摘要
目的研究Sod/Gpx/Cat抗氧化酶系列基因在小鼠骨髓抑制和再生过程中的动态表达,了解氧化应激在骨髓再生过程中的作用。方法制备经5-氟尿嘧啶(5-Fu)注射后的骨髓抑制和再生C57BL/6小鼠模型,分别在5-Fu注射前(第0天)和注射后第3、7、11、14天各断颈处死6只小鼠(分别记为0d组、3d组、7d组、11d组、14d组),收集小鼠骨髓单核细胞。Trizol试剂抽提小鼠骨髓单核细胞总RNA,采用Affymetrix小鼠全基因组cDNA 430A芯片检测小鼠骨髓细胞Sod/Gpx/Cat系列基因的表达,并对检测结果进行半定量RT-PCR鉴定。结果Sod1/Sod2/Gpx1/Gpx3b/Gpx4b/Cat在各组小鼠骨髓细胞中均有较高的表达,Sod3/Gpx2则在7d组开始表达,11d组和14d组表达量持续减少。其中Sod2/Gpx3b/Gpx4b/Cat从0d组到14d组均为表达量先增加后减少;Sod1/Gpx1则为表达量先减少后增加;在各组小鼠骨髓细胞中未检测到Gpx3a/Gpx4a/Gpx5/Gpx6的表达。结论Sod/Gpx/Cat抗氧化酶系列基因在小鼠骨髓抑制和再生过程中呈现特征性的动态变化,这种变化提示了骨髓细胞氧化应激反应过程的组织特异性和调控特定的细胞信号通路的过程。
Objective To explore the dynamic expression profile of Sod/Gpx/Cat anti-oxidative genes during the depression and regeneration of mouse bone marrow, and the effect of oxidative stress on the regeneration of bone marrow.
Methods Models of bone marrow depression and regeneration were made by injecting 5-fluorouracil （5-Fu, 250mg/kg） into C57BL/6 mice, The mice were killed before and 3, 7, 11, and 14 days after the injection respectively （0 d, 3 d, 7 d, 11 d, and 14 d groups, 6 mice in each group） to collect the bone marrow-derived monocytes. Total RNA was extracted by using Trizol agent. And then the Sod/Gpx/Cat genes were detected using Affymetrix mouse cDNA 430A gene chips, and the results of detection were identified by semi-quantitative RT-PCR.
Results Sod1, Sod2, Gpx1, Gpx3b, Gpx4b, and Cat were all expressed at a high level in the bone marrow cells. Sod3 and Gpx2 began to express at 7 d group, and the expression level decreased continuously in 11 d group and 14 d group, the expression levels of Sod2, Gpx3b, Gpx4b, Cat increased at first and then gradually decreased; while the expression level of Sod1 and Gpx1 were decreased at first and then increased gradually. No Gpx3a, Gpx4a, Gpx5, or Gpx6 was detected in any of the groups.
Conclusions Sod/Gpx/Cat antioxidantive genes express dynamically during the depression and regeneration of mouse bone marrow, indicating that the oxidative stress, which is tissue-specified, can regulate the specific cell signal passway in the bone marrow cells.
出处
《中国医药生物技术》
CSCD
2008年第2期103-108,共6页
Chinese Medicinal Biotechnology
基金
国家自然科学基金(30570787
30572214)
国家高技术研究发展计划(863计划)(2007AA02Z149)
上海市科学技术委员会科研计划(05DZ19319)
