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SOD3 Ameliorates H2O2-Induced Oxidative Damage in SH-SY5Y Cells by Inhibiting the Mitochondrial Pathway

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Abstract

This study was designed to investigate the protective effects of extracellular superoxide dismutase (SOD3) against hydrogen peroxide (H2O2) induced damage in human neuroblastoma SH-SY5Y cells and to elucidate the mechanisms responsible for this beneficial effect. SOD3-overexpressing SH-SY5Y cells were generated by adenoviral vector-mediated infection, and H2O2 was then added into the cell culture system to establish an in vitro model of oxidative stress. Cell viability, the generation of intracellular reactive oxygen species (ROS), the expression and activity of antioxidant enzymes, the levels of lipid peroxidation malondialdehyde (MDA), the expression of mitochondrial apoptosis-related genes, and calcium imaging were examined. Following H2O2 exposure, the over-expression of SOD3 promoted the survival of SH-SY5Y cells; decreased the production of ROS, MDA levels, cytochrome C, caspase-3, caspase-9, and Bax gene expression, and calcium levels; and increased the expression and activity of antioxidant enzyme genes and the expression level of Bcl-2. Together, our data demonstrate that SOD3 ameliorates H2O2-induced oxidative damage in neuroblastoma SH-SY5Y cells by inhibiting the mitochondrial pathway and provide new insights into the functional actions of SOD3 on oxidative stress-induced cell damage.

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Acknowledgments

This work was supported mainly by the Chongqing Science and Technology Committee Fund for Cutting-Edge Research Projects (CSTC2014JCYJA10014) and a grant from the Scientific and Technological Research Program of Chongqing Municipal Education Commission (KJ130320). It was also partially supported by grants from the National Natural Science Foundation of China (NSFC Grant Numbers 81001220 and 81370077), and the Fundamental Research Funds for nonprofit public scientific research institutions of Chongqing from Chongqing Science and Technology Commission (Grant no. 2015CSTC-JBKY-01702).

Author Contributions

HY. and H.R.Y. designed this study; R.Y., L.W., Q.Q.F., and H.W. performed the experiments; R.Y. analyzed the data; R.Y. and H.Y. wrote the manuscript.

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    Authors and Affiliations

    1. Research Center of Neuroscience, Chongqing Medical University, Chongqing, 400016, China

      Rong Yang, Qing-Qing Fu & Hua-Rong Yu

    2. Key Laboratory of Birth Defects and Reproductive Health of the National Health and Family Planning Commission, Chongqing Population and Family Planning Science and Technology Research Institute, Chongqing, 400020, China

      Li Wei

    3. Department of Anesthesiology, University Hospital of Hubei University for Nationalities, Enshi, 445000, Hubei, China

      Hua Wang

    4. Affiliated Hospital of the Academy of Military Medical Sciences, Beijing, 100071, China

      Hua You

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    Rong Yang and Li Wei have contributed equally to this work.

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    Yang, R., Wei, L., Fu, QQ. et al. SOD3 Ameliorates H2O2-Induced Oxidative Damage in SH-SY5Y Cells by Inhibiting the Mitochondrial Pathway. Neurochem Res 41, 1818–1830 (2016). https://doi.org/10.1007/s11064-016-1897-x

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