Early oxidative stress, DNA damage and inflammation resulting from subcutaneous injection of sulfur mustard into mice.

ABSTRACT

Oxidative stress, DNA damage repair, and inflammation are three important reactions of sulfur mustard (SM) exposure. But molecular related chronological events in the earlier stage of SM exposure model are still unclear. In the research, reactive oxygen species (ROS) was measured by using flow cytometry. Cytokines were tested in Luminex method. Myeloperoxidase (MPO), inducible nitric oxide synthase (iNOS), 8-hydroxy-2-deoxyguanosine (8-OHdG) and glutathione (GSH) activity or levels in serum were determined by commercially available kits. Western blot was used to determination of phosphorylated histone 2A.X (γ-H2A.X). Results showed that the oxidative stress biomarker of ROS and 8-OHdG were significantly increased early at 0.5h of SM exposure, but GSH level was decreased at 0.5h. Similarly, SM increased γ-H2A.X level early at 2h, which reached to peak at 8h and recovered to normal at 24h. MPO and iNOS activity were also increased early at 2h and 0.5h respectively. However, all selected inflammation biomarkers, including IL-6, TNF-α, IL-1ß, MCP-1, GM-CSF and IL-10 concentrations are all unchangeable in 2h. The results indicated that oxidative stress and DNA damage had happened more quickly than inflammation reaction. These chronological events may be due to uncovered generation of reactive oxygen species, DNA alkylation and oxidative DNA damage. In conclusion, this research showed that both oxidative stress and DNA damage are earlier events than inflammation in sulfur mustard toxic mouse model.