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Purpose: Hydroxyl radicals play an important role in ionizing radiation-induced cellular damage which has been demonstrated previously.Since most of the ionizing radiation-induced cellular damage is caused by hydroxyl radicals, this study was designed to test the hypothesis that hydrogen-rich PBS (PBS saturated with molecular hydrogen) which is easy to use can be an effective radioprotective agent in vitro.Materials and Methods:Colony-forming assay was performed in our work.Calculated numbers of cells were plated to enable normalization for plating efficiencies.Pretreated cells were then irradiated with 0, 2, 4, 6 or 8 Gy.After incubating for 7 days,plates were fixed with 70% EtOH and stained with 1% methylene blue.Apoptosis was determined by Annexin V-APC and propidium iodide staining using Apoptosis Detection Kit.Endogenous antioxidant was also detected 8-hydroxydeoxyguanosine concentrations of Human Lymphocyte AHH-1 cells were also detected by Commercial Detection Kit and Elisa Kit.Results: It was demonstrated that treating cells with hydrogen-rich PBS before irradiation could significantly inhibit IR-induced apoptosis, increase viability of Human intestinal crypt HIEC cells and could significantly increase endogenous antioxidant, decrease malondialdehyde and 8-hydroxydeoxyguanosine concentrations of Human Lymphocyte AHH-1 cells.Conclusion : We suggest that hydrogen has a potential as an effective and safe radioprotective agent which give us more hope for the protection from irradiation.