| 林凯丽,徐畅,杜利清,等.Sirt3介导的SOD2在电离辐射中的作用研究[J].中华放射医学与防护杂志,2014,34(8):569-572.Lin Kaili,Xu Chang,Du Liqing,et al.The effect of Sirt3 mediated SOD2 in radiation responses[J].Chin J Radiol Med Prot,2014,34(8):569-572 |
| Sirt3介导的SOD2在电离辐射中的作用研究 |
| The effect of Sirt3 mediated SOD2 in radiation responses |
| 投稿时间：2014-01-02 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.08.003 |
| 中文关键词: 沉默信息调节因子3 超氧化物歧化酶2 活性氧 293T细胞 电离辐射 |
| 英文关键词:Silent information regulator 3 Superoxide dismutase 2 Reactive oxygen species 293 T cells Ionizing radiation |
| 基金项目:卫生部卫生行业科研专项（201002009）；国家自然科学基金（31240052）；天津市自然科学基金（13JCYBJC23500，13JCQNJC11600）；协和青年基金资助和中央高校基本科研业务费专项资金（2012G01） |
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| 全文下载次数: 2012 |
| 中文摘要: |
| 目的 观察293T细胞经不同剂量137Cs γ射线照射后，细胞中沉默信息调节因子3（sirt3）及相关抗氧化因子的变化，研究sirt3基因在辐射防护中的抗氧化活性机理。方法 采用137Cs对293T细胞分别进行2、4、8 和16 Gy照射，采用Real-Time PCR的方法，检测照后6、12、24和48 h，细胞中sirt3和超氧化物歧化酶2（SOD2）mRNA的表达水平变化；采用免疫印迹法，检测细胞中sirt3和SOD2蛋白水平随时间的变化；采用siRNA干扰和抑制剂处理的方法，检测sirt3与SOD2的作用关系；采用SOD Assay Kit-WST试剂盒检测细胞中SOD2酶活力的变化；流式细胞术检测细胞中活性氧（ROS）随时间的变化。结果 各组细胞中sirt3和SOD2的mRNA水平和蛋白质水平均表现出不同程度的上升，与0 Gy组比较，分别在12 h（t=6.75、13.59、6.59、10.13，P<0.05）和24 h（t=6.80、8.73、11.09，P<0.05）达到最大值；sirt3 siRNA干扰和抑制剂处理结果显示sirt3是SOD2的上游调控因子；照射24 h后细胞中SOD2的活性显著增加，与0 Gy组比较，差异有统计学意义（t=46.04、23.19、26.28、14.70，P<0.05），细胞中ROS水平也发生相应变化。结论 sirt3可能通过对细胞中SOD2的表达量及活性的调控加强抗氧化保护系统，为临床辐射防护和治疗提供实验依据。 |
| 英文摘要: |
| Objective To study the effects of the silent information regulator 3 (sirt3) and its relative anti-oxidation factors on ionizing radiation (IR)-induced damage in 293T cells. Methods 293T cells irradiated with 2, 4, 8, 16 Gy of γ -rays, after 6, 12, 24 and 48 h of exposure, the expressions of sirt3 mRNA and SOD2 protein were examined by Real-Time PCR and immunoblotting assay, respectively. sirt3 siRNA was applied to knock down sirt3 in the cells. SOD2 activities were measured by a WST kit and the cellular ROS level was detected by a flow cytometry. Results The mRNA levels of sirt3 and SOD2 in each group campared with the 0 Gy group were reached the maximum at 12 h(t=6.75, 13.59, 6.59, 10.13,P<0.05)and 24 h(t=6.80, 8.73, 11.09,P<0.05), and protein levels also increased. It was found that sirt3 was a upstream regulatory factor of SOD2. The activity of SOD2 were much higher at 24 h(t=46.04, 23.19, 26.28, 14.70,P<0.05)than that in the 0 Gy group as well as the level of ROS. Conclusion Sirt3 may strengthen antioxidant protection system through regulation of the SOD2 expression and activity, and possibly plays an important role in IR-induced damage response and clinic treatment. |
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