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.
Yangzhou University School of Medicine, Jiangsu Key Laboratory of Experimental & Translational Non-coding RNA Research, Yangzhou 225009, Jiangsu Province, China.E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):247-253. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.039.
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.
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-activated cell sorter and the ROS levels were detected by flow cytometry. RT-qPCR was used to detect the changes of expression levels of and related antioxidases in erythroid cells from different developmental stages in BM. The ROS levels of the peripheral blood and BM nucleated erythrocytes in knockout mice were further examined. The expression level of in erythroid precursors isolated from 14.5 d embryonic liver of wild-type mice during differentiation and culture in vitro was detected.
In the peripheral blood of wild-type mice, the ROS level of reticulocytes and mature erythrocytes treated with HO increased about 4 times and 7 times, respectively (<0.01). In BM erythrocytes, the ROS level gradually decreased as the cells matured (=0.85), while the expression level of and its related anti-oxidative genes increased (=0.99). The ROS levels in peripheral blood erythrocytes and BM nucleated erythrocytes of knockout mice were significantly increased compared with wild-type mice (<0.01). The expression of increased during the early erythroid development after embryonic liver cell sorting (<0.01).
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.
研究活性氧（ROS）和核因子红细胞2相关因子2（Nrf2）相关抗氧化酶在红细胞发育过程中的变化及作用。
采用流式细胞术检测野生型小鼠外周红细胞对强氧化剂过氧化氢（H₂O₂）的敏感性。利用荧光激活细胞分选仪获取骨髓（BM）中不同发育阶段的红细胞，并通过流式细胞术检测ROS水平。采用RT-qPCR检测BM中不同发育阶段红细胞中Nrf2及其相关抗氧化酶表达水平的变化。进一步检测Nrf2基因敲除小鼠外周血和BM有核红细胞的ROS水平。检测野生型小鼠14.5天胚胎肝脏分离的红系前体细胞在体外分化培养过程中Nrf2的表达水平。
在野生型小鼠外周血中，用H₂O₂处理的网织红细胞和成熟红细胞的ROS水平分别增加约4倍和7倍（P<0.01）。在BM红细胞中，随着细胞成熟，ROS水平逐渐降低（P=0.85），而Nrf2及其相关抗氧化基因的表达水平增加（P=0.99）。与野生型小鼠相比，Nrf2基因敲除小鼠外周血红细胞和BM有核红细胞的ROS水平显著升高（P<0.01）。胚胎肝细胞分选后早期红细胞发育过程中Nrf2的表达增加（P<0.01）。
Nrf2及其相关因子的表达水平在红细胞生成过程中有所不同。生理水平的Nrf2在红细胞发育过程中发挥重要的抗氧化作用。
