分次低剂量电离辐射在诱导EA.hy926细胞衰老中的作用
Effects of fractionated low-dose ionizing radiation in the induction of EA.hy926 cell senescence
蔡雅诗 1黄伟旭 2张灵钰 3张敏 2李慧娴 2闻昌勇 2何志妮 4邹剑明 2陈慧峰1
作者信息
- 1. 南方医科大学公共卫生学院,广东广州 510515;广东省职业病防治院,广东广州 510300
- 2. 广东省职业病防治院,广东广州 510300
- 3. 广东省职业病防治院,广东广州 510300;广州医科大学公共卫生学院,广东广州 511436
- 4. 南方医科大学公共卫生学院,广东广州 510515
- 折叠
摘要
目的 探讨分次低剂量电离辐射(LDIR)在诱导EA.hy926细胞衰老中的作用机制.方法 将EA.hy926细胞分为0、50 mGy × 4、100 mGy × 4、200 mGy × 4组,采用X射线辐照后分别培养24、48和72 h,检测细胞衰老相关β半乳糖苷酶(SA-β-gal)染色、衰老相关的细胞周期蛋白依赖性激酶抑制剂CDKN1A和CDKN2A基因mRNA水平、活性氧(ROS)、总抗氧化能力(T-AOC)、磷酸化H2A组蛋白变体X(y-H2AX)水平.结果 LDIR辐照4次后,与对照组相比,24、48和72 h,各剂量组细胞核面积增大,细胞边缘模糊,SA-β-gal阳性面积均增加(P＜0.05);辐照4次后24 和 48 h,100 mGy × 4 组和 200 mGy × 4 组的 CDKN1A 基因 mRNA 水平升高(P＜0.05),48 和 72 h,100 mGy × 4 组和200 mGy × 4组CDKN2A基因mRNA水平升高(P＜0.05);辐照4次后24、48和72 h,各剂量组ROS荧光强度升高(P＜0.05);辐照4次后24 h,100 mGy × 4组和200 mGy × 4组T-AOC水平升高(P＜0.05),48 h和72 h,各剂量组T-AOC水平均升高(P＜0.05);辐照4次后24 h,各剂量组γ-H2AX荧光强度升高(P＜0.05),48和72 h,100 mGy × 4组和200 mGy × 4组γ-H2AX荧光强度均升高(P＜0.05).结论 分次LDIR可通过影响细胞氧化-抗氧化及氧化损伤水平诱导EA.hy926细胞衰老,其中100 mGy和200 mGy效应较明显.
Abstract
Objective To investigate the mechanism of fractionated low-dose ionizing radiation(LDIR)in the induction of EA.hy926 cell senescence.Methods EA.hy926 cells were irradiated with X-ray at 0,50,100,and 200 mGy× 4,re-spectively,and cultured for 24,48,and 72 h.Several indicators were measured,including the levels of cellular senescence-associated β-galactosidase(SA-β-gal)staining,mRNA levels of senescence-associated cell cycle protein-dependent kinase inhibitor genes CDKN1A and CDKN2A,reactive oxygen species(ROS),total antioxidant capacity(T-AOC),and phos-phorylated H2A histone family member X(y-H2AX).Results After 4 fractionated LDIR,compared with the control group,the treatment groups showed increased nucleus area,blurred cell edge,and increased SA-β-gal positive area(P＜0.05)at 24,48 and 72 h.After 4 fractionated LDIR,the mRNA level of CDKN1A increased in the 100 and 200 mGy × 4 groups at 24 and 48 h(P＜0.05),and CDKN2A mRNA level increased in the 100 and 200 mGy × 4 groups at 48 and 72 h(P＜0.05).The fluorescence intensity of ROS increased in treatment groups at 24,48,and 72 h after 4 fractionated LDIR(P＜0.05).After 4 fractionated LDIR,the T-AOC level increased in the 100 and 200 mGy × 4 groups at 24 h(P＜0.05),and T-AOC level increased in all treatment groups at 48 and 72 h(P＜0.05).After 4 fractionated LDIR,γ-H2AX fluorescence intensity increased in all treatment groups at 24 h(P＜0.05),and the fluorescence intensity increased in the 100 and 200 mGy × 4 groups at 48 and 72 h(P＜0.05).Conclusion Fractionated LDIR can induce cellular senescence in EA.hy-926 cells by impacting the cellular oxidation-antioxidation and oxidative damage levels,and the effects were relatively evid-ent at 100 and 200 mGy.
关键词
低剂量电离辐射/氧化应激/DNA损伤/细胞衰老Key words
Low-dose ionizing radiation/Oxidative stress/DNA damage/Cellular senescence引用本文复制引用
基金项目
国家自然科学基金(81302387)
广东省基础与应用基础研究基金(2019A1515011969)
广东省基础与应用基础研究基金(2022A1515012421)
广东省基础与应用基础研究基金(2023A1515010414)
广州市科技计划(202002030031)
出版年
2024
NETL
NSTL
万方数据