UNASSIGNED: EA.hy926 cells were divided into seven groups: a control group; 1, 2, or 4 Gy radiation; and 10 μg/mL BG pretreatment for 24 h before 1, 2, or 4 Gy irradiation. Cell survival was assessed by colony formation assay. Cell cycles, apoptosis, DNA damage, and reactive oxygen species (ROS) levels were measured through flow cytometry. The level of malondialdehyde and antioxidant enzyme activities were analyzed using assay kits. The activation of NF-κB was analyzed using western blotting and a transcription factor assay kit. The expression of downstream target genes was detected by western blotting.
UNASSIGNED: BG pretreatment significantly increased the survival of irradiated cells, improved cell cycle progression, and decreased DNA damage and apoptosis. The levels of ROS and malondialdehyde were also decreased by BG. Further study indicated that BG increased the antioxidant enzyme activities, activated Src, and promoted NF-κB activation, especially for the p65, p50, and RelB subunits. The activated NF-κB upregulated the expression of antioxidant protein MnSOD, DNA damage-response and repair-related proteins BRCA2 and Hsp90α, and antiapoptotic protein Bcl-2.
UNASSIGNED: Our results demonstrated that BG protects EA.hy926 cells from high linear-energy-transfer carbon-ion irradiation damage through the upregulation of prosurvival signaling triggered by the interaction of BG with its receptor. This confirms that BG is a promising radioprotective agent for heavy-ion exposure.
■EA。将hy926细胞分为七个组:对照组;1、2或4Gy辐射;在1、2或4Gy辐射之前,10μg/mLBG预处理24小时。通过集落形成测定评估细胞存活。细胞周期,凋亡,DNA损伤,通过流式细胞术测量活性氧(ROS)水平。使用测定试剂盒分析丙二醛水平和抗氧化酶活性。使用蛋白质印迹和转录因子测定试剂盒分析NF-κB的活化。通过蛋白质印迹法检测下游靶基因的表达。
■BG预处理显著增加了照射细胞的存活率,改善细胞周期进程,减少DNA损伤和细胞凋亡。BG也降低了ROS和丙二醛的水平。进一步的研究表明,BG增加了抗氧化酶的活性,激活的Src,并促进NF-κB的激活,特别是p65、p50和RelB亚基。活化的NF-κB上调抗氧化蛋白MnSOD的表达,DNA损伤反应和修复相关蛋白BRCA2和Hsp90α,和抗凋亡蛋白Bcl-2。
■我们的结果表明BG可以保护EA。hy926细胞通过BG与其受体相互作用触发的前生存信号上调,从而受到高线性能量转移碳离子辐照损伤。这证实BG是用于重离子暴露的有前途的辐射防护剂。