Bhardwaj Nitin, Singh Ashutosh
Department of Zoology and Environmental Science, Gurukula Kangri Vishwavidyalaya, Haridwar, Uttarakhand 249404 India.
School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067 India.
Indian J Hematol Blood Transfus. 2020 Oct;36(4):711-718. doi: 10.1007/s12288-020-01272-1. Epub 2020 Mar 3.
The present study was designed to study the splenectomy induced modulation of erythrocyte turnover in mice. We have also studied the modulation of reactive oxygen species (ROS) and basigin (CD147) expression level on erythrocytes in splenectomized condition. The erythrocyte turnover was studied by a newly developed double in vivo biotinylation (DIB) technique. This technique enables to discriminate three different age (young, intermediate and old) groups of erythrocytes. The expression level of ROS and CD147 was studied by staining with CM-HDCFDA stain and anti-mouse CD147 monocloclonal antibody followed by flow cytometry. We observed that intermediate and old age groups of erythrocytes were randomly eliminated in splenectomized condition. A marked surge in the blood reticulocyte count was observed in splenectomized mice. Splenectomy induced the level of ROS and CD147 expression on erythrocytes. The expression level of ROS was induced up to 35 days, but it reversed to basal level by 42 days indicating the emergence of refractoriness to splenectomy. The CD147 expression was significantly higher on day 7, 21 and 28 but it also normalizes on later time points. We conclude that erythrocyte turnover is significantly modulated in splenectomized mice. The enhanced level of ROS and CD147 expression may be a possible cause to increase erythrocyte removal in splenectomized mice.
本研究旨在探讨脾切除对小鼠红细胞更新的调节作用。我们还研究了脾切除状态下红细胞上活性氧(ROS)和基底膜蛋白(CD147)表达水平的调节情况。采用新开发的双体内生物素化(DIB)技术研究红细胞更新。该技术能够区分三种不同年龄(年轻、中年和老年)的红细胞群体。通过用CM-HDCFDA染料和抗小鼠CD147单克隆抗体染色,然后进行流式细胞术,研究ROS和CD147的表达水平。我们观察到,在脾切除状态下,中年和老年红细胞群体被随机清除。脾切除小鼠的血液网织红细胞计数显著增加。脾切除诱导了红细胞上ROS和CD147的表达水平。ROS的表达水平在35天内升高,但在42天时恢复到基础水平,表明对脾切除产生了耐受性。CD147的表达在第7天、21天和28天显著升高,但在随后的时间点也恢复正常。我们得出结论,脾切除小鼠的红细胞更新受到显著调节。ROS和CD147表达水平的升高可能是脾切除小鼠红细胞清除增加的一个可能原因。