ROS及Nrf2相关抗氧化酶在红系发育中动态变化的研究  

Dynamic Changes of ROS and Nrf2-Related Factors During Erythroid Development

作　　者：杨蕾 吴凡 凌玲 郁多男 YANG Lei;WU Fan;LING Ling;YU Duo-Nan(Yangzhou University School of Medicine,Jiangsu Key Laboratory of Experimental&Translational Non-coding RNA Research,Yangzhou 225009,Jiangsu Province,China)

机构地区：[1]扬州大学医学院,江苏省非编码RNA基础与临床转化重点实验室,江苏扬州225009

出　　处：《中国实验血液学杂志》2023年第1期247-253,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(81670186,81870096);江苏省研究生科研与实践创新计划项目(KYCS22_3565)。

摘　　要：目的:研究活性氧自由基(ROS)及Nrf2相关因子在红系细胞发育过程中表达水平的变化和作用。方法:流式细胞术检测野生型小鼠外周血红细胞对强氧化剂过氧化氢的敏感性。通过流式分选得到不同发育阶段的骨髓红细胞并检测ROS水平。RT-q PCR方法检测骨髓红细胞各个发育阶段中Nrf2及相关抗氧化酶表达水平的变化。在Nrf2敲除小鼠中检测外周血和骨髓有核红细胞的ROS水平。体外实验检测野生型小鼠14.5 d胚胎肝细胞分选后的红系前体细胞在体外分化培养过程中Nrf2的表达水平。结果:野生型小鼠外周血中,过氧化氢处理后的网织红细胞ROS水平升高4倍左右,成熟红细胞ROS水平升高7倍左右(P<0.01)。在骨髓红细胞中,随着细胞发育成熟,ROS水平逐渐降低(r=0.85),而Nrf2(r=0.99)及相关抗氧化因子表达水平逐渐增高。与野生型小鼠相比,Nrf2敲除小鼠外周血红细胞和骨髓有核红细胞中的ROS水平均显著升高(P<0.01)。Nrf2在胚胎肝细胞分选后的红系细胞早期发育中表达升高(P<0.01)。结论:Nrf2及相关因子在红系细胞各分化阶段表达量不同,生理水平的Nrf2在红系细胞发育过程中发挥了重要的抗氧化作用。Objective:To investigate the changes and roles of reactive oxygen species (ROS) and nuclear factor erythroid 2-related factor 2 (Nrf2) related antioxidases during erythroid development.Methods:Flow cytometry was used to detect the sensibility of peripheral red blood cells of wild-type mice to a strong oxidant hydrogen peroxide (HO).Erythroid cells from different developmental stages in bone marrow (BM) were obtained using fluorescence-activ2at2ed cell sorter and the ROS levels were detected by flow cytometry.RT-q PCR was used to detect the changes of expression levels of Nrf2 and related antioxidases in erythroid cells from different developmental stages in BM.The ROS levels of the peripheral blood and BM nucleated erythrocytes in Nrf2 knockout mice were further examined.The expression level of Nrf2 in erythroid precursors isolated from 14.5 d embryonic liver of wild-type mice during differentiation and culture in vitro was detected.Results:In the peripheral blood of wild-type mice,the ROS level of reticulocytes and mature erythrocytes treated with H2O2increased about 4 times and 7 times,respectively (P<0.01).In BM erythrocytes,the ROS level gradually decreased as the cells matured (r=0.85),while the expression level of Nrf2 and its related anti-oxidative genes increased (r=0.99).The ROS levels in peripheral blood erythrocytes and BM nucleated erythrocytes of Nrf2knockout mice were significantly increased compared with wild-type mice (P<0.01).The expression of Nrf2 increased during the early erythroid development after embryonic liver cell sorting (P<0.01).Conclusion:The expression levels of Nrf2 and its related factors vary during erythropoiesis.Nrf2 at physiological level plays an important antioxidant role during the erythroid development.

关 键 词：红细胞 细胞发育 活性氧自由基 NRF2 

分 类 号：R555.1[医药卫生—血液循环系统疾病]