Hydrogen-rich saline protects immunocytes from radiation-induced apoptosis

BACKGROUND: Radiation often causes depletion of immunocytes in tissues and blood, which results in immunosuppression. Molecular hydrogen (H(2)) has been shown in recent studies to have potential as a safe and effective radioprotective agent through scavenging free radicals. This study was designed to test the hypothesis that H(2) could protect immunocytes from ionizing radiation (IR). MATERIAL/METHODS: H(2) was dissolved in physiological saline or medium using an apparatus produced by our department. A 2-[6-(4′-hydroxy) phenoxy-3H-xanthen-3-on-9-yl] benzoate (HPF) probe was used to detect intracellular hydroxyl radicals (•OH). Cell apoptosis was evaluated by annexin V-FITC and Propidium iodide (PI) staining as well as the caspase 3 activity. Finally, we examined the hematological changes using an automatic Sysmex XE 2100 hematology analyzer. RESULTS: We demonstrated H(2)-rich medium pretreatment reduced •OH level in AHH-1 cells. We also showed H(2) reduced radiation-induced apoptosis in thymocytes and splenocytes in living mice. Radiation-induced caspase 3 activation was also attenuated by H(2) treatment. Finally, we found that H(2) rescued the radiation-caused depletion of white blood cells (WBC) and platelets (PLT). CONCLUSIONS: This study suggests that H(2) protected the immune system and alleviated the hematological injury induced by IR.