Splenectomy Modulates the Erythrocyte Turnover and Basigin (CD147) Expression in Mice
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ORIGINAL ARTICLE
Splenectomy Modulates the Erythrocyte Turnover and Basigin (CD147) Expression in Mice Nitin Bhardwaj1,2 • Ashutosh Singh3
Received: 3 October 2019 / Accepted: 26 February 2020 Ó Indian Society of Hematology and Blood Transfusion 2020
Abstract 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-H2DCFDA 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
& Nitin Bhardwaj [email protected]; [email protected] 1
Department of Zoology and Environmental Science, Gurukula Kangri Vishwavidyalaya, Haridwar, Uttarakhand 249404, India
2
School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India
3
Department of Biochemistry, University of Lucknow, Lucknow, UP 226007, India
expression may be a possible cause to increase erythrocyte removal in splenectomized mice. Keywords Erythrocyte Splenectomy Flow cytometry Double in vivo biotinylation Reactive oxygen species Basigin
Introduction Erythrocytes have a definite life span of 60 days in murine peripheral blood circulation. Approximately 2% erythrocytes are removed every day and are replaced with an equal number of fresh erythrocytes in mice [1]. Erythropoietic activity ensures that an equal number of new erythrocytes are added to blood circulation so that overall erythrocyte count in the blood remain constant [2]. During its survival erythrocytes travel hundreds of miles and are subjected to oxidant and osmotic stress with a limited repair mechanism to maintain itself. Under normal condition 80–90% of normal erythrocyte destruction activity is considered to be extravascular that takes place within tissue-resident macrophages of the spleen and to a lesser extent in the liver and bone marrow [3, 4]. Erythrocytes are phagocytosed at the rate of 2 9 106 per second by the macrophages. Each macrophage of spleen, liver and bone marrow phagocytose on ave
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