Robello Elizabeth, Galatro Andrea, Puntarulo Susana
Physical Chemistry-PRALIB, School of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.
Neurotoxicology. 2009 May;30(3):430-5. doi: 10.1016/j.neuro.2009.02.008. Epub 2009 Feb 21.
This study was aimed to assess the content of total Fe, Ferritin (Ft) and labile Fe pool (LIP) in developing rat brain exposed in utero to 1 Gy of gamma-irradiation. A significant increase (2.3-fold) in the total Fe content of the fetal rat brain irradiated in utero was observed from 1 to 4h post-irradiation, as compared to the content in non-irradiated brain. Ft was analyzed by immunoblotting. The Ft protein was composed by 20 kDa subunits. According to the analysis of the band density in the Western blot, the Ft content decreased by 77+/-15% 2h after gamma-irradiation, as compared to the values in non-irradiated samples. The effect of gamma-irradiation on the LIP was studied by both electron paramagnetic resonance (EPR) and by a fluorescence technique employing calcein (CA). A reduction on the LIP was detected at 2h post-irradiation, independently of the methodology employed for the assay. Since NO content increased in the same time frame of LIP decreasing, a protective role for NO is suggested in fetal rat brain exposed to gamma-irradiation. The data presented in this work are the first experimental evidence suggesting that, as part of the network of the cellular response to limit irradiation-dependent injury, a complex interaction between Fe and NO could be triggered.
本研究旨在评估子宫内暴露于1 Gy γ射线照射下的发育中大鼠大脑中的总铁、铁蛋白(Ft)和不稳定铁池(LIP)含量。与未照射大脑中的含量相比,在照射后1至4小时观察到子宫内照射的胎鼠大脑总铁含量显著增加(2.3倍)。通过免疫印迹分析Ft。Ft蛋白由20 kDa亚基组成。根据蛋白质印迹条带密度分析,与未照射样品的值相比,γ射线照射后2小时Ft含量下降了77±15%。通过电子顺磁共振(EPR)和使用钙黄绿素(CA)的荧光技术研究了γ射线照射对LIP的影响。照射后2小时检测到LIP降低,与用于测定的方法无关。由于在LIP降低的同一时间范围内NO含量增加,提示NO在暴露于γ射线照射的胎鼠大脑中具有保护作用。本研究提供的数据首次表明,作为限制辐射依赖性损伤的细胞反应网络的一部分,铁和NO之间可能触发复杂的相互作用。