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The Effects of Molecular Hydrogen and Suberoylanilide Hydroxamic Acid on Paraquat-Induced Production of Reactive Oxygen Species and TNF-α in Macrophages

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Abstract

The aim of this study is to investigate the effects of molecular hydrogen (H2) and suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on paraquat (PQ)-stimulated production of reactive oxygen species (ROS) and tumor necrosis factor alpha (TNF-α) in macrophages. First, the PQ optimal concentration was determined in RAW264.7 macrophage by treating serum-starved cells with PQ at 0, 0.001, 0.01, 0.1, 1, and 10 mM. We evaluated at 1, 2 and 8 h (1) cell viability (by means of trypan blue exclusion method), (2) intracellular ROS levels (with a fluorescent DCFH-DA probe), and (3) TNF-α level in the culture media (determined by enzyme-linked immunosorbent assay, ELISA). Subsequently, mouse RAW267.4 macrophages were treated with PQ in combination with SAHA and/or H2 for 8 h. PQ exerted a significant stimulatory but nontoxic effect on RAW267.4 macrophages at 0.1 mM. This PQ concentration was used in the subsequent experiments. H2 and H2 combined with SAHA evoked a greater reduction in PQ-induced ROS production than SAHA alone, especially at 2 and 8 h. At 1 and 2 h, treatments involving H2 caused a greater decrease in PQ-induced production of TNF-α than the corresponding treatments without H2. However, at 8 h, treatment with SAHA evoked more pronounced effects on TNF-α than treatment without SAHA. H2 decreases PQ-induced ROS production and attenuates early PQ-induced TNF-α production whereas SAHA reduces the late phase of the PQ-induced TNF-α production in macrophages. The effects are enhanced by the combination of H2 and SAHA.

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Acknowledgments

We express our thanks to the Institute of Endocrinology of First Affiliated Hospital of China Medical University for generously providing all the experimental materials used in this study.

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Authors and Affiliations

  1. Emergency Department, The First Hospital of China Medical University, Shenyang, China

    Jiaoyang Li, Xizi Wu, Yao Chen, Renqing Zeng, Yangzi Zhao, Panpan Chang, Danna Wang, Qianwen Zhao, Yunlei Deng & Wei Chong

  2. Department of Surgery, University of Michigan, Ann Arbor, MI, USA

    Yongqing Li & Hasan B. Alam

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  1. Jiaoyang Li
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  2. Xizi Wu
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Correspondence to Wei Chong.

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Conflict of Interest

This work was supported by the Science and Technology Program of Liaoning Province (Grant number 201202289).

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Jiaoyang Li and Xizi Wu contributed equally to this work.

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Li, J., Wu, X., Chen, Y. et al. The Effects of Molecular Hydrogen and Suberoylanilide Hydroxamic Acid on Paraquat-Induced Production of Reactive Oxygen Species and TNF-α in Macrophages. Inflammation 39, 1990–1996 (2016). https://doi.org/10.1007/s10753-016-0434-z

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  • DOI: https://doi.org/10.1007/s10753-016-0434-z

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