| HOU Zhi-you,LI Jing,LIU Jun-ye,et al.Radioprotective effects of nitroxides R-1 on human liver cellsL-02[J].Chinese Journal of Radiological Medicine and Protection,2011,31(2):172-176 |
| Radioprotective effects of nitroxides R-1 on human liver cellsL-02 |
| Received:September 06, 2010 |
| DOI：10.3760/cma.j.issn.0254-5098.2011.02.014 |
| KeyWords:Nitroxides Human liver cells Radioprotection γ-rays Apoptosis |
| FundProject:国家自然科学基金(60871086) |
| Author Name | Affiliation | | HOU Zhi-you | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | LI Jing | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | LIU Jun-ye | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | ZHOU Yong-chun | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | LIU Hai-qiang | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | ZHANG Yan-jun | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | SUN Xiao-li | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China | | GUO Guo-zhen | Department of Radiation Medicine, the Faculty of Preventive Medicine, the Fourth Military Medical University, Xian 710032, China |
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| Abstract:: |
| Objective To investigate the protective effects of the nitroxides R-1 on human liver cells exposed to ionizing radiation. Methods Human liver cells L-02 were cultured and irradiated with 60Co γ-rays at the doses of 0, 1,2, 4, and 8 Gy, in order to screen the proper irradiation dose. WR2721 at the terminal concentration of 4 mmol/L was used as positive control. L-02 cells irradiated with 4 Gy were added with R-1 at the terminal concentration of 0.25 μmol/L at 30 min before irradiation or immediately after irradiation. MTT method was used to screen the proper conditions for follow-up experiment 72 h later. L-02 cell culture fluid was added with R-1 at the concentrations of 0, 0.125, 0.25, 0.5, and 1 μmol/L,respectively for 30 min before irradiation at the doses of 0, 1, 2, 4, and 8 Gy to calculate clone formation rate at 10 d post-irradiation. L-02 cells were cultured and divided into 4 groups: control group without any treatment, drug group pretreated by 0.25 μmol/L R-1 only, irradiation group, irradiated at 4 Gy only, and drug+irradiation group with combination of 0.25 μmol/L R-01 and 4 Gy irradiation. The inverted microscopy and Hoechst 33258 staining and flow cytometry were used to observe the apoptosis of the cells at 24, 48, and 72 h later. Results Nitroxides R-1 did not inhibit the viability of L-02 cell when its concentration was less than 1 μmol/L and it inhibited the L-02 cell growth when the concentration was higher than 2 μmol/L. The A value and colony formation rate of different concentration of R-1 groups were all higher than those of the irradiation group, and the effect of the 0.25 μmol/L drug concentration group was the most significant. Consequently, the concentration 0.25 μmol/L was selected for follow-up experiment. Compared with the irradiation group, the L-02 cells of the pretreatment group showed solid adherence, increased refraction, clear outline, less apoptotic and dead cells at 4 Gy post-irradiation. Conclusions Nitroxides R-1 can protect the human liver cells from 60Co γ-ray induced injury effectively. The mechanism of its protective effect may be the reduction of apoptosis. |
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