| 侯知佑,李静,刘军叶,等.氮氧自由基R-1对人肝细胞L-02的辐射防护作用[J].中华放射医学与防护杂志,2011,31(2):172-176.HOU Zhi-you,LI Jing,LIU Jun-ye,et al.Radioprotective effects of nitroxides R-1 on human liver cellsL-02[J].Chin J Radiol Med Prot,2011,31(2):172-176 |
| 氮氧自由基R-1对人肝细胞L-02的辐射防护作用 |
| Radioprotective effects of nitroxides R-1 on human liver cellsL-02 |
| 投稿时间：2010-09-06 |
| DOI：10.3760/cma.j.issn.0254-5098.2011.02.014 |
| 中文关键词: 氮氧自由基 人肝细胞 辐射防护 γ射线 凋亡 |
| 英文关键词:Nitroxides Human liver cells Radioprotection γ-rays Apoptosis |
| 基金项目:国家自然科学基金(60871086) |
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| 中文摘要: |
| 目的 探讨氮氧自由基R-1(简称R-1)对人肝细胞L-02的辐射防护作用。方法 在L-02细胞培养液中,加入终浓度为0、0.125、0.25、0.5、1、2、4、8、16、32 μmol/L的R-1,作用24、48和72 h。用MTT法测定R-1的毒性作用,以筛选合适的R-1浓度。后续实验选择0.125、0.25、0.5、1 μmol/L 4个浓度组检测其防护作用。设终浓度4 mmol/L的细胞保护剂WR2721为阳性对照组。采用 60Co γ射线照射,吸收剂量为0、1、2、4、8 Gy,照射72 h后,进行MTT比色法。L-02细胞分为2组:照射前30 min加药组和照射后立即加药组。终浓度均为0.25 μmol/L,吸收剂量为4 Gy,照射后72 h进行MTT比色实验。照射后10 d,用克隆形成实验检测不同浓度的R-1对L-02细胞活力的影响。选用防护效果最佳的0.25 μmol/L浓度对细胞进行预处理,分别在4 Gy照射后的24、48和72 h,用倒置显微镜观察细胞形态的变化,Hoechst 33258荧光染色法和流式细胞仪检测细胞凋亡。 结果 当R-1浓度低于1 μmol/L时,与0 μmol/L组相比,L-02细胞各时间点的吸光度(A)值无明显变化;而浓度高于2 μmol/L时,与0 μmol/L组相比,其A值随浓度的增高而下降。选用0、0.125、0.25、0.5、1 μmol/L的浓度组,与照射组相比,R-1各浓度组的A值和克隆形成率明显提高,其中0.25 μmol/L组的作用最明显。与照射组相比,0.25 μmol/L预处理组的L-02细胞贴壁好,折光性强,轮廓清晰,凋亡细胞和死亡细胞明显较少。结论 R-1能有效地防护60Co γ射线对L-02细胞的辐射损伤,其防护作用可能与减少细胞凋亡有关。 |
| 英文摘要: |
| 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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