The Effects of Molecular Hydrogen and Suberoylanilide Hydroxamic Acid on Paraquat-Induced Production of Reactive Oxygen Species and TNF-α in Macrophages.
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 H2for 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. H2and H2combined 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 H2caused 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. H2decreases 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 H2and SAHA.
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Jiaoyang
J
Li
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Xizi
X
Wu
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Yao
Y
Chen
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Renqing
R
Zeng
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Yangzi
Y
Zhao
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Panpan
P
Chang
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Danna
D
Wang
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Qianwen
Q
Zhao
Emergency Department, The First Hospital of China Medical University, Shenyang, China.
Yunlei
Y
Deng
Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
Yongqing
Y
Li
Department of Surgery, University of Michigan, Ann Arbor, MI, USA.
Hasan B
HB
Alam
Emergency Department, The First Hospital of China Medical University, Shenyang, China. chongweixiena@yahoo.com.
Wei
W
Chong
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D006540:Herbicides / Q000494:pharmacology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006859:Hydrogen / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D010269:Paraquat / Q000494:pharmacology*; D000067996:RAW 264.7 Cells; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000187:drug effects; D000077337:Vorinostat
D006540:Herbicides; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D000077337:Vorinostat; D006859:Hydrogen; D010269:Paraquat
SAHA; TNF-α; hydrogen; macrophage; paraquat; reactive oxygen species