the toxic effects NaN(3) on apoptosis of PC12 cells and its mechanism of action in peroxisom...

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Autores principales: Zuo, Yuanyi, Hu, Jun, Xu, Xuehua, Gao, Xiangting, Wang, Yun, Zhu, Shaohua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6390015/
https://www.ncbi.nlm.nih.gov/pubmed/30664159
http://dx.doi.org/10.3892/mmr.2019.9853
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author Zuo, Yuanyi
Hu, Jun
Xu, Xuehua
Gao, Xiangting
Wang, Yun
Zhu, Shaohua
author_facet Zuo, Yuanyi
Hu, Jun
Xu, Xuehua
Gao, Xiangting
Wang, Yun
Zhu, Shaohua
author_sort Zuo, Yuanyi
collection PubMed
description Sodium azide (NaN(3)), an inhibitor of cytochrome oxidase, induces the release of excitotoxins via an energy impairment and this, in turn, results in neurodegeneration. The present study aimed to investigate the toxic effects NaN(3) on apoptosis of PC12 cells and its mechanism of action in peroxisome proliferator-activated receptor γ co-activator 1-α (Pgc-1α)-associated signaling pathways. To induce apoptosis, PC12 cells were exposed to NaN(3) (0, 5, 10, 20, 40 and 80 mM) for 12, 24, 48 and 72 h. Cell viability was determined by CCK-8 assay. DAPI staining was employed to additionally examine apoptotic cells and their nuclear changes. Production of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm) and apoptotic rate were also assessed by flow cytometry. Cellular ATP content was estimated by firefly luciferase assay. In addition, the expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), phosphorylated (p)-Ca(2+)/calmodulin-dependent protein kinase (CaMK), p-p38 mitogen-activated protein kinase (p38 MAPK), Pgc-1α, nuclear respiratory factor (Nrf)-1, mitochondrial transcription factor A (Tfam), p-extracellular signal-regulated kinase (Erk)1/2, Nrf-2 and complex IV (Cox IV) were determined by western blot analysis. The data suggested that NaN(3) may induce PC12 cell injury and dose-dependently decrease the cell viability. The expression levels of pro-apoptotic proteins Bax and cytochrome c were upregulated, while the expression levels of anti-apoptotic proteins procaspase-3 and Bcl-2 were downregulated. In addition, the phosphorylation of MAPK and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) family members including pan-calcineurin A was increased, in particular the ratios of p-p38/p38 and p-CaMKII/CaMKII. However, the expression levels of Pgc-1α and its associated proteins, including Nrf-1/2, Tfam and p-Erk1/2 were decreased. In addition, mitochondria were the target organelles of NaN(3)-induced toxicity in PC12 cells, which moderated the dissipation of ΔΨm, preserved the cellular ATP content, promoted the production of ROS and increased the apoptotic rate. These results suggested that NaN(3) induced cell death in PC12 cells via Pgc-1α-associated signaling pathways and provided a theoretical basis for additional investigation of the neurotoxic mechanism of NaN(3), with applications in neurodegenerative disorders.
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spelling pubmed-63900152019-03-07 Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway Zuo, Yuanyi Hu, Jun Xu, Xuehua Gao, Xiangting Wang, Yun Zhu, Shaohua Mol Med Rep Articles Sodium azide (NaN(3)), an inhibitor of cytochrome oxidase, induces the release of excitotoxins via an energy impairment and this, in turn, results in neurodegeneration. The present study aimed to investigate the toxic effects NaN(3) on apoptosis of PC12 cells and its mechanism of action in peroxisome proliferator-activated receptor γ co-activator 1-α (Pgc-1α)-associated signaling pathways. To induce apoptosis, PC12 cells were exposed to NaN(3) (0, 5, 10, 20, 40 and 80 mM) for 12, 24, 48 and 72 h. Cell viability was determined by CCK-8 assay. DAPI staining was employed to additionally examine apoptotic cells and their nuclear changes. Production of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm) and apoptotic rate were also assessed by flow cytometry. Cellular ATP content was estimated by firefly luciferase assay. In addition, the expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), phosphorylated (p)-Ca(2+)/calmodulin-dependent protein kinase (CaMK), p-p38 mitogen-activated protein kinase (p38 MAPK), Pgc-1α, nuclear respiratory factor (Nrf)-1, mitochondrial transcription factor A (Tfam), p-extracellular signal-regulated kinase (Erk)1/2, Nrf-2 and complex IV (Cox IV) were determined by western blot analysis. The data suggested that NaN(3) may induce PC12 cell injury and dose-dependently decrease the cell viability. The expression levels of pro-apoptotic proteins Bax and cytochrome c were upregulated, while the expression levels of anti-apoptotic proteins procaspase-3 and Bcl-2 were downregulated. In addition, the phosphorylation of MAPK and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) family members including pan-calcineurin A was increased, in particular the ratios of p-p38/p38 and p-CaMKII/CaMKII. However, the expression levels of Pgc-1α and its associated proteins, including Nrf-1/2, Tfam and p-Erk1/2 were decreased. In addition, mitochondria were the target organelles of NaN(3)-induced toxicity in PC12 cells, which moderated the dissipation of ΔΨm, preserved the cellular ATP content, promoted the production of ROS and increased the apoptotic rate. These results suggested that NaN(3) induced cell death in PC12 cells via Pgc-1α-associated signaling pathways and provided a theoretical basis for additional investigation of the neurotoxic mechanism of NaN(3), with applications in neurodegenerative disorders. D.A. Spandidos 2019-03 2019-01-14 /pmc/articles/PMC6390015/ /pubmed/30664159 http://dx.doi.org/10.3892/mmr.2019.9853 Text en Copyright: © Zuo et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Zuo, Yuanyi
Hu, Jun
Xu, Xuehua
Gao, Xiangting
Wang, Yun
Zhu, Shaohua
Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title_full Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title_fullStr Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title_full_unstemmed Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title_short Sodium azide induces mitochondria-mediated apoptosis in PC12 cells through Pgc-1α-associated signaling pathway
title_sort sodium azide induces mitochondria-mediated apoptosis in pc12 cells through pgc-1α-associated signaling pathway
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6390015/
https://www.ncbi.nlm.nih.gov/pubmed/30664159
http://dx.doi.org/10.3892/mmr.2019.9853
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